LOCUS       62056_10460        8927 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 8927)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..200
                     /label=AmpR promoter
     CDS             275..1462
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     rep_origin      1739..2327
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2615..2636
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2651..2681
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2689..2705
                     /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)."
     primer_bind     2713..2729
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3750..3778
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3967..4497
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     complement(5354..5370)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5720..5806
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      5898..5925
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     oriT            6369..6478
                     /label=oriT
                     /note="incP origin of transfer"
     promoter        6852..7297
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             7298..8716
                     /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"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aaccgattag ctttaatgcg gtagtttatc
      241 acagttaaat tgctaacgca gtcaggcacc gtgtatgaaa tctaacaatg cgctcatcgt
      301 catcctcggc accgtcaccc tggatgctgt aggcataggc ttggttatgc cggtactgcc
      361 gggcctcttg cgggatatcg tccattccga cagcatcgcc agtcactatg gcgtgctgct
      421 agcgctatat gcgttgatgc aatttctatg cgcacccgtt ctcggagcac tgtccgaccg
      481 ctttggccgc cgcccagtcc tgctcgcttc gctacttgga gccactatcg actacgcgat
      541 catggcgacc acacccgtcc tgtggattct ctacgccgga cgcatcgtgg ccggcatcac
      601 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
      661 ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt
      721 ggccggggga ctgttgggcg ccatctcctt acatgcacca ttccttgcgg cggcggtgct
      781 caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg
      841 ccgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
      901 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
      961 ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg
     1021 cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc
     1081 cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct
     1141 gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct
     1201 tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga
     1261 tgacgaccat cagggacagc ttcaaggatc gctcgcggct cttaccagcc taacttcgat
     1321 cactggaccg ctgatcgtca cggcgattta tgccgcctcg gcgagcacat ggaacgggtt
     1381 ggcatggatt gtaggcgccg ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc
     1441 atggagccgg gccacctcga cctgaatgga agccggcggc acctcgctaa cggattcacc
     1501 actccaagaa ttggagccaa tcaattcttg cggagaactg tgaatgcgca aaccaaccct
     1561 tgcatcgatt gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     1621 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     1681 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1741 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1801 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1861 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     1921 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     1981 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2041 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2101 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2161 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2221 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2281 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2341 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     2401 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     2461 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     2521 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     2581 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     2641 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     2701 aacaatttca cacaggaaac agctatgacc atgattacga attcagaccg tcaagccgcc
     2761 gacgctggag gagatcgagg cgatccgcca ggacgcctac aacgaaggct tcgccaccgg
     2821 cgagcgcgac ggtttccacg ccggccagtt gaaggcgcgg caggaagccg aggaggcgct
     2881 gaaggagcgt ctgcaaagtc tcgagcgtct gatgacgcag cttctggaac ccatcgccga
     2941 gcaggacgcc ctgatcgagc agggcatggt caacctggtc aaccacgttg cccgccaagt
     3001 gatccagcgc gaactgcaca tggactccag ccatgtccgc caggtcctgc gcgaggctct
     3061 gaagctgctg ccgatgggcg cggcgaacat ccgtatccat gtcaacccgc aggacttcga
     3121 gcgggtcaag gcgctgcgcg agcgccacga agagagctgg cggattctcg aggacgattc
     3181 gttgctgccc ggcggctgcc gcatcgagac cgagcacagt cgcatcgacg cgaccatcga
     3241 gacgcgcctg gcgcaggcag tgaagcaact gttcgagcaa cagcgcgaac aggcgaccca
     3301 tccgctggcc agcgacatcc ggatcgacct cgacgccccc ggagacgtcg atgcgccttg
     3361 agcgaaccag tttcgccagg cgcctggagg gctacaccga ggctgtctcg ctgccggcgc
     3421 agccggtggt cgagggccgc ctgctgcgca tggtcggcct gaccctcgaa gccgaaggct
     3481 tgcaggccgc ggtcggcagc cgctgcaacg tgatcaacga aagcggctac cacccggtgc
     3541 aggtggaggc cgaagtgatg ggcttttccg gtagcaaggt gtacctgatg ccggtcggca
     3601 gcctcgccgg catcgcgccc ggcgcgcggg tcgtgccgct gccggacacc gggcgcctgc
     3661 cgatgggcat gtcgatgctc ggccgcgtgc tcgacggcgc cggccgcgcc ctcgacggca
     3721 agggcgggat gcgcgccgaa gactgggtgt tgacataagc ctgttcggtt cgtaaactgt
     3781 aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg
     3841 gtaacggcgc agtggcggtt ttcatggctt gttatgactg tttttttgta cagtctatgc
     3901 ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca
     3961 gcaacgatgt tacgcagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag
     4021 ttaggtggct caagtatggg catcattcgc acatgtaggc tcggccctga ccaagtcaaa
     4081 tccatgcggg ctgctcttga tcttttcggt cgtgagttcg gagacgtagc cacctactcc
     4141 caacatcagc cggactccga ttacctcggg aacttgctcc gtagtaagac attcatcgcg
     4201 cttgctgcct tcgaccaaga agcggttgtt ggcgctctcg cggcttacgt tctgcccaag
     4261 tttgagcagc cgcgtagtga gatctatatc tatgatctcg cagtctccgg cgagcaccgg
     4321 aggcagggca ttgccaccgc gctcatcaat ctcctcaagc atgaggccaa cgcgcttggt
     4381 gcttatgtga tctacgtgca agcagattac ggtgacgatc ccgcagtggc tctctataca
     4441 aagttgggca tacgggaaga agtgatgcac tttgatatcg acccaagtac cgccacctaa
     4501 atcgaagctt cgatcagccg ggtgatgccg caggtggtcg aggccgagca cctgcgcgac
     4561 gcccagcgct tcaagcagct ctggtcgcgc taccagcaga gtcgcgacct gatcagcgtc
     4621 ggtgcctacg tggccggcgg cgatcccgag accgacctgg ccatcgcgcg cttcccggtc
     4681 atgcgccagt tcctgcgcca gggccttgac gaaagcgaaa gcctggcgga aagccgtgcg
     4741 cggctggcga gcctgctcgc cggcgggcag gcctgaggcg cgctgaggca tggagaagcg
     4801 cgcggcgcgg ctggcgccgg tggtggacat ggccagcaag gccgagcggg atgccgcgac
     4861 ccagctgggc aggtgccagc agcaattgct cgcggcacag cagaagctgg cggagctgga
     4921 gcgctaccgc aacgactacc agcagcaatg gatcagccag ggccagaagg gcgtcagcgg
     4981 ccagtggctg atgaactacc agcgcttcct ctcgcaactg gagaccgccg tcgcccagca
     5041 ggccaacagc gtgacctggc accgcgaggc ggtggacaag gcgcggctca actggcagga
     5101 gagatacgcc cgcctggaag ggctgcgcaa gctggtcgag cgctacctgg aagaggcgcg
     5161 ccaggccgag gacaagcgcg agcagaaaca gctcgacgaa ctcgcccagc gtacccgccg
     5221 ccaggacgac tgacgactca gaagaacgga cgcgacgcct tcagcatgaa tgtggtgtcg
     5281 cagtagctgt tgtgtcccga gtaggcgctg caatcgctgc cggtcaggct ggtgccgctg
     5341 taggaagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     5401 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     5461 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgagcttggc
     5521 tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc
     5581 ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
     5641 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga
     5701 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     5761 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
     5821 tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc
     5881 caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac
     5941 tctttttgtt tatttttcta aatacattca aatatgcgcc tgatgcggta ttttctcctt
     6001 acgcatctgt gcggtatttc acaccgcata tcgacatccg ccctcaccgc caggaacgca
     6061 accgcagcct catcacgccg gcgcttcttg gccgcgcggg attcaaccca ctcggccagc
     6121 tcgtcggtgt agctctttgg catcgtctct cgcctgtccc ctcagttcag taatttcctg
     6181 catttgcctg tttccagtcg gtagatattc cacaaaacag cagggaagca gcgcttttcc
     6241 gctgcataac cctgcttcgg ggtcattata gcgatttttt cggtatatcc atcctttttc
     6301 gcacgatata caggattttg ccaaagggtt cgtgtagact ttccttggtg tatccaacgg
     6361 cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc cttcttcact
     6421 gtcccttatt cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg ctggccggct
     6481 accgccggcg taacagatga gggcaagcgg atggctgatg aaaccaagcc aaccaggaag
     6541 ggcagcccac ctatcaaggt gtactgcctt ccagacgaac gaagagcgat tgaggaaaag
     6601 gcggcggcgg ccggcatgag cctgtcggcc tacctgctgg ccgtcggcca gggctacaaa
     6661 atcacgggcg tcgtggacta tgagcacgtc cgcgagctgg cccgcatcaa tggcgacctg
     6721 ggccgcctgg gcggcctgct gaaactctgg ctcaccgacg acccgcgcac ggcgcggttc
     6781 ggtgatgcca cgatcctcgc cctgctggcg aagatcgact ctagctagag gatcgatcct
     6841 ttttaaccca tcacatatac ctgccgttca ctattattta gtgaaatgag atattatgat
     6901 attttctgaa ttgtgattaa aaaggcaact ttatgcccat gcaacagaaa ctataaaaaa
     6961 tacagagaat gaaaagaaac agatagattt tttagttctt taggcccgta gtctgcaaat
     7021 ccttttatga ttttctatca aacaaaagag gaaaatagac cagttgcaat ccaaacgaga
     7081 gtctaataga atgaggtcga aaagtaaatc gcgcgggttt gttactgata aagcaggcaa
     7141 gacctaaaat gtgtaaaggg caaagtgtat actttggcgt caccccttac atattttagg
     7201 tcttttttta ttgtgcgtaa ctaacttgcc atcttcaaac aggagggctg gaagaagcag
     7261 accgctaaca cagtacataa aaaaggagac atgaacgatg aacatcaaaa agtttgcaaa
     7321 acaagcaaca gtattaacct ttactaccgc actgctggca ggaggcgcaa ctcaagcgtt
     7381 tgcgaaagaa acgaaccaaa agccatataa ggaaacatac ggcatttccc atattacacg
     7441 ccatgatatg ctgcaaatcc ctgaacagca aaaaaatgaa aaatatcaag ttcctgagtt
     7501 cgattcgtcc acaattaaaa atatctcttc tgcaaaaggc ctggacgttt gggacagctg
     7561 gccattacaa aacgctgacg gcactgtcgc aaactatcac ggctaccaca tcgtctttgc
     7621 attagccgga gatcctaaaa atgcggatga cacatcgatt tacatgttct atcaaaaagt
     7681 cggcgaaact tctattgaca gctggaaaaa cgctggccgc gtctttaaag acagcgacaa
     7741 attcgatgca aatgattcta tcctaaaaga ccaaacacaa gaatggtcag gttcagccac
     7801 atttacatct gacggaaaaa tccgtttatt ctacactgat ttctccggta aacattacgg
     7861 caaacaaaca ctgacaactg cacaagttaa cgtatcagca tcagacagct ctttgaacat
     7921 caacggtgta gaggattata aatcaatctt tgacggtgac ggaaaaacgt atcaaaatgt
     7981 acagcagttc atcgatgaag gcaactacag ctcaggcgac aaccatacgc tgagagatcc
     8041 tcactacgta gaagataaag gccacaaata cttagtattt gaagcaaaca ctggaactga
     8101 agatggctac caaggcgaag aatctttatt taacaaagca tactatggca aaagcacatc
     8161 attcttccgt caagaaagtc aaaaacttct gcaaagcgat aaaaaacgca cggctgagtt
     8221 agcaaacggc gctctcggta tgattgagct aaacgatgat tacacactga aaaaagtgat
     8281 gaaaccgctg attgcatcta acacagtaac agatgaaatt gaacgcgcga acgtctttaa
     8341 aatgaacggc aaatggtatc tgttcactga ctcccgcgga tcaaaaatga cgattgacgg
     8401 cattacgtct aacgatattt acatgcttgg ttatgtttct aattctttaa ctggcccata
     8461 caagccgctg aacaaaactg gccttgtgtt aaaaatggat cttgatccta acgatgtaac
     8521 ctttacttac tcacacttcg ctgtacctca agcgaaagga aacaatgtcg tgattacaag
     8581 ctatatgaca aacagaggat tctacgcaga caaacaatca acgtttgcgc ctagcttcct
     8641 gctgaacatc aaaggcaaga aaacatctgt tgtcaaagac agcatccttg aacaaggaca
     8701 attaacagtt aacaaataaa aacgcaaaag aaaatgccga ttatggtgca ctctcagtac
     8761 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     8821 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     8881 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcga
//
