LOCUS       pNeoG1r                 5797 bp    DNA        circular     19-MAR-2022
DEFINITION  .
ACCESSION   
VERSION     
SOURCE      .
  ORGANISM  .
COMMENT     Golden Gate reaction:
COMMENT     pHygG minus hyg PCR from 386 (BsaI) to 362 (BsaI) 
COMMENT     NEo PCR 1 from 8 (BsaI) to 485 (BsaI) 
COMMENT     neoPCR2 from 8 (BsaI) to 215 (BsaI) 
COMMENT     neoPCR3 from 8 (BsaI) to 114 (BsaI) 
COMMENT     
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     promoter        complement(5435..5796)
                     /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"
     misc_feature    complement(4642..5433)
                     /locus_tag="NeoR"
                     /label="NeoR"
                     /ApEinfo_label="NeoR"
                     /ApEinfo_fwdcolor="#7980ff"
                     /ApEinfo_revcolor="#7980ff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 1 offset 0"
     CDS             complement(1033..1692)
                     /locus_tag="AmpR"
                     /label="AmpR"
                     /ApEinfo_label="AmpR"
                     /ApEinfo_fwdcolor="#ff8000"
                     /ApEinfo_revcolor="#ff8000"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    3758..4159
                     /locus_tag="myo-2 3' UTR"
                     /label="myo-2 3' UTR"
                     /ApEinfo_label="myo-2 3' UTR"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2747..2749
                     /locus_tag="pMYO-2"
                     /label="pMYO-2"
                     /ApEinfo_label="pMYO-2"
                     /ApEinfo_fwdcolor="#85dae9"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2710..2743
                     /locus_tag="FRT"
                     /label="FRT"
                     /ApEinfo_label="FRT"
                     /ApEinfo_fwdcolor="#800000"
                     /ApEinfo_revcolor="#008080"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2886..2936
                     /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"
     misc_feature    2515..2548
                     /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"
     misc_feature    3066..3116
                     /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"
     rep_origin      complement(2153..2459)
                     /locus_tag="F1 ori"
                     /label="F1 ori"
                     /ApEinfo_label="F1 ori"
                     /ApEinfo_fwdcolor="#808080"
                     /ApEinfo_revcolor="#808080"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    join(2780..2885,2937..3065,3117..3247,3299..3523,
                     3575..3694)
                     /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"
     transit_peptide 3695..3751
                     /locus_tag="egl-13 NLS"
                     /label="egl-13 NLS"
                     /ApEinfo_label="egl-13 NLS"
                     /ApEinfo_fwdcolor="#ff0080"
                     /ApEinfo_revcolor="#ff0080"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     rep_origin      complement(248..935)
                     /locus_tag="ColE1 origin"
                     /label="ColE1 origin"
                     /ApEinfo_label="ColE1 origin"
                     /ApEinfo_fwdcolor="#808080"
                     /ApEinfo_revcolor="#808080"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    3524..3574
                     /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"
     3'UTR           complement(4286..4637)
                     /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"
     sig_peptide     2747..2779
                     /locus_tag="SV40 nls"
                     /label="SV40 nls"
                     /ApEinfo_label="SV40 nls"
                     /ApEinfo_fwdcolor="#ff0080"
                     /ApEinfo_revcolor="#ff0080"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    3248..3298
                     /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    complement(2656..2689)
                     /locus_tag="FRT3"
                     /label="FRT3"
                     /ApEinfo_label="FRT3"
                     /ApEinfo_fwdcolor="#804000"
                     /ApEinfo_revcolor="#008040"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2591..2656
                     /locus_tag="MCS"
                     /label="MCS"
                     /ApEinfo_label="MCS"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
ORIGIN
        1 atcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
       61 cggggagagg cggtttgcgt attgggcgaa ttccgcttcc tcgctcactg actcgctgcg
      121 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
      181 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
      241 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      301 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
      361 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
      421 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
      481 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
      541 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
      601 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
      661 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
      721 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
      781 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
      841 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
      901 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
      961 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1021 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     1081 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     1141 atctggcccc agtgctgcaa tgataccgcg agatccacgc tcaccggctc cagatttatc
     1201 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     1261 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     1321 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     1381 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     1441 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     1501 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     1561 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     1621 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     1681 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     1741 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     1801 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     1861 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     1921 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     1981 aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt
     2041 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     2101 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     2161 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     2221 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     2281 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     2341 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     2401 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     2461 ccgctacagg gcgcgtcgtc ggagctccac cgcggtggcg gccgctCTAG ATGGATAACT
     2521 TCGTATAGCA TACATTATAC GAAGTTATAC Gggcgaattg gagctccacc gcggtggcgg
     2581 ccgctctaga ACTAGTGGTA CCGAAGAGCa tGCaaCGCGT ggttaccTGC AGGCTCTTCG
     2641 CCACTTAAGA cccggGAAGT TCCTATACta tttgaaGAAT AGGAACTTCt gcaggaattc
     2701 gatatcGGTG AAGTTCCTAT TCtctagaaa GTATAGGAAC TTCAAAATGG GATCCCCAAA
     2761 GAAGAAGCGT AAGGTGTCGA TGGTGTCGAA GGGAGAAGAG GATAACATGG CTTCACTCCC
     2821 AGCTACACAC GAACTCCACA TCTTCGGATC GATCAACGGA GTGGATTTCG ATATGGTCGG
     2881 ACAAGgtaag tttaaacata tatatactaa ctaaccctga ttatttaaat tttcagGAAC
     2941 TGGAAACCCA AACGATGGAT ACGAGGAACT CAACCTCAAG TCGACAAAGG GAGATCTGCA
     3001 ATTCTCGCCA TGGATTCTCG TGCCACACAT CGGATACGGA TTCCACCAAT ACCTCCCATA
     3061 CCCAGgtaag tttaaactga gttctactaa ctaacgagta atatttaaat tttcagATGG
     3121 AATGTCACCA TTCCAAGCTG CCATGGTGGA TGGATCGGGA TACCAAGTTC ACCGAACAAT
     3181 GCAATTCGAG GATGGAGCCT CGCTCACAGT GAACTACCGA TACACATACG AGGGATCGCA
     3241 CATCAAGgta agtttaaaca gttcggtact aactaaccat acatatttaa attttcagGG
     3301 AGAGGCTCAA GTTAAGGGAA CAGGATTCCC AGCTGATGGA CCAGTGATGA CAAACTCACT
     3361 CACAGCTGCT GATTGGTGCC GATCGAAAAA GACATACCCA AATGATAAGA CAATCATCTC
     3421 GACATTCAAG TGGTCGTACA CTACTGGAAA CGGAAAGCGA TACCGATCGA CAGCCCGAAC
     3481 AACATACACA TTCGCTAAGC CAATGGCCGC CAACTACCTC AAGgtaagtt taaacatgat
     3541 tttactaact aactaatctg atttaaattt tcagAATCAA CCAATGTACG TGTTCCGAAA
     3601 GACAGAACTC AAGCACTCAA AGACAGAGCT GAACTTCAAA GAGTGGCAAA AGGCCTTCAC
     3661 AGATGTGATG GGAATGGATG AACTCTACAA GGCCTCCCGT CGTCGTAAGG CCAACCCAAC
     3721 CAAGCTCTCC GAGAACGCCA AGAAGCTCGC CAAGtagatc aactgctcta attccatctc
     3781 gtttctctct tttctttcaa tctctttacc cgtttctctt cccctacgtc ccccaaacct
     3841 accactgcac agtaacacat acacattcaa cgtgtcacaa taaaagccca tcaacacctt
     3901 ctctgttgtt ttttttcttc aaatgcattt ttttcacttt gtttaataat ttttacgcat
     3961 cggatttgta ggatagtgga aatttgtcct tcaatgcttc aattgattcg acacttctca
     4021 atatttttga tcaataaacc attgattttc tatttagaag aagatcactg taaataacac
     4081 caaaaacatc acagcaaaaa aactcccact caattatatt actgttttta ttgaaactag
     4141 aataaatttg aaaatcttca aagcatacac aattaaaaaa aacgaaaaaa gaaaaagtcc
     4201 cgttgattct caaacttaaa tttttaaact tgtctgaacc atgttgtaag tttgtaaaag
     4261 tttcccagtt gtcctcgaac aGGTCTAAAA ATTGCGGTCA TAAACTGAAA CGTAACATAT
     4321 GATAAGGTAT TTTGTGTGCG GTTTTTTCTA TGATGTTTCA AAAAAATTTA TCAGAAGTAA
     4381 AAAACATAAG AAGTGTACAC AGAACATTGT GTAGATTTTG GGATAGAATT ATTATTAATG
     4441 GGGGGAGGGA GCACAATTTT TTTCGACTTT TTATTACGAA TTAATTCTAT TTTTGAATCT
     4501 ACACAATTTC ATTGTTAGAG GTGACTTAAA AGAAGCTAAA AAACAAAGAA ATTAAGAGAA
     4561 GTTTTTTGAT AATAACAAAA ATAGGGGGTG GGAGCACAGG GAGAAAGAGC ATGTAGGGAT
     4621 GTTGAAGAGT AATTGGACTC AGAAGAACTC GTCAAGAAGG CGATAGAAGG CGATGCGCTG
     4681 CGAATCGGGA GCGGCGATAC CGTAAAGCAC GAGGAAGCGG TCAGCCCATT CGCCGCCAAG
     4741 CTCCTCAGCA ATATCACGGG TAGCCAACGC TATGTCCTGA TAGCGGTCCG CCACACCCAG
     4801 CCGGCCACAG TCGATGAATC CAGAAAAGCG GCCATTTTCC ACCATGATAT TCGGCAAGCA
     4861 GGCATCGCCA TGGGTCACGA CGAGATCCTC GCCGTCGGGC ATGCGCGCCT TGAGCCTGGC
     4921 GAACAGTTCG GCTGGTGCGA GCCCCTGATG TTCTTCGTCC AGATCATCCT GATCGACAAG
     4981 ACCGGCTTCC ATCCGAGTAC GTGCTCGCTC GATGCGATGT TTCGCTTGGT GGTCGAATGG
     5041 GCAGGTAGCC GGATCAAGCG TATGCAGCCG CCGCATTGCA TCAGCCATGA TGGATACTTT
     5101 CTCGGCAGGA GCAAGGTGAG ATGACAGGAG ATCCTGCCCC GGCACTTCGC CCAATAGCAG
     5161 CCAGTCCCTT CCCGCTTCAG TGACAACGTC GAGCACAGCT GCGCAAGGAA CGCCCGTCGT
     5221 GGCCAGCCAC GATAGCCGCG CTGCCTCGTC CTGCAGTTCA TTCAGGGCAC CGGACAGGTC
     5281 GGTCTTGACA AAAAGAACCG GGCGCCCCTG CGCTGACAGC CGGAACACGG CGGCATCAGA
     5341 GCAGCCGATT GTCTGTTGTG CCCAGTCATA GCCGAATAGC CTCTCCACCC AAGCGGCCGG
     5401 AGAACCTGCG TGCAATCCAT CTTGTTCAAT CATATTACCT TAAAATTCAA AAATTAATTT
     5461 CAGATCAGTC ATAACCAAGA AAACAAAATA AACGTTAGGA CAGAAAATTC AAATTAATTA
     5521 AAAATTATGC TGAATAAGAA AATTACATGC AAAGCCTTAT TTTCCTCATA GAAACAGTTG
     5581 AATAACATAT TTTTTAATTG AAAACATAGC AAAATTCGTG GTAAAATTTG TCAAAACCTT
     5641 GGAAAAAATC GAAAAAAGAA CATTTCCCGA TGCATTTTCC TATTTCTCGT TGCCGACAAC
     5701 GTATAAACGT TGAATACTCA GTCAACGAAT TTGGCTGCTG GCTTGGACTC TCGCACGGAA
     5761 GAGACGCGTG TGTAGATTTA CGACGAAAGC AAAAATg  
//
