LOCUS       62056_10455        7060 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 7060)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7060
                     /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"
     CDS             2744..3460
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3510..3526)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3876..3962
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4054..4081
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     oriT            4502..4611
                     /label=oriT
                     /note="incP origin of transfer"
     promoter        4985..5430
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             5431..6849
                     /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 agctatgaca tgattacgaa ttcatggtga gcaagggcga
     2761 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2821 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2881 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2941 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     3001 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     3061 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     3121 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     3181 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     3241 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     3301 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     3361 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     3421 cgccgccggg atcactctcg gcatggacga gctgtacaag aggtacccgg ggatcctcta
     3481 gagtcgacct gcaggcatgc aagcttggca ctggccgtcg ttttacaacg tcgtgactgg
     3541 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     3601 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     3661 gaatggcgag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
     3721 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
     3781 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
     3841 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     3901 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     3961 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     4021 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     4081 ttgcgtttct acaaactctt tttgtttatt tttctaaata cattcaaata tgcgcctgat
     4141 gcggtatttt ctccttacgc atatcgacat ccgccctcac cgccaggaac gcaaccgcag
     4201 cctcatcacg ccggcgcttc ttggccgcgc gggattcaac ccactcggcc agctcgtcgg
     4261 tgtagctctt tggcatcgtc tctcgcctgt cccctcagtt cagtaatttc ctgcatttgc
     4321 ctgtttccag tcggtagata ttccacaaaa cagcagggaa gcagcgcttt tccgctgcat
     4381 aaccctgctt cggggtcatt atagcgattt tttcggtata tccatccttt ttcgcacgat
     4441 atacaggatt ttgccaaagg gttcgtgtag actttccttg gtgtatccaa cggcgtcagc
     4501 cgggcaggat aggtgaagta ggcccacccg cgagcgggtg ttccttcttc actgtccctt
     4561 attcgcacct ggcggtgctc aacgggaatc ctgctctgcg aggctggccg gctaccgccg
     4621 gcgtaacaga tgagggcaag cggatggctg atgaaaccaa gccaaccagg aagggcagcc
     4681 cacctatcaa ggtgtactgc cttccagacg aacgaagagc gattgaggaa aaggcggcgg
     4741 cggccggcat gagcctgtcg gcctacctgc tggccgtcgg ccagggctac aaaatcacgg
     4801 gcgtcgtgga ctatgagcac gtccgcgagc tggcccgcat caatggcgac ctgggccgcc
     4861 tgggcggcct gctgaaactc tggctcaccg acgacccgcg cacggcgcgg ttcggtgatg
     4921 ccacgatcct cgccctgctg gcgaagatcg actctagcta gaggatcgat cctttttaac
     4981 ccatcacata tacctgccgt tcactattat ttagtgaaat gagatattat gatattttct
     5041 gaattgtgat taaaaaggca actttatgcc catgcaacag aaactataaa aaatacagag
     5101 aatgaaaaga aacagataga ttttttagtt ctttaggccc gtagtctgca aatcctttta
     5161 tgattttcta tcaaacaaaa gaggaaaata gaccagttgc aatccaaacg agagtctaat
     5221 agaatgaggt cgaaaagtaa atcgcgcggg tttgttactg ataaagcagg caagacctaa
     5281 aatgtgtaaa gggcaaagtg tatactttgg cgtcacccct tacatatttt aggtcttttt
     5341 ttattgtgcg taactaactt gccatcttca aacaggaggg ctggaagaag cagaccgcta
     5401 acacagtaca taaaaaagga gacatgaacg atgaacatca aaaagtttgc aaaacaagca
     5461 acagtattaa cctttactac cgcactgctg gcaggaggcg caactcaagc gtttgcgaaa
     5521 gaaacgaacc aaaagccata taaggaaaca tacggcattt cccatattac acgccatgat
     5581 atgctgcaaa tccctgaaca gcaaaaaaat gaaaaatatc aagttcctga gttcgattcg
     5641 tccacaatta aaaatatctc ttctgcaaaa ggcctggacg tttgggacag ctggccatta
     5701 caaaacgctg acggcactgt cgcaaactat cacggctacc acatcgtctt tgcattagcc
     5761 ggagatccta aaaatgcgga tgacacatcg atttacatgt tctatcaaaa agtcggcgaa
     5821 acttctattg acagctggaa aaacgctggc cgcgtcttta aagacagcga caaattcgat
     5881 gcaaatgatt ctatcctaaa agaccaaaca caagaatggt caggttcagc cacatttaca
     5941 tctgacggaa aaatccgttt attctacact gatttctccg gtaaacatta cggcaaacaa
     6001 acactgacaa ctgcacaagt taacgtatca gcatcagaca gctctttgaa catcaacggt
     6061 gtagaggatt ataaatcaat ctttgacggt gacggaaaaa cgtatcaaaa tgtacagcag
     6121 ttcatcgatg aaggcaacta cagctcaggc gacaaccata cgctgagaga tcctcactac
     6181 gtagaagata aaggccacaa atacttagta tttgaagcaa acactggaac tgaagatggc
     6241 taccaaggcg aagaatcttt atttaacaaa gcatactatg gcaaaagcac atcattcttc
     6301 cgtcaagaaa gtcaaaaact tctgcaaagc gataaaaaac gcacggctga gttagcaaac
     6361 ggcgctctcg gtatgattga gctaaacgat gattacacac tgaaaaaagt gatgaaaccg
     6421 ctgattgcat ctaacacagt aacagatgaa attgaacgcg cgaacgtctt taaaatgaac
     6481 ggcaaatggt atctgttcac tgactcccgc ggatcaaaaa tgacgattga cggcattacg
     6541 tctaacgata tttacatgct tggttatgtt tctaattctt taactggccc atacaagccg
     6601 ctgaacaaaa ctggccttgt gttaaaaatg gatcttgatc ctaacgatgt aacctttact
     6661 tactcacact tcgctgtacc tcaagcgaaa ggaaacaatg tcgtgattac aagctatatg
     6721 acaaacagag gattctacgc agacaaacaa tcaacgtttg cgcctagctt cctgctgaac
     6781 atcaaaggca agaaaacatc tgttgtcaaa gacagcatcc ttgaacaagg acaattaaca
     6841 gttaacaaat aaaaacgcaa aagaaaatgc cgattatggt gcactctcag tacaatctgc
     6901 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     6961 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     7021 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
//
