LOCUS       62056_12695        6633 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 6633)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6633
                     /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             2130..2846
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2894..2910)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2918..2934)
                     /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(2942..2972)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2987..3008)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3296..3884)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             4088..4888
                     /codon_start=1
                     /label=ApmR
                     /note="aminoglycoside 3-N-acetyltransferase type IV"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     CDS             complement(5325..5693)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(5726..5835)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(6073..6084)
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     protein_bind    6155..6254
                     /label=phage phi-C31 attP
                     /note="attachment site of phage phi-C31"
     CDS             join(6264..6633,1..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 ccacatatga tggtctccaa gggcgaggag ctgttcaccg
     2161 gcgtcgtccc gatcctggtc gagctggacg gcgacgtcaa cggccacaag ttctccgtct
     2221 ccggcgaggg cgagggcgac gccacctacg gcaagctgac cctgaagttc atctgcacca
     2281 ccggcaagct gccggtcccg tggccgaccc tggtcaccac cctgacctac ggcgtccagt
     2341 gcttctcccg ctacccggac cacatgaagc agcacgactt cttcaagtcc gccatgccgg
     2401 agggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     2461 ccgaggtcaa gttcgagggc gacaccctgg tcaaccgcat cgagctgaag ggcatcgact
     2521 tcaaggagga cggcaacatc ctgggccaca agctggagta caactacaac tcccacaacg
     2581 tctacatcat ggccgacaag cagaagaacg gcatcaaggt caacttcaag atccgccaca
     2641 acatcgagga cggctccgtc cagctggccg accactacca gcagaacacc ccgatcggcg
     2701 acggcccggt cctgctgccg gacaaccact acctgtccac ccagtccgcc ctgtccaagg
     2761 acccgaacga gaagcgcgac cacatggtcc tgctggagtt cgtcaccgcc gccggcatca
     2821 ccctgggcat ggacgagctg tacaagtaat ctagaggatc cgcggccgcg cgcgatatcg
     2881 aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2941 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3001 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3061 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3121 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3181 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3241 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3301 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3361 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3421 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3481 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3541 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3601 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3661 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3721 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3781 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3841 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3901 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3961 gattatcaaa aaggatcttc acctagatcc ttttggttca tgtgcagctc catcagcaaa
     4021 aggggatgat aagtttatca ccaccgacta tttgcaacag tgccgttgat cgtgctatga
     4081 tcgactgatg tcatcagcgg tggagtgcaa tgtcgtgcaa tacgaatggc gaaaagccga
     4141 gctcatcggt cagcttctca accttggggt tacccccggc ggtgtgctgc tggtccacag
     4201 ctccttccgt agcgtccggc ccctcgaaga tgggccactt ggactgatcg aggccctgcg
     4261 tgctgcgctg ggtccgggag ggacgctcgt catgccctcg tggtcaggtc tggacgacga
     4321 gccgttcgat cctgccacgt cgcccgttac accggacctt ggagttgtct ctgacacatt
     4381 ctggcgcctg ccaaatgtaa agcgcagcgc ccatccattt gcctttgcgg cagcggggcc
     4441 acaggcagag cagatcatct ctgatccatt gcccctgcca cctcactcgc ctgcaagccc
     4501 ggtcgcccgt gtccatgaac tcgatgggca ggtacttctc ctcggcgtgg gacacgatgc
     4561 caacacgacg ctgcatcttg ccgagttgat ggcaaaggtt ccctatgggg tgccgagaca
     4621 ctgcaccatt cttcaggatg gcaagttggt acgcgtcgat tatctcgaga atgaccactg
     4681 ctgtgagcgc tttgccttgg cggacaggtg gctcaaggag aagagccttc agaaggaagg
     4741 tccagtcggt catgcctttg ctcggttgat ccgctcccgc gacattgtgg cgacagccct
     4801 gggtcaactg ggccgagatc cgttgatctt cctgcatccg ccagaggcgg gatgcgaaga
     4861 atgcgatgcc gctcgccagt cgattggctg agctcatgag cggagaacga gatgacgttg
     4921 gaggggcaag gtcgcgctga ttgctggggc aacacgtgga gcggatcggg gattgtcttt
     4981 cttcagctcg ctgatgatat gctgacgctc aatgccgttt ggcctccgac taacgaaaat
     5041 cccgcatttg gacggctgat ccgattggca cggcggacgg cgaatggcgg agcagacgct
     5101 cgtccggggg caatgagata tgaaaaagcc tgaactcacc gcgacgtatc gggccctggc
     5161 cagctagcta gagtcgacct gcaggtcccc ggggatcggt cttgccttgc tcgtcggtga
     5221 tgtacttcac cagctccgcg aagtcgctct tcttgatgga gcgcatgggg acgtgcttgg
     5281 caatcacgcg caccccccgg ccgttttagc ggctaaaaaa gtcatggctc tgccctcggg
     5341 cggaccacgc ccatcatgac cttgccaagc tcgtcctgct tctcttcgat cttcgccagc
     5401 agggcgagga tcgtggcatc accgaaccgc gccgtgcgcg ggtcgtcggt gagccagagt
     5461 ttcagcaggc cgcccaggcg gcccaggtcg ccattgatgc gggccagctc gcggacgtgc
     5521 tcatagtcca cgacgcccgt gattttgtag ccctggccga cggccagcag gtaggccgac
     5581 aggctcatgc cggccgccgc cgccttttcc tcaatcgctc ttcgttcgtc tggaaggcag
     5641 tacaccttga taggtgggct gcccttcctg gttggcttgg tttcatcagc catccgcttg
     5701 ccctcatctg ttacgccggc ggtagccggc cagcctcgca gagcaggatt cccgttgagc
     5761 accgccaggt gcgaataagg gacagtgaag aaggaacacc cgctcgcggg tgggcctact
     5821 tcacctatcc tgcccggctg acgccgttgg atacaccaag gaaagtctac acgaaccctt
     5881 tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat cgctataatg
     5941 accccgaagc agggttatgc agcggaaaag atccgtcgac ctgcaggcat gcaagctcta
     6001 gcgattccag acgtcccgaa ggcgtggcgc ggcttccccg tgccggagca atcgccctgg
     6061 gtgggttaca cgacgcccct ctatggcccg tactgacgga cacaccgaag ccccggcggc
     6121 aaccctcagc ggatgccccg gggcttcacg ttttcccagg tcagaagcgg ttttcgggag
     6181 tagtgcccca actggggtaa cctttgagtt ctctcagttg ggggcgtagg gtcgccgaca
     6241 tgacacaagg ggttgtgacc ggggtggaca cgtacgcggg tgcttacgac cgtcagtcgc
     6301 gcgagcgcga aaattcgagc gcagcaagcc cagcgacaca gcgtagcgcc aacgaagaca
     6361 aggcggccga ccttcagcgc gaagtcgagc gcgacggggg ccggttcagg ttcgtcgggc
     6421 atttcagcga agcgccgggc acgtcggcgt tcgggacggc ggagcgcccg gagttcgaac
     6481 gcatcctgaa cgaatgccgc gccgggcggc tcaacatgat cattgtctat gacgtgtcgc
     6541 gcttctcgcg cctgaaggtc atggacgcga ttccgattgt ctcggaattg ctcgccctgg
     6601 gcgtgacgat tgtttccact caggaaggcg tct
//
