LOCUS       62056_13525        5681 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 5681)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        118..220
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             221..877
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     rep_origin      1124..1579
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1683..2128
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             2129..3547
                     /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(3951..4898)
                     /codon_start=1
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      complement(4946..5168)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 ccctttcgtc ttcgaataaa tacctgtgac ggaagatcac ttcgcagaat aaataaatcc
       61 tggtgtccct gttgataccg ggaagccctg ggccaacttt tggcgaaaat gagacgttga
      121 tcggcacgta agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt
      181 ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg
      241 atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc
      301 agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac
      361 cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat
      421 gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag
      481 tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag
      541 tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta
      601 cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc
      661 caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt
      721 cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
      781 ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga
      841 attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg
      901 gtgcccttaa acgcctggtg ctacgcctga ataagtgata ataagcggat gaatggcaga
      961 aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt
     1021 atgtctattg ctggtttaat cggtacccgg ggatcgcggc cgcggaccgg atcctctaga
     1081 gcggccgcga tcctctagag tcgaccggag aatggcgaat gggacgcgcc ctgtagcggc
     1141 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1201 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1261 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     1321 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     1381 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     1441 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     1501 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     1561 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1621 gtttattttt ctaaatacat tcaaatatgt atccgctcat nncangatcc tttttaaccc
     1681 atcacatata cctgccgttc actattattt agtgaaatga gatattatga tattttctga
     1741 attgtgatta aaaaggcaac tttatgccca tgcaacagaa actataaaaa atacagagaa
     1801 tgaaaagaaa cagatagatt ttttagttct ttaggcccgt agtctgcaaa tccttttatg
     1861 attttctatc aaacaaaaga ggaaaataga ccagttgcaa tccaaacgag agtctaatag
     1921 aatgaggtcg aaaagtaaat cgcgcgggtt tgttactgat aaagcaggca agacctaaaa
     1981 tgtgtaaagg gcaaagtgta tactttggcg tcacccctta catattttag gtcttttttt
     2041 attgtgcgta actaacttgc catcttcaaa caggagggct ggaagaagca gaccgctaac
     2101 acagtacata aaaaaggaga catgaacgat gaacatcaaa aagtttgcaa aacaagcaac
     2161 agtattaacc tttactaccg cactgctggc aggaggcgca actcaagcgt ttgcgaaaga
     2221 aacgaaccaa aagccatata aggaaacata cggcatttcc catattacac gccatgatat
     2281 gctgcaaatc cctgaacagc aaaaaaatga aaaatatcaa gttcctgagt tcgattcgtc
     2341 cacaattaaa aatatctctt ctgcaaaagg cctggacgtt tgggacagct ggccattaca
     2401 aaacgctgac ggcactgtcg caaactatca cggctaccac atcgtctttg cattagccgg
     2461 agatcctaaa aatgcggatg acacatcgat ttacatgttc tatcaaaaag tcggcgaaac
     2521 ttctattgac agctggaaaa acgctggccg cgtctttaaa gacagcgaca aattcgatgc
     2581 aaatgattct atcctaaaag accaaacaca agaatggtca ggttcagcca catttacatc
     2641 tgacggaaaa atccgtttat tctacactga tttctccggt aaacattacg gcaaacaaac
     2701 actgacaact gcacaagtta acgtatcagc atcagacagc tctttgaaca tcaacggtgt
     2761 agaggattat aaatcaatct ttgacggtga cggaaaaacg tatcaaaatg tacagcagtt
     2821 catcgatgaa ggcaactaca gctcaggcga caaccatacg ctgagagatc ctcactacgt
     2881 agaagataaa ggccacaaat acttagtatt tgaagcaaac actggaactg aagatggcta
     2941 ccaaggcgaa gaatctttat ttaacaaagc atactatggc aaaagcacat cattcttccg
     3001 tcaagaaagt caaaaacttc tgcaaagcga taaaaaacgc acggctgagt tagcaaacgg
     3061 cgctctcggt atgattgagc taaacgatga ttacacactg aaaaaagtga tgaaaccgct
     3121 gattgcatct aacacagtaa cagatgaaat tgaacgcgcg aacgtcttta aaatgaacgg
     3181 caaatggtac ctgttcactg actcccgcgg atcaaaaatg acgattgacg gcattacgtc
     3241 taacgatatt tacatgcttg gttatgtttc taattcttta actggcccat acaagccgct
     3301 gaacaaaact ggccttgtgt taaaaatgga tcttgatcct aacgatgtaa cctttactta
     3361 ctcacacttc gctgtacctc aagcgaaagg aaacaatgtc gtgattacaa gctatatgac
     3421 aaacagagga ttctacgcag acaaacaatc aacgtttgcg ccaagcttcc tgctgaacat
     3481 caaaggcaag aaaacatctg ttgtcaaaga cagcatcctt gagcaaggac aattaacagt
     3541 taacaaataa aaacgcaaaa gaaaatgccg atnnccggtt tattgactac cggaagcagt
     3601 gtgaccgtgt gcttctcaaa tgcctcaggc tgtctatgtg tgactgttga gctgtaacaa
     3661 gttgtctcag gtgttcaatt tcatgttcta gttgctttgt tttactggtt tcacctgttc
     3721 tattaggtgt tacatgctgt tcatctgtta cattgtcgat ctgttcatgg tgaacagctt
     3781 taaatgcacc aaaaactcgt aaaagctctg atgtatctat cttttttaca ccgttttcat
     3841 ctgtgcatat ggacagtttt ccctttgata tctaacggtg aacagttgtt ctacttttgt
     3901 ttgttagtct tgatgcttca ctgatagata caagagccat aagaacctca gatccttccg
     3961 tatttagcca gtatgttctc tagtgtggtt cgttgttttt gcgtgagcca tgagaacgaa
     4021 ccattgagat catgcttact ttgcatgtca ctcaaaaatt ttgcctcaaa actggtgagc
     4081 tgaatttttg cagttaaagc atcgtgtagt gtttttctta gtccgttacg taggtaggaa
     4141 tctgatgtaa tggttgttgg tattttgtca ccattcattt ttatctggtt gttctcaagt
     4201 tcggttacga gatccatttg tctatctagt tcaacttgga aaatcaacgt atcagtcggg
     4261 cggcctcgct tatcaaccac caatttcata ttgctgtaag tgtttaaatc tttacttatt
     4321 ggtttcaaaa cccattggtt aagcctttta aactcatggt agttattttc aagcattaac
     4381 atgaacttaa attcatcaag gctaatctct atatttgcct tgtgagtttt cttttgtgtt
     4441 agttctttta ataaccactc ataaatcctc atagagtatt tgttttcaaa agacttaaca
     4501 tgttccagat tatattttat gaattttttt aactggaaaa gataaggcaa tatctcttca
     4561 ctaaaaacta attctaattt ttcgcttgag aacttggcat agtttgtcca ctggaaaatc
     4621 tcaaagcctt taaccaaagg attcctgatt tccacagttc tcgtcatcag ctctctggtt
     4681 gctttagcta atacaccata agcattttcc ctactgatgt tcatcatctg agcgtattgg
     4741 ttataagtga acgataccgt ccgttctttc cttgtagggt tttcaatcgt ggggttgagt
     4801 agtgccacac agcataaaat tagcttggtt tcatgctccg ttaagtcata gcgactaatc
     4861 gctagttcat ttgctttgaa aacaactaat tcagacatac atctcaattg gtctaggtga
     4921 ttttaatcac tataccaatt gagatgggct agtcaatgat aattactagt ccttttcctt
     4981 tgagttgtgg gtatctgtaa attctgctag acctttgctg gaaaacttgt aaattctgct
     5041 agaccctctg taaattccgc tagacctttg tgtgtttttt ttgtttatat tcaagtggtt
     5101 ataatttata gaataaagaa agaataaaaa aagataaaaa gaatagatcc cagccctgtg
     5161 tataactcac tactttagtc agttccgcag tattacaaaa ggatgtcgca aacgctgttt
     5221 gctcctctac aaaacagacc ttaaaaccct aaaggcttaa gtagcaccct cgcaagctcg
     5281 ggcaaatcgc tgaatattcc ttttgtctcc gaccatcagg cacctgagtc gctgtctttt
     5341 tcgtgacatt cagttcgctg cgctcacggc tctggcagtg aatgggggta aatggcacta
     5401 caggcgcctt ttatggattc atgcaaggaa actacccata atacaagaaa agcccgtcac
     5461 gggcttctca gggcgtttta tggcgggtct gctatgtggt gctatctgac tttttgctgt
     5521 tcagcagttc ctgccctctg attttccagt ctgaccactt cggattatcc cgtgacaggt
     5581 cattcagact ggctaatgca cccagtaagg cagcggtatc atcaacaggc ttacccgtct
     5641 tactgtcngg atcgacgctc tcccttatgc gactcctgca t
//
