LOCUS       pHygCG3                 5753 bp    DNA        circular     04-JUN-2022
DEFINITION  .
ACCESSION   
VERSION     
SOURCE      .
  ORGANISM  .
COMMENT     Golden Gate reaction:
COMMENT     pHygcG3 from 8 (BsaI) to 4519 (BsaI) 
COMMENT     mNG tbb-2 PCR from 8 (BsaI) to 1248 (BsaI) 
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     CDS             complement(786..1445)
                     /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    complement(2344..2377)
                     /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    join(2435..2540,2592..2720,2772..2902,2954..3178,
                     3230..3346)
                     /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    2290..2344
                     /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"
     misc_feature    2721..2771
                     /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(1906..2212)
                     /locus_tag="F1 ori"
                     /label="F1 ori"
                     /ApEinfo_label="F1 ori"
                     /ApEinfo_fwdcolor="#808080"
                     /ApEinfo_revcolor="#808080"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_recomb     2398..2431
                     /locus_tag="FRT"
                     /label="FRT"
                     /ApEinfo_label="FRT"
                     /ApEinfo_fwdcolor="#800000"
                     /ApEinfo_revcolor="#800000"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    2241..2274
                     /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"
     3'UTR           5155..5506
                     /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"
     misc_feature    3179..3229
                     /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"
     promoter        3699..4061
                     /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"
     CDS             4104..5129
                     /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_recomb     complement(5133..5153)
                     /locus_tag="attB2"
                     /label="attB2"
                     /ApEinfo_label="attB2"
                     /ApEinfo_fwdcolor="#dda0dd"
                     /ApEinfo_revcolor="#dda0dd"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_recomb     4078..4098
                     /locus_tag="attB1"
                     /label="attB1"
                     /ApEinfo_label="attB1"
                     /ApEinfo_fwdcolor="#dda0dd"
                     /ApEinfo_revcolor="#dda0dd"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    complement(5140..5146)
                     /locus_tag="Gateway Recomb2"
                     /label="Gateway Recomb2"
                     /ApEinfo_label="Gateway Recomb2"
                     /ApEinfo_fwdcolor="#ff0000"
                     /ApEinfo_revcolor="#ff0000"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    4085..4091
                     /locus_tag="Gateway recomb1"
                     /label="Gateway recomb1"
                     /ApEinfo_label="Gateway recomb1"
                     /ApEinfo_fwdcolor="#ff0000"
                     /ApEinfo_revcolor="#ff0000"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     rep_origin      complement(1..688)
                     /locus_tag="ColE1 origin"
                     /label="ColE1 origin"
                     /ApEinfo_label="ColE1 origin"
                     /ApEinfo_fwdcolor="#808080"
                     /ApEinfo_revcolor="#808080"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    2903..2953
                     /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    2541..2591
                     /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"
     3'UTR           3350..3675
                     /locus_tag="tbb-2 3'UTR"
                     /label="tbb-2 3'UTR"
                     /ApEinfo_label="tbb-2 3'UTR"
                     /ApEinfo_fwdcolor="#cc66ff"
                     /ApEinfo_revcolor="#cc66ff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
ORIGIN
        1 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
       61 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      121 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
      181 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
      241 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cattcagccc
      301 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
      361 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
      421 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
      481 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
      541 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
      601 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
      661 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
      721 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
      781 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
      841 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
      901 cccagtgctg caatgatacc gcgagatcca cgctcaccgg ctccagattt atcagcaata
      961 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     1021 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     1081 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     1141 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     1201 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     1261 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     1321 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     1381 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     1441 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     1501 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     1561 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     1621 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     1681 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     1741 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     1801 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     1861 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     1921 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     1981 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     2041 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     2101 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     2161 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     2221 agggcgcgtc gtcggagctc ATAACTTCGT ATAGCATACA TTATACGAAG TTATgcggcc
     2281 gctctagaaC TAGTGGCGAA GAGCCTGCAg gtaaccACGC GttGCatGCT CTTCGGTAcc
     2341 cggGAAGTTC CTATACtatt tgaaGAATAG GAACTTCtgc aggaattcga tatcGGTGAA
     2401 GTTCCTATTC tctagaaaGT ATAGGAACTT CAAAATGGTG TCGAAGGGAG AAGAGGATAA
     2461 CATGGCTTCA CTCCCAGCTA CACACGAACT CCACATCTTC GGATCGATCA ACGGAGTGGA
     2521 TTTCGATATG GTCGGACAAG gtaagtttaa acatatatat actaactaac cctgattatt
     2581 taaattttca gGAACTGGAA ACCCAAACGA TGGATACGAG GAACTCAACC TCAAGTCGAC
     2641 AAAGGGAGAT CTGCAATTCT CGCCATGGAT TCTCGTGCCA CACATCGGAT ACGGATTCCA
     2701 CCAATACCTC CCATACCCAG gtaagtttaa actgagttct actaactaac gagtaatatt
     2761 taaattttca gATGGAATGT CACCATTCCA AGCTGCCATG GTGGATGGAT CGGGATACCA
     2821 AGTTCACCGA ACAATGCAAT TCGAGGATGG AGCCTCGCTC ACAGTGAACT ACCGATACAC
     2881 ATACGAGGGA TCGCACATCA AGgtaagttt aaacagttcg gtactaacta accatacata
     2941 tttaaatttt cagGGAGAGG CTCAAGTTAA GGGAACAGGA TTCCCAGCTG ATGGACCAGT
     3001 GATGACAAAC TCACTCACAG CTGCTGATTG GTGCCGATCG AAAAAGACAT ACCCAAATGA
     3061 TAAGACAATC ATCTCGACAT TCAAGTGGTC GTACACTACT GGAAACGGAA AGCGATACCG
     3121 ATCGACAGCC CGAACAACAT ACACATTCGC TAAGCCAATG GCCGCCAACT ACCTCAAGgt
     3181 aagtttaaac atgattttac taactaacta atctgattta aattttcagA ATCAACCAAT
     3241 GTACGTGTTC CGAAAGACAG AACTCAAGCA CTCAAAGACA GAGCTGAACT TCAAAGAGTG
     3301 GCAAAAGGCC TTCACAGATG TGATGGGAAT GGATGAACTC TACAAGACGt aaatgcaaaa
     3361 tcctttcaag cattcccttc ttctctatca ctcttctttc tttttgtcaa aaaattctct
     3421 cgctaattta tttgcttttt taatgttatt attttatgac tttttatagt cactgaaaag
     3481 tttgcatctg agtgaagtga atgctatcaa aatgtgattc tgtctgatgt actttcacaa
     3541 tctctcttca attccatttt gaagtgcttt aaacccgaaa ggttgagaaa aatgcgagcg
     3601 ctcaaatatt tgtattgtgt tcgttgagtg acccaacaaa aagaggaaac tttattgtgc
     3661 cgccaagaaa aaagtctcag ttgtcctcga acaGGTCcAT TTTTGCTTTC GTCGTAAATC
     3721 TACACACGCG TCTCTTCCGT GCGAGAGTCC AAGCCAGCAG CCAAATTCGT TGACTGAGTA
     3781 TTCAACGTTT ATACGTTGTC GGCAACGAGA AATAGGAAAA TGCATCGGGA AATGTTCTTT
     3841 TTTCGATTTT TTCCAAGGTT TTGACAAATT TTACCACGAA TTTTGCTATG TTTTCAATTA
     3901 AAAAATATGT TATTCAACTG TTTCTATGAG GAAAATAAGG CTTTGCATGT AATTTTCTTA
     3961 TTCAGCATAA TTTTTAATTA ATTTGAATTT TCTGTCCTAA CGTTTATTTT GTTTTCTTGG
     4021 TTATGACTGA TCTGAAATTA ATTTTTGAAT TTTAAGGTAA TATGTCAGGC GGTGCCGCAA
     4081 GTTTGTACAA AAAAGCAGGC TCCATGAAAA AGCCTGAACT CACCGCGACG TCTGTCGAGA
     4141 AGTTTCTGAT CGAAAAGTTC GACAGCGTCT CCGACCTGAT GCAGCTCTCG GAGGGCGAAG
     4201 AATCTCGTGC TTTCAGCTTC GATGTAGGAG GGCGTGGATA TGTCCTGCGG GTAAATAGCT
     4261 GCGCCGATGG TTTCTACAAA GATCGTTATG TTTATCGGCA CTTTGCATCG GCCGCGCTCC
     4321 CGATTCCGGA AGTGCTTGAC ATTGGGGAAT TCAGCGAGAG CCTGACCTAT TGCATCTCCC
     4381 GCCGTGCACA GGGTGTCACG TTGCAAGACC TGCCTGAAAC CGAACTGCCC GCTGTTCTGC
     4441 AGCCGGTCGC GGAGGCCATG GATGCGATCG CTGCGGCCGA TCTTAGCCAG ACGAGCGGGT
     4501 TCGGCCCATT CGGACCGCAA GGAATCGGTC AATACACTAC ATGGCGTGAT TTCATATGCG
     4561 CGATTGCTGA TCCCCATGTG TATCACTGGC AAACTGTGAT GGACGACACC GTCAGTGCGT
     4621 CCGTCGCGCA GGCTCTCGAT GAGCTGATGC TTTGGGCCGA GGACTGCCCC GAAGTCCGGC
     4681 ACCTCGTGCA CGCGGATTTC GGCTCCAACA ATGTCCTGAC GGACAATGGC CGCATAACAG
     4741 CGGTCATTGA CTGGAGCGAG GCGATGTTCG GGGATTCCCA ATACGAGGTC GCCAACATCT
     4801 TCTTCTGGAG GCCGTGGTTG GCTTGTATGG AGCAGCAGAC GCGCTACTTC GAGCGGAGGC
     4861 ATCCGGAGCT TGCAGGATCG CCGCGGCTCC GGGCGTATAT GCTCCGCATT GGTCTTGACC
     4921 AACTCTATCA GAGCTTGGTT GACGGCAATT TCGATGATGC AGCTTGGGCG CAGGGTCGAT
     4981 GCGACGCAAT CGTCCGATCC GGAGCCGGGA CTGTCGGGCG TACACAAATC GCCCGCAGAA
     5041 GCGCGGCCGT CTGGACCGAT GGCTGTGTAG AAGTACTCGC CGATAGTGGA AACCGACGCC
     5101 CCAGCACTCG TCCGAGGGCA AAGGAATAGA CCCAGCTTTC TTGTACAAAG TGGGTCCAAT
     5161 TACTCTTCAA CATCCCTACA TGCTCTTTCT CCCTGTGCTC CCACCCCCTA TTTTTGTTAT
     5221 TATCAAAAAA CTTCTCTTAA TTTCTTTGTT TTTTAGCTTC TTTTAAGTCA CCTCTAACAA
     5281 TGAAATTGTG TAGATTCAAA AATAGAATTA ATTCGTAATA AAAAGTCGAA AAAAATTGTG
     5341 CTCCCTCCCC CCATTAATAA TAATTCTATC CCAAAATCTA CACAATGTTC TGTGTACACT
     5401 TCTTATGTTT TTTACTTCTG ATAAATTTTT TTGAAACATC ATAGAAAAAA CCGCACACAA
     5461 AATACCTTAT CATATGTTAC GTTTCAGTTT ATGACCGCAA TTTTTAatcc cgctttccag
     5521 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5581 ttgcgtattg ggcgaattcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5641 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5701 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgt
//
