LOCUS       62056_12700        6621 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6621)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1891..1907
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2127..2834
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2882..2898)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2906..2922)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(2930..2960)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2975..2996)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3284..3872)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             4076..4876
                     /codon_start=1
                     /label=ApmR
                     /note="aminoglycoside 3-N-acetyltransferase type IV"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     CDS             complement(5313..5681)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(5714..5823)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(6061..6072)
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     protein_bind    6143..6242
                     /label=phage phi-C31 attP
                     /note="attachment site of phage phi-C31"
     CDS             join(6252..6621,1..1445)
                     /codon_start=1
                     /label=phage phi-C31 integrase
                     /note="integrase from phage phi-C31"
                     /translation="VDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
ORIGIN
        1 tccggcaggg aaacgtcatg gacctgattc acctgattat gcggctcgac gcgtcgcaca
       61 aagaatcttc gctgaagtcg gcgaagattc tcgacacgaa gaaccttcag cgcgaattgg
      121 gcgggtacgt cggcgggaag gcgccttacg gcttcgagct tgtttcggag acgaaggaga
      181 tcacgcgcaa cggccgaatg gtcaatgtcg tcatcaacaa gcttgcgcac tcgaccactc
      241 cccttaccgg acccttcgag ttcgagcccg acgtaatccg gtggtggtgg cgtgagatca
      301 agacgcacaa acaccttccc ttcaagccgg gcagtcaagc cgccattcac ccgggcagca
      361 tcacggggct ttgtaagcgc atggacgctg acgccgtgcc gacccggggc gagacgattg
      421 ggaagaagac cgcttcaagc gcctgggacc cggcaaccgt tatgcgaatc cttcgggacc
      481 cgcgtattgc gggcttcgcc gctgaggtga tctacaagaa gaagccggac ggcacgccga
      541 ccacgaagat tgagggttac cgcattcagc gcgacccgat cacgctccgg ccggtcgagc
      601 ttgattgcgg accgatcatc gagcccgctg agtggtatga gcttcaggcg tggttggacg
      661 gcagggggcg cggcaagggg ctttcccggg ggcaagccat tctgtccgcc atggacaagc
      721 tgtactgcga gtgtggcgcc gtcatgactt cgaagcgcgg ggaagaatcg atcaaggact
      781 cttaccgctg ccgtcgccgg aaggtggtcg acccgtccgc acctgggcag cacgaaggca
      841 cgtgcaacgt cagcatggcg gcactcgaca agttcgttgc ggaacgcatc ttcaacaaga
      901 tcaggcacgc cgaaggcgac gaagagacgt tggcgcttct gtgggaagcc gcccgacgct
      961 tcggcaagct cactgaggcg cctgagaaga gcggcgaacg ggcgaacctt gttgcggagc
     1021 gcgccgacgc cctgaacgcc cttgaagagc tgtacgaaga ccgcgcggca ggcgcgtacg
     1081 acggacccgt tggcaggaag cacttccgga agcaacaggc agcgctgacg ctccggcagc
     1141 aaggggcgga agagcggctt gccgaacttg aagccgccga agccccgaag cttccccttg
     1201 accaatggtt ccccgaagac gccgacgctg acccgaccgg ccctaagtcg tggtgggggc
     1261 gcgcgtcagt agacgacaag cgcgtgttcg tcgggctctt cgtagacaag atcgttgtca
     1321 cgaagtcgac tacgggcagg gggcagggaa cgcccatcga gaagcgcgct tcgatcacgt
     1381 gggcgaagcc gccgaccgac gacgacgaag acgacgccca ggacggcacg gaagacgtag
     1441 cggcgtagcg agacacccgg gaagcctgat ctacgtctgt cgagaagttt ctgatcgaaa
     1501 agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca
     1561 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct
     1621 acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc
     1681 ttgacattgg ggaatttatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     1741 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     1801 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     1861 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagcttgggc
     1921 tgcaggtcga ctctagtatg catgcgagtg tccgttcgag tggcggcttg cgcccgatgc
     1981 tagtcgcggt tgatcggcga tcgcaggtgc acgcggtcga tcttgacggc tggcgagagg
     2041 tgcggggagg atctgaccga cgcggtccac acgtggcacc gcgatgctgt tgtgggcaca
     2101 atcgtgccgg ttggtaggat ccacatatgg tgagcaaggg cgaggaggat aacatggcca
     2161 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
     2221 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     2281 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
     2341 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
     2401 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
     2461 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     2521 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
     2581 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     2641 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     2701 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     2761 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     2821 acgagctgta caagtagtct agaggatccg cggccgcgcg cgatatcgaa ttcgtaatca
     2881 tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     2941 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3001 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3061 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3121 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3181 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3241 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3301 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3361 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3421 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3481 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3541 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3601 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3661 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3721 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3781 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3841 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3901 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3961 ggatcttcac ctagatcctt ttggttcatg tgcagctcca tcagcaaaag gggatgataa
     4021 gtttatcacc accgactatt tgcaacagtg ccgttgatcg tgctatgatc gactgatgtc
     4081 atcagcggtg gagtgcaatg tcgtgcaata cgaatggcga aaagccgagc tcatcggtca
     4141 gcttctcaac cttggggtta cccccggcgg tgtgctgctg gtccacagct ccttccgtag
     4201 cgtccggccc ctcgaagatg ggccacttgg actgatcgag gccctgcgtg ctgcgctggg
     4261 tccgggaggg acgctcgtca tgccctcgtg gtcaggtctg gacgacgagc cgttcgatcc
     4321 tgccacgtcg cccgttacac cggaccttgg agttgtctct gacacattct ggcgcctgcc
     4381 aaatgtaaag cgcagcgccc atccatttgc ctttgcggca gcggggccac aggcagagca
     4441 gatcatctct gatccattgc ccctgccacc tcactcgcct gcaagcccgg tcgcccgtgt
     4501 ccatgaactc gatgggcagg tacttctcct cggcgtggga cacgatgcca acacgacgct
     4561 gcatcttgcc gagttgatgg caaaggttcc ctatggggtg ccgagacact gcaccattct
     4621 tcaggatggc aagttggtac gcgtcgatta tctcgagaat gaccactgct gtgagcgctt
     4681 tgccttggcg gacaggtggc tcaaggagaa gagccttcag aaggaaggtc cagtcggtca
     4741 tgcctttgct cggttgatcc gctcccgcga cattgtggcg acagccctgg gtcaactggg
     4801 ccgagatccg ttgatcttcc tgcatccgcc agaggcggga tgcgaagaat gcgatgccgc
     4861 tcgccagtcg attggctgag ctcatgagcg gagaacgaga tgacgttgga ggggcaaggt
     4921 cgcgctgatt gctggggcaa cacgtggagc ggatcgggga ttgtctttct tcagctcgct
     4981 gatgatatgc tgacgctcaa tgccgtttgg cctccgacta acgaaaatcc cgcatttgga
     5041 cggctgatcc gattggcacg gcggacggcg aatggcggag cagacgctcg tccgggggca
     5101 atgagatatg aaaaagcctg aactcaccgc gacgtatcgg gccctggcca gctagctaga
     5161 gtcgacctgc aggtccccgg ggatcggtct tgccttgctc gtcggtgatg tacttcacca
     5221 gctccgcgaa gtcgctcttc ttgatggagc gcatggggac gtgcttggca atcacgcgca
     5281 ccccccggcc gttttagcgg ctaaaaaagt catggctctg ccctcgggcg gaccacgccc
     5341 atcatgacct tgccaagctc gtcctgcttc tcttcgatct tcgccagcag ggcgaggatc
     5401 gtggcatcac cgaaccgcgc cgtgcgcggg tcgtcggtga gccagagttt cagcaggccg
     5461 cccaggcggc ccaggtcgcc attgatgcgg gccagctcgc ggacgtgctc atagtccacg
     5521 acgcccgtga ttttgtagcc ctggccgacg gccagcaggt aggccgacag gctcatgccg
     5581 gccgccgccg ccttttcctc aatcgctctt cgttcgtctg gaaggcagta caccttgata
     5641 ggtgggctgc ccttcctggt tggcttggtt tcatcagcca tccgcttgcc ctcatctgtt
     5701 acgccggcgg tagccggcca gcctcgcaga gcaggattcc cgttgagcac cgccaggtgc
     5761 gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg ggcctacttc acctatcctg
     5821 cccggctgac gccgttggat acaccaagga aagtctacac gaaccctttg gcaaaatcct
     5881 gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg ctataatgac cccgaagcag
     5941 ggttatgcag cggaaaagat ccgtcgacct gcaggcatgc aagctctagc gattccagac
     6001 gtcccgaagg cgtggcgcgg cttccccgtg ccggagcaat cgccctgggt gggttacacg
     6061 acgcccctct atggcccgta ctgacggaca caccgaagcc ccggcggcaa ccctcagcgg
     6121 atgccccggg gcttcacgtt ttcccaggtc agaagcggtt ttcgggagta gtgccccaac
     6181 tggggtaacc tttgagttct ctcagttggg ggcgtagggt cgccgacatg acacaagggg
     6241 ttgtgaccgg ggtggacacg tacgcgggtg cttacgaccg tcagtcgcgc gagcgcgaaa
     6301 attcgagcgc agcaagccca gcgacacagc gtagcgccaa cgaagacaag gcggccgacc
     6361 ttcagcgcga agtcgagcgc gacgggggcc ggttcaggtt cgtcgggcat ttcagcgaag
     6421 cgccgggcac gtcggcgttc gggacggcgg agcgcccgga gttcgaacgc atcctgaacg
     6481 aatgccgcgc cgggcggctc aacatgatca ttgtctatga cgtgtcgcgc ttctcgcgcc
     6541 tgaaggtcat ggacgcgatt ccgattgtct cggaattgct cgccctgggc gtgacgattg
     6601 tttccactca ggaaggcgtc t
//
