LOCUS       DR274_CT_NT_mNG_        4816 bp    DNA        circular     02-AUG-2023
DEFINITION  .
ACCESSION   
VERSION     
SOURCE      .
  ORGANISM  .
COMMENT     Golden Gate reaction:
COMMENT     PCR pSap FRT unc-10  New Prime New Prime from 8 (BsaI) to 2938
            (BsaI) 
COMMENT     D274 CT-NT-bsaI II [3773] from 1707 (BsaI) to 1677 (BsaI) 
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     source          2..40
                     /locus_tag="Flex Linker"
                     /label="Flex Linker"
                     /ApEinfo_label="Flex Linker"
                     /ApEinfo_fwdcolor="#008080"
                     /ApEinfo_revcolor="#008080"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     primer_bind     complement(2920..2931)
                     /PCR_conditions="primer sequence:cggtctctgcgtgcctccggcact"
                     /locus_tag="New Primer"
                     /label="New Primer"
                     /ApEinfo_label="New Primer"
                     /ApEinfo_fwdcolor="#408000"
                     /ApEinfo_revcolor="#800080"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    join(41..146,198..326,378..508,560..784,836..949)
                     /locus_tag="NeonGreen_Cel"
                     /label="NeonGreen_Cel"
                     /ApEinfo_label="NeonGreen_Cel"
                     /ApEinfo_fwdcolor="#00ff00"
                     /ApEinfo_revcolor="#00ff00"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    509..559
                     /locus_tag="Synthetic Intron 3"
                     /label="Synthetic Intron 3"
                     /ApEinfo_label="Synthetic Intron 3"
                     /ApEinfo_fwdcolor="#ff80c0"
                     /ApEinfo_revcolor="#ff80c0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    147..197
                     /locus_tag="Synthetic Intron 1"
                     /label="Synthetic Intron 1"
                     /ApEinfo_label="Synthetic Intron 1"
                     /ApEinfo_fwdcolor="#ff80c0"
                     /ApEinfo_revcolor="#ff80c0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(1023..1039)
                     /PCR_conditions="primer
                     sequence:tggtctcagaccttagaggtcctcctcgg"
                     /locus_tag="New Primer(1)"
                     /label="New Primer(1)"
                     /ApEinfo_label="New Primer"
                     /ApEinfo_fwdcolor="#408000"
                     /ApEinfo_revcolor="#800080"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     intron          953..963
                     /locus_tag="intron"
                     /label="intron"
                     /ApEinfo_label="intron"
                     /ApEinfo_fwdcolor="#cc66ff"
                     /ApEinfo_revcolor="#ff00ff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1007..1036
                     /locus_tag="myc"
                     /label="myc"
                     /ApEinfo_label="myc"
                     /ApEinfo_fwdcolor="#6666ff"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     intron          998..1003
                     /locus_tag="intron(1)"
                     /label="intron(1)"
                     /ApEinfo_label="intron"
                     /ApEinfo_fwdcolor="#cc66ff"
                     /ApEinfo_revcolor="#ff00ff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2853..2886
                     /locus_tag="loxP"
                     /label="loxP"
                     /ApEinfo_label="loxP"
                     /ApEinfo_fwdcolor="#ff8000"
                     /ApEinfo_revcolor="#ff8000"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     intron          2887..2892
                     /locus_tag="intron(2)"
                     /label="intron(2)"
                     /ApEinfo_label="intron"
                     /ApEinfo_fwdcolor="#cc66ff"
                     /ApEinfo_revcolor="#ff00ff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1004..1006
                     /locus_tag="spacer"
                     /label="spacer"
                     /ApEinfo_label="spacer"
                     /ApEinfo_fwdcolor="#cccccc"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2893..2931
                     /locus_tag="flex linker"
                     /label="flex linker"
                     /ApEinfo_label="flex linker"
                     /ApEinfo_fwdcolor="#008080"
                     /ApEinfo_revcolor="#66ccff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     1..15
                     /PCR_conditions="primer sequence:tggtctctggcgggcagcggtgg"
                     /locus_tag="New Primer(2)"
                     /label="New Primer(2)"
                     /ApEinfo_label="New Primer"
                     /ApEinfo_fwdcolor="#408000"
                     /ApEinfo_revcolor="#800080"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    2..4
                     /locus_tag="GCG overlap"
                     /label="GCG overlap"
                     /ApEinfo_label="GCG overlap"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    785..835
                     /locus_tag="Synthetic Intron 4"
                     /label="Synthetic Intron 4"
                     /ApEinfo_label="Synthetic Intron 4"
                     /ApEinfo_fwdcolor="#ff80c0"
                     /ApEinfo_revcolor="#ff80c0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    327..377
                     /locus_tag="Synthetic Intron 2"
                     /label="Synthetic Intron 2"
                     /ApEinfo_label="Synthetic Intron 2"
                     /ApEinfo_fwdcolor="#ff80c0"
                     /ApEinfo_revcolor="#ff80c0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     1040..1066
                     /PCR_conditions="primer
                     sequence:tGGTCTCaGGTCcATTTTTGCTTTCGTCGTAAATC"
                     /locus_tag="New Primer(3)"
                     /label="New Primer(3)"
                     /ApEinfo_label="New Primer"
                     /ApEinfo_fwdcolor="#408000"
                     /ApEinfo_revcolor="#800080"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    950..952
                     /locus_tag="AAG overlap"
                     /label="AAG overlap"
                     /ApEinfo_label="AAG overlap"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             1450..2475
                     /note="hygromycinR"
                     /locus_tag="hygR"
                     /label="hygR"
                     /ApEinfo_label="hygR"
                     /ApEinfo_fwdcolor="#006666"
                     /ApEinfo_revcolor="#006666"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    964..997
                     /locus_tag="loxP(1)"
                     /label="loxP(1)"
                     /ApEinfo_label="loxP"
                     /ApEinfo_fwdcolor="#ff8000"
                     /ApEinfo_revcolor="#ff8000"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        complement(1044..1044)
                     /locus_tag="hsp16-41 promoter"
                     /label="hsp16-41 promoter"
                     /ApEinfo_label="hsp16-41 promoter"
                     /ApEinfo_fwdcolor="#0080c0"
                     /ApEinfo_revcolor="#0080c0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     3'UTR           2501..2852
                     /locus_tag="unc-54 3'UTR"
                     /label="unc-54 3'UTR"
                     /ApEinfo_label="unc-54 3'UTR"
                     /ApEinfo_fwdcolor="#990000"
                     /ApEinfo_revcolor="#cc0000"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        1045..1407
                     /note="Prps-0"
                     /note="/ApEinfo_fwdcolor=pink"
                     /note=""/ApEinfo_revcolor=pink; /ApEinfo_fwdcolor=pink;"
                     /locus_tag="Prps-0"
                     /label="Prps-0"
                     /ApEinfo_label="Prps-0"
                     /ApEinfo_fwdcolor="#339933"
                     /ApEinfo_revcolor="#339933"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(3117..3134)
                     /locus_tag="M13-fwd"
                     /label="M13-fwd"
                     /ApEinfo_label="M13-fwd"
                     /ApEinfo_fwdcolor="#808000"
                     /ApEinfo_revcolor="#800040"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     rep_origin      complement(3174..3577)
                     /locus_tag="New Feature"
                     /label="New Feature"
                     /ApEinfo_label="New Feature"
                     /ApEinfo_fwdcolor="#808080"
                     /ApEinfo_revcolor="#808080"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     rep_origin      complement(3578..3761)
                     /locus_tag="New Feature(1)"
                     /label="New Feature(1)"
                     /ApEinfo_label="New Feature"
                     /ApEinfo_fwdcolor="#808080"
                     /ApEinfo_revcolor="#808080"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             complement(3824..4639)
                     /locus_tag="KanR2"
                     /label="KanR2"
                     /ApEinfo_label="KanR2"
                     /ApEinfo_fwdcolor="#66ccff"
                     /ApEinfo_revcolor="#66ccff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     4718..4737
                     /locus_tag="NMoligo 5008"
                     /label="NMoligo 5008"
                     /ApEinfo_label="NMoligo 5008"
                     /ApEinfo_fwdcolor="#808000"
                     /ApEinfo_revcolor="#800040"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
ORIGIN
        1 gGCGGGCAGC GGTGGCAGTG GAGGTACCGG CGGAAGCGGT ATGGTGTCGA AGGGAGAAGA
       61 GGATAACATG GCTTCACTCC CAGCTACACA CGAACTCCAC ATCTTCGGAT CGATCAACGG
      121 AGTGGATTTC GATATGGTCG GACAAGgtaa gtttaaacat atatatacta actaaccctg
      181 attatttaaa ttttcagGAA CTGGAAACCC AAACGATGGA TACGAGGAAC TCAACCTCAA
      241 GTCGACAAAG GGAGATCTGC AATTCTCGCC ATGGATTCTC GTGCCACACA TCGGATACGG
      301 ATTCCACCAA TACCTCCCAT ACCCAGgtaa gtttaaactg agttctacta actaacgagt
      361 aatatttaaa ttttcagATG GAATGTCACC ATTCCAAGCT GCCATGGTGG ATGGATCGGG
      421 ATACCAAGTT CACCGAACAA TGCAATTCGA GGATGGAGCC TCGCTCACAG TGAACTACCG
      481 ATACACATAC GAGGGATCGC ACATCAAGgt aagtttaaac agttcggtac taactaacca
      541 tacatattta aattttcagG GAGAGGCTCA AGTTAAGGGA ACAGGATTCC CAGCTGATGG
      601 ACCAGTGATG ACAAACTCAC TCACAGCTGC TGATTGGTGC CGATCGAAAA AGACATACCC
      661 AAATGATAAG ACAATCATCT CGACATTCAA GTGGTCGTAC ACTACTGGAA ACGGAAAGCG
      721 ATACCGATCG ACAGCCCGAA CAACATACAC ATTCGCTAAG CCAATGGCCG CCAACTACCT
      781 CAAGgtaagt ttaaacatga ttttactaac taactaatct gatttaaatt ttcagAATCA
      841 ACCAATGTAC GTGTTCCGAA AGACAGAACT CAAGCACTCA AAGACAGAGC TGAACTTCAA
      901 AGAGTGGCAA AAGGCCTTCA CAGATGTGAT GGGAATGGAT GAACTCTACA AGgtaagttt
      961 aaaATAACTT CGTATAGCAT ACATTATACG AAGTTATttt cagGGAGAGC AAAAGCTTAT
     1021 CTCCGAGGAG GACCTCTAAG GTCcATTTTT GCTTTCGTCG TAAATCTACA CACGCGTCTC
     1081 TTCCGTGCGA GAGTCCAAGC CAGCAGCCAA ATTCGTTGAC TGAGTATTCA ACGTTTATAC
     1141 GTTGTCGGCA ACGAGAAATA GGAAAATGCA TCGGGAAATG TTCTTTTTTC GATTTTTTCC
     1201 AAGGTTTTGA CAAATTTTAC CACGAATTTT GCTATGTTTT CAATTAAAAA ATATGTTATT
     1261 CAACTGTTTC TATGAGGAAA ATAAGGCTTT GCATGTAATT TTCTTATTCA GCATAATTTT
     1321 TAATTAATTT GAATTTTCTG TCCTAACGTT TATTTTGTTT TCTTGGTTAT GACTGATCTG
     1381 AAATTAATTT TTGAATTTTA AGGTAATATG TCAGGCGGTG CCGCAAGTTT GTACAAAAAA
     1441 GCAGGCTCCA TGAAAAAGCC TGAACTCACC GCGACGTCTG TCGAGAAGTT TCTGATCGAA
     1501 AAGTTCGACA GCGTCTCCGA CCTGATGCAG CTCTCGGAGG GCGAAGAATC TCGTGCTTTC
     1561 AGCTTCGATG TAGGAGGGCG TGGATATGTC CTGCGGGTAA ATAGCTGCGC CGATGGTTTC
     1621 TACAAAGATC GTTATGTTTA TCGGCACTTT GCATCGGCCG CGCTCCCGAT TCCGGAAGTG
     1681 CTTGACATTG GGGAATTCAG CGAGAGCCTG ACCTATTGCA TCTCCCGCCG TGCACAGGGT
     1741 GTCACGTTGC AAGACCTGCC TGAAACCGAA CTGCCCGCTG TTCTGCAGCC GGTCGCGGAG
     1801 GCCATGGATG CGATCGCTGC GGCCGATCTT AGCCAGACGA GCGGGTTCGG CCCATTCGGA
     1861 CCGCAAGGAA TCGGTCAATA CACTACATGG CGTGATTTCA TATGCGCGAT TGCTGATCCC
     1921 CATGTGTATC ACTGGCAAAC TGTGATGGAC GACACCGTCA GTGCGTCCGT CGCGCAGGCT
     1981 CTCGATGAGC TGATGCTTTG GGCCGAGGAC TGCCCCGAAG TCCGGCACCT CGTGCACGCG
     2041 GATTTCGGCT CCAACAATGT CCTGACGGAC AATGGCCGCA TAACAGCGGT CATTGACTGG
     2101 AGCGAGGCGA TGTTCGGGGA TTCCCAATAC GAGGTCGCCA ACATCTTCTT CTGGAGGCCG
     2161 TGGTTGGCTT GTATGGAGCA GCAGACGCGC TACTTCGAGC GGAGGCATCC GGAGCTTGCA
     2221 GGATCGCCGC GGCTCCGGGC GTATATGCTC CGCATTGGTC TTGACCAACT CTATCAGAGC
     2281 TTGGTTGACG GCAATTTCGA TGATGCAGCT TGGGCGCAGG GTCGATGCGA CGCAATCGTC
     2341 CGATCCGGAG CCGGGACTGT CGGGCGTACA CAAATCGCCC GCAGAAGCGC GGCCGTCTGG
     2401 ACCGATGGCT GTGTAGAAGT ACTCGCCGAT AGTGGAAACC GACGCCCCAG CACTCGTCCG
     2461 AGGGCAAAGG AATAGACCCA GCTTTCTTGT ACAAAGTGGG TCCAATTACT CTTCAACATC
     2521 CCTACATGCT CTTTCTCCCT GTGCTCCCAC CCCCTATTTT TGTTATTATC AAAAAACTTC
     2581 TCTTAATTTC TTTGTTTTTT AGCTTCTTTT AAGTCACCTC TAACAATGAA ATTGTGTAGA
     2641 TTCAAAAATA GAATTAATTC GTAATAAAAA GTCGAAAAAA ATTGTGCTCC CTCCCCCCAT
     2701 TAATAATAAT TCTATCCCAA AATCTACACA ATGTTCTGTG TACACTTCTT ATGTTTTTTA
     2761 CTTCTGATAA ATTTTTTTGA AACATCATAG AAAAAACCGC ACACAAAATA CCTTATCATA
     2821 TGTTACGTTT CAGTTTATGA CCGCAATTTT TAATAACTTC GTATAGCATA CATTATACGA
     2881 AGTTATtttc agGGTGGCAG CGCTGGAGGT ACCGGCGGTA GTGCCGGAGG CACGcGAAGA
     2941 GCAAGCTTGG ATCGACGAGA GCAGCGCGAC TGGATCTGTC GCCCGTCTCA AACGCAACCC
     3001 TCCGGCGGTC GCATATCATT CAGGACGAGC CTCAGACTCC AGCGTAACTG GACTGCAATC
     3061 AACTCACTGG CTCACCTTCC GGTCCACGAT CAGCTAGAAT CAAGCTGACT AGATAAACTG
     3121 GCCGTCGTTT TACACGGGTG GGCCTTTCTT CGGTAGAAAA TCAAAGGATC TTCttgagat
     3181 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3241 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag
     3301 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3361 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3421 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3481 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3541 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3601 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3661 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc
     3721 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aAACGCCAGC AACGCAGAAA
     3781 GGCCCACCCG AAGGTGAGCC AGGTGATTAC ATTTAGGTCC TCATTAGAAA AACTCATCGA
     3841 GCATCAAGTG AAACTGCAAT TTATTCATAT CAGGATTATC AATACCATAT TTTTGAAAAA
     3901 GCCGTTTCTG TAATGAAGGA GAAAACTCAC CGAGGCAGTT CCATAGGATG GCAAGATCCT
     3961 GGTATCGGTC TGCGATTCCG ACTCGTCCAA CATCAATACA ACCTATTAAT TTCCCCTCGT
     4021 CAAAAATAAG GTTATCAAGT GAGAAATCAC CATGAGTGAC GACTGAATCC GGTGAGAATG
     4081 GCAAGAGTTT ATGCATTTCT TTCCAGACTT GTTCAACAGG CCAGCCATTA CGCTCGTCAT
     4141 CAAAATCACT CGCACCAACC AAACCGTTAT TCATTCGTGA TTGCGCCTGA GCGAGACGAA
     4201 ATACGCGATC GCCGTTAAAA GGACAATTAC AAACAGGAAT CGAATGCAAC CGGCGCAGGA
     4261 ACACTGCCAG CGCATCAACA ATATTTTCAC CTGAATCAGG ATATTCTTCT AATACCTGGA
     4321 ATGCTGTTTT CCCTGGGATC GCAGTGGTGA GTAACCATGC ATCATCAGGA GTACGGATAA
     4381 AATGCTTGAT GGTCGGAAGA GGCATAAATT CCGTCAGCCA GTTTAGCCTG ACCATCTCAT
     4441 CTGTAACATC ATTGGCAACG CTACCTTTGC CATGTTTCAG AAACAACTCT GGCGCATCGG
     4501 GCTTCCCATA CAATCGATAG ATTGTCGCAC CTGATTGCCC GACATTATCG CGAGCCCATT
     4561 TATACCCATA TAAATCAGCA TCCATGTTGG AATTTAATCG CGGCTTCAAG CAAGACGTTT
     4621 CCCGTTGAAT ATGGCTCATT TTAGCTTCCT TAGCTCCTGA AAATCTCGAT AACTCAAAAA
     4681 ATACGCCCGG TAGTGATCTT ATTTCATTAT GGTGAAAgtt ggaacctctt acgtgccGAT
     4741 CAAGTCAAAA GCCTCCGGTC GGAGGCTTTT GACTTTCTGC TATGGAGGTC AGGTATGATT
     4801 TttGAATTCG CTCTTC    
//
