LOCUS       62056_9185       10295 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 10295)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             38..694
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQSHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     oriT            3421..3530
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             3563..3931
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     promoter        complement(4540..4558)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(4627..4657)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4672..4693)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4981..5569)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5743..6600)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6601..6705)
                     /label=AmpR promoter
     rep_origin      complement(6731..7186)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        7354..7372
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        7545..7990
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             7991..9409
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     CDS             complement(10072..10083)
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
ORIGIN
        1 agcttcgacg agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata
       61 taccaccgtt gatatatccc aatcgcatcg taaagaacat tttgaggcat ttcagtcagt
      121 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
      181 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
      241 tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
      301 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
      361 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
      421 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
      481 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
      541 ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
      601 gattcaggtt catcatgccg tctgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
      661 acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg
      721 cccttaaacg cctggtgcta cgcctgaata agtgataata agcggatgaa tggcagaaat
      781 tcgtcgaagc ttgatgcagg aattcgatat caagcttatc gataccgtcg aaattcccgg
      841 gagcgggccc ccccctcgag gtcgacggta tcgataagct tgcatgcctg cattttaaac
      901 ttaatcccta tgcgttagcg tttacttcgc tgtttcttgt cgcttgttct ggcggaaaag
      961 gaagttttga tttagaagat gtccggccta ataagacaac aggcgtgtct aaagaggagt
     1021 acaaggatgt agaaacagcc aagaaagaaa aagaacagtt aggggaatta atggaacctg
     1081 ctttggggta tgttgtaaaa gttccggtga gttcttttga aaataagaaa gttgatattt
     1141 cagatataga agtgattacg aacggaaatt tagacgatgt gccgtacaag gcaaattcat
     1201 ctaaatataa ctatccagat ataaaaacaa aagattcttc tcttcagtac gttcgctcag
     1261 gatatgttat tgatggggaa cactctggtt ctaatgaaaa gggatatgtg tattataaag
     1321 gtaattcacc tgcaaaagaa ttacccgtta atcagctttt aacttataca ggaagttggg
     1381 attttacttc caatgcgaat ttaaataatg aagagggaag acctaattat ttaaacgacg
     1441 attattatac taaatttata ggtaaacggg tgggcttggt ttcgggagat gcgaaacctg
     1501 caaagcataa atacactagc cagtttgaag ttgattttgc aactaaaaaa atgacaggta
     1561 aattatccga taaagagaaa acgatttata cagtcaatgc tgatattaga ggcaatcgtt
     1621 ttacgggggc tgctacagcg agtgataaaa ataaagggaa aggcgaatca tataacttct
     1681 ttagtgccga ttctcagtct ttagaaggcg gcttctatgg tccaaaagca gaagaaatgg
     1741 cagggaaatt tgtagctaac gacaaatctc tttttgccgt tttttcagca aaacacaatg
     1801 gctctaatgt taacaccgtt cggattattg atgcctcaaa aattgattta actaatttca
     1861 gcatttcaga acttaacaat tttggtgatg cttccgtttt aattattgat gggaaaaaaa
     1921 taaagctagc tggtagcggg tttacaaata agcacactat tgaaatcaat ggcaaaacaa
     1981 tggtagccgt agcctgctgt agtaatctgg aatatatgaa gtttggtcaa ttatggcaac
     2041 aagcagaggg cggaaaaccc gagaataata gtttattcct acaaggcgaa cgtaccgcaa
     2101 cagataagat gccaaaaggc ggaaactata aatatattgg tacttgggat gctcaggttt
     2161 caaaagaaaa taactgggtt gctacggcag atgatgatag aaaagctggc tatcggacag
     2221 aatttgatgt tgattttggc aacaaaaatt taagtggtaa gttatttgat aaaaacggtg
     2281 taaatcctgt gtttaccgta gatgcaaaaa ttgatggtaa tggttttact ggcaaagcta
     2341 aaacctcaga tgaaggcttc gctctagatt caggtagttc acgttatgag aatgtgaaat
     2401 ttaacgatgt agcagttagt ggtggcttct atggtccaac ggcagcagag cttggcggac
     2461 aattccacca taaatcagaa aatggcagtg taggtgctgt ctttggtgca aaacaacaag
     2521 taaaaaaata ataaggaatt tgcaatgaaa aataaattaa atctgattag ccttgctctg
     2581 cttagcctct ttgccgtaca aagctatgca gaacaagcgg tgcaattgaa cgatgtttat
     2641 gtcacaggta ccaaaaagaa agcacataaa aaagagaacg aagtgacagg cttagggaaa
     2701 gtagtgaaaa caccagattc tcttagtaag gagcaagtgt taggaatgcg agatccccgg
     2761 gggatccact agttctagag cggccgccac cgcggtggga tcctttttgt ccggtgttgg
     2821 gttgaaggtg aagccggtcg gggccgcagc gggggccggc ttttcagcct tgcccccctg
     2881 cttcggccgc cgtggctccg gcgtcttggg tgccggcgcg ggttccgcag ccttggcctg
     2941 cggtgcgggc acatcggcgg gcttggcctt gatgtgccgc ctggcgtgcg agcggaacgt
     3001 ctcgtaggag aacttgacct tccccgtttc ccgcatgtgc tcccaaatgg tgacgagcgc
     3061 atagccggac gctaacgccg cctcgacatc cgccctcacc gccaggaacg caaccgcagc
     3121 ctcatcacgc cggcgcttct tggccgcgcg ggattcaacc cactcggcca gctcgtcggt
     3181 gtagctcttt ggcatcgtct ctcgcctgtc ccctcagttc agtaatttcc tgcatttgcc
     3241 tgtttccagt cggtagatat tccacaaaac agcagggaag cagcgctttt ccgctgcata
     3301 accctgcttc ggggtcatta tagcgatttt ttcggtatat ccatcctttt tcgcacgata
     3361 tacaggattt tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac ggcgtcagcc
     3421 gggcaggata ggtgaagtag gcccacccgc gagcgggtgt tccttcttca ctgtccctta
     3481 ttcgcacctg gcggtgctca acgggaatcc tgctctgcga ggctggccgg ctaccgccgg
     3541 cgtaacagat gagggcaagc ggatggctga tgaaaccaag ccaaccagga agggcagccc
     3601 acctatcaag gtgtactgcc ttccagacga acgaagagcg attgaggaaa aggcggcggc
     3661 ggccggcatg agcctgtcgg cctacctgct ggccgtcggc cagggctaca aaatcacggg
     3721 cgtcgtggac tatgagcacg tccgcgagct ggcccgcatc aatggcgacc tgggccgcct
     3781 gggcggcctg ctgaaactct ggctcaccga cgacccgcgc acggcgcggt tcggtgatgc
     3841 cacgatcctc gccctgctgg cgaagatcga agagaagcag gacgagcttg gcaaggtcat
     3901 gatgggcgtg gtccgcccga gggcagagcc atgacttttt tagccgctaa aacggccggg
     3961 gggtgcgcgt gattgccaag cacgtcccca tgcgctccat caagaagagc gacttcgcgg
     4021 agctggtgaa gtacatcacc gacgagcaag gcaagaccga gcgcctgggt cacgtgcgcg
     4081 tcacgaactg cgaggcaaac accctgcccg ctgtcatggc cgaggtgatg gcgacccagc
     4141 acggcaacac ccgttccgag gccgacaaga cctatcacct gctggttagc ttccgcgcgg
     4201 gagagaagcc cgacgcggag acgttgcgcg cgattgagga ccgcatctgc gctgggcttg
     4261 gcttcgccga gcatcagcgc gtcagtgccg tgcatcacga caccgacaac ctgcacatcc
     4321 atatcgccat caacaagatt cacccgaccc gaaacaccat ccatgagccg tatcgggcct
     4381 accgcgccct cgctgacctc tgcgcgacgc tcgaacggga ctacgggctt gagcgtgaca
     4441 atcacgaaac gcggcagcgc gtttccgaga accgcgcgaa cgacatggag cggcacgcgg
     4501 gcgtggaaag cctggtcggc tggatcccag cttttgttcc ctttagtgag ggttaattgc
     4561 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4621 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4681 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4741 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4801 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4861 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4921 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4981 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5041 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5101 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5161 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5221 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5281 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5341 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5401 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     5461 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5521 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5581 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5641 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5701 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5761 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5821 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5881 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5941 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6001 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6061 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6121 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6181 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6241 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6301 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6361 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6421 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6481 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6541 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6601 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6661 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6721 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     6781 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     6841 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     6901 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     6961 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     7021 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     7081 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     7141 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
     7201 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     7261 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     7321 cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg
     7381 ggatccgctt ttacagcgat tgcagaatga ttgaattgta aactttagag ctttatattt
     7441 tgtttaatgg tattatattt acttatattt atgattctta gtttttattg taaattaaag
     7501 tgtttattta ttgtatttta agtataagat cctttttaac ccatcacata tacctgccgt
     7561 tcactattat ttagtgaaat gagatattat gatattttct gaattgtgat taaaaaggca
     7621 actttatgcc catgcaacag aaactataaa aaatacagag aatgaaaaga aacagataga
     7681 ttttttagtt ctttaggccc gtagtctgca aatcctttta tgattttcta tcaaacaaaa
     7741 gaggaaaata gaccagttgc aatccaaacg agagtctaat agaatgaggt cgaaaagtaa
     7801 atcgcgcggg tttgttactg ataaagcagg caagacctaa aatgtgtaaa gggcaaagtg
     7861 tatactttgg cgtcacccct tacatatttt aggtcttttt ttattgtgcg taactaactt
     7921 gccatcttca aacaggaggg ctggaagaag cagaccgcta acacagtaca taaaaaagga
     7981 gacatgaacg atgaacatca aaaagtttgc aaaacaagca acagtattaa cctttactac
     8041 cgcactgctg gcaggaggcg caactcaagc gtttgcgaaa gaaacgaacc aaaagccata
     8101 taaggaaaca tacggcattt cccatattac acgccatgat atgctgcaaa tccctgaaca
     8161 gcaaaaaaat gaaaaatatc aagttcctga attcgattcg tccacaatta aaaatatctc
     8221 ttctgcaaaa ggcctggacg tttgggacag ctggccatta caaaacgctg acggcactgt
     8281 cgcaaactat cacggctacc acatcgtctt tgcattagcc ggagatccta aaaatgcgga
     8341 tgacacatcg atttacatgt tctatcaaaa agtcggcgaa acttctattg acagctggaa
     8401 aaacgctggc cgcgtcttta aagacagcga caaattcgat gcaaatgatt ctatcctaaa
     8461 agaccaaaca caagaatggt caggttcagc cacatttaca tctgacggaa aaatccgttt
     8521 attctacact gatttctccg gtaaacatta cggcaaacaa acactgacaa ctgcacaagt
     8581 taacgtatca gcatcagaca gctctttgaa catcaacggt gtagaggatt ataaatcaat
     8641 ctttgacggt gacggaaaaa cgtatcaaaa tgtacagcag ttcatcgatg aaggcaacta
     8701 cagctcaggc gacaaccata cgctgagaga tcctcactac gtagaagata aaggccacaa
     8761 atacttagta tttgaagcaa acactggaac tgaagatggc taccaaggcg aagaatcttt
     8821 atttaacaaa gcatactatg gcaaaagcac atcattcttc cgtcaagaaa gtcaaaaact
     8881 tctgcaaagc gataaaaaac gcacggctga gttagcaaac ggcgctctcg gtatgattga
     8941 gctaaacgat gattacacac tgaaaaaagt gatgaaaccg ctgattgcat ctaacacagt
     9001 aacagatgaa attgaacgcg cgaacgtctt taaaatgaac ggcaaatggt acctgttcac
     9061 tgactcccgc ggatcaaaaa tgacgattga cggcattacg tctaacgata tttacatgct
     9121 tggttatgtt tctaattctt taactggccc atacaagccg ctgaacaaaa ctggccttgt
     9181 gttaaaaatg gatcttgatc ctaacgatgt aacctttact tactcacact tcgctgtacc
     9241 tcaagcgaaa ggaaacaatg tcgtgattac aagctatatg acaaacagag gattctacgc
     9301 agacaaacaa tcaacgtttg cgccaagctt cctgctgaac atcaaaggca agaaaacatc
     9361 tgttgtcaaa gacagcatcc ttgaacaagg acaattaaca gttaacaaat aaaaacgcaa
     9421 aagaaaatgc cgatatccta ttggcatttt cttttatttc ttatcaacat aaaggtgaat
     9481 cccatacctc gagcttcacg ctgccgcaag cactcagggc gcaagggctg ctaaaggaag
     9541 cggaacacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac
     9601 tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg
     9661 cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca
     9721 gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg
     9781 ccgccaagga tctgatggcg caggggatca agatctgatc aagagacagg atgaggatcg
     9841 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     9901 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     9961 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
    10021 gaactgcatg ctcgagcggc cggccgccag tgtgatggat atctgcagaa ttcgcccttc
    10081 gatggatccg cttttacagc gattgcagaa tgattgaatt gtaaacttta gagctttata
    10141 ttttgtttaa tggtattata tttacttata tttatgattc ttagttttta ttgtaaatta
    10201 aagtgtttat ttattgtatt ttaagtataa gatctcggca agggcgaatt ccagcacact
    10261 ggcggccgtt actagtggat ccgagctcgg tacca
//
