LOCUS       62056_13220        5381 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 5381)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5381
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             74..1492
                     /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(1591..2121)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(2310..2338)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     primer_bind     2877..2893
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2903..2921
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(2930..3037)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(3050..3068)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3089..3105)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3113..3129)
                     /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(3137..3167)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3182..3203)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3596..4141)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     oriT            complement(4524..4633)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     promoter        5009..5381
                     /label=sacB promoter
                     /note="sacB promoter and control region"
ORIGIN
        1 cttgccatct tcaaacagga gggctggaag aagcagaccg ctaacacagt acataaaaaa
       61 ggagacatga acgatgaaca tcaaaaagtt tgcaaaacaa gcaacagtat taacctttac
      121 taccgcactg ctggcaggag gcgcaactca agcgtttgcg aaagaaacga accaaaagcc
      181 atataaggaa acatacggca tttcccatat tacacgccat gatatgctgc aaatccctga
      241 acagcaaaaa aatgaaaaat atcaagttcc tgaattcgat tcgtccacaa ttaaaaatat
      301 ctcttctgca aaaggcctgg acgtttggga cagctggcca ttacaaaacg ctgacggcac
      361 tgtcgcaaac tatcacggct accacatcgt ctttgcatta gccggagatc ctaaaaatgc
      421 ggatgacaca tcgatttaca tgttctatca aaaagtcggc gaaacttcta ttgacagctg
      481 gaaaaacgct ggccgcgtct ttaaagacag cgacaaattc gatgcaaatg attctatcct
      541 aaaagaccaa acacaagaat ggtcaggttc agccacattt acatctgacg gaaaaatccg
      601 tttattctac actgatttct ccggtaaaca ttacggcaaa caaacactga caactgcaca
      661 agttaacgta tcagcatcag acagctcttt gaacatcaac ggtgtagagg attataaatc
      721 aatctttgac ggtgacggaa aaacgtatca aaatgtacag cagttcatcg atgaaggcaa
      781 ctacagctca ggcgacaacc atacgctgag agatcctcac tacgtagaag ataaaggcca
      841 caaatactta gtatttgaag caaacactgg aactgaagat ggctaccaag gcgaagaatc
      901 tttatttaac aaagcatact atggcaaaag cacatcattc ttccgtcaag aaagtcaaaa
      961 acttctgcaa agcgataaaa aacgcacggc tgagttagca aacggcgctc tcggtatgat
     1021 tgagctaaac gatgattaca cactgaaaaa agtgatgaaa ccgctgattg catctaacac
     1081 agtaacagat gaaattgaac gcgcgaacgt ctttaaaatg aacggcaaat ggtacctgtt
     1141 cactgactcc cgcggatcaa aaatgacgat tgacggcatt acgtctaacg atatttacat
     1201 gcttggttat gtttctaatt ctttaactgg cccatacaag ccgctgaaca aaactggcct
     1261 tgtgttaaaa atggatcttg atcctaacga tgtaaccttt acttactcac acttcgctgt
     1321 acctcaagcg aaaggaaaca atgtcgtgat tacaagctat atgacaaaca gaggattcta
     1381 cgcagacaaa caatcaacgt ttgcgccaag cttcctgctg aacatcaaag gcaagaaaac
     1441 atctgttgtc aaagacagca tccttgaaca aggacaatta acagttaaca aataaaaacg
     1501 caaaagaaaa tgccgatggc cgcggcgttg tgacaattta ccgaacaact ccgcggccgg
     1561 gaagccgatc tcggcttgaa cgaattgtta ggtggcggta cttgggtcga tatcaaagtg
     1621 catcacttct tcccgtatgc ccaactttgt atagagagcc actgcgggat cgtcaccgta
     1681 atctgcttgc acgtagatca cataagcacc aagcgcgttg gcctcatgct tgaggagatt
     1741 gatgagcgcg gtggcaatgc cctgcctccg gtgctcgccg gagactgcga gatcatagat
     1801 atagatctca ctacgcggct gctcaaactt gggcagaacg taagccgcga gagcgccaac
     1861 aaccgcttct tggtcgaagg cagcaagcgc gatgaatgtc ttactacgga gcaagttccc
     1921 gaggtaatcg gagtccggct gatgttggga gtaggtggct acgtctccga actcacgacc
     1981 gaaaagatca agagcagccc gcatggattt gacttggtca gggccgagcc tacatgtgcg
     2041 aatgatgccc atacttgagc cacctaactt tgttttaggg cgactgccct gctgcgtaac
     2101 atcgttgctg ctgcgtaaca tcgttgctgc tccataacat caaacatcga cccacggcgt
     2161 aacgcgcttg ctgcttggat gcccgaggca tagactgtac aaaaaaacag tcataacaag
     2221 ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt
     2281 tgcgtgagcg catacgctac ttgcattaca gtttacgaac cgaacaggct tatgtcaact
     2341 gggttcgtgc cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg ggcagcagcg
     2401 aagtcgaggc atttctgtcc tggctggcga acgagcgcaa ggtttcggtc tccacgcatc
     2461 gtcaggcatt ggcggccttg ctgttcttct acggcaaggt gctgtgcacg gatctgccct
     2521 ggcttcagga gatcggaaga cctcggccgt cgcggcgctt gccggtggtg ctgaccccgg
     2581 atgaagtggt tcgcatcctc ggttttctgg aaggcgagca tcgtttgttc gcccagcttc
     2641 tgtatggaac gggcatgcgg atcagtgagg gtttgcaact gcgggtcaag gatctggatt
     2701 tcgatcacgg cacgatcatc gtgcgggagg gcaagggctc caaggatcgg gccttgatgt
     2761 tacccgagag cttggcaccc agcctgcgcg agcagaatta attcccctgg cgaaaggggg
     2821 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     2881 aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg gtaccgggcc
     2941 ccccctcgag gtcgacggta tcgataagct tgatatcgaa ttcctgcagc ccgggggatc
     3001 cactagttct agagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag
     3061 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     3121 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3181 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3241 acctgtcgtg ccaggggaat taattctgtc cctcctgttc agctactgac ggggtggtgc
     3301 gtaacggcaa aagcaccgcc ggacatcagc gctagcggag tgtatactgg cttactatgt
     3361 tggcactgat gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg
     3421 gtgcgtcagc agaatatgtg atacaggata tattccgctt cctcgctcac tgactcgcta
     3481 cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg
     3541 gaagatgcca ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc
     3601 ataggctccg cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa
     3661 acccgacagg actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc
     3721 ctgttcctgc ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt
     3781 ccacgcctga cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa
     3841 ccccccgttc agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3901 gaaagacatg caaaagcacc actggcagca gccactggta attgatttag aggagttagt
     3961 cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc
     4021 tccaagccag ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc
     4081 ctgcaaggcg gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca
     4141 agaagatcat cttattaatc agataaaata tttctagcta gagtcgatct tcgccagcag
     4201 ggcgaggatc gtggcatcac cgaaccgcgc cgtgcgcggg tcgtcggtga gccagagttt
     4261 cagcaggccg cccaggcggc ccaggtcgcc attgatgcgg gccagctcgc ggacgtgctc
     4321 atagtccacg acgcccgtga ttttgtagcc ctggccgacg gccagcaggt aggccgacag
     4381 gctcatgccg gccgccgccg ccttttcctc aatcgctctt cgttcgtctg gaaggcagta
     4441 caccttgata ggtgggctgc ccttcctggt tggcttggtt tcatcagcca tccgcttgcc
     4501 ctcatctgtt acgccggcgg tagccggcca gcctcgcaga gcaggattcc cgttgagcac
     4561 cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg ggcctacttc
     4621 acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac gaaccctttg
     4681 gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg ctataatgac
     4741 cccgaagcag ggttatgcag cggaaaagcg ctgcttccct gctgttttgt ggaatatcta
     4801 ccgactggaa acaggcaaat gcaggaaatt actgaactga ggggacaggc gagagacgat
     4861 gccaaagagc tacaccgacg agctggccga gtgggttgaa tcccgcgcgg ccaagaagcg
     4921 ccggcgtgat gaggctgcgg ttgcgttcct ggcggtgagg gcggatgtcg atcgactcta
     4981 gctagaggat cgatcctttt taacccatca catatacctg ccgttcacta ttatttagtg
     5041 aaatgagata ttatgatatt ttctgaattg tgattaaaaa ggcaacttta tgcccatgca
     5101 acagaaacta taaaaaatac agagaatgaa aagaaacaga tagatttttt agttctttag
     5161 gcccgtagtc tgcaaatcct tttatgattt tctatcaaac aaaagaggaa aatagaccag
     5221 ttgcaatcca aacgagagtc taatagaatg aggtcgaaaa gtaaatcgcg cgggtttgtt
     5281 actgataaag caggcaagac ctaaaatgtg taaagggcaa agtgtatact ttggcgtcac
     5341 cccttacata ttttaggtct ttttttattg tgcgtaacta a
//
