LOCUS       62056_13350        6666 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 6666)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(14..889)
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        916..1200
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     CDS             1256..1669
                     /codon_start=1
                     /label=Gam
                     /note="inhibitor of the host RecBCD nuclease in the lambda 
                     Red system"
                     /translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
                     IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
                     AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
     CDS             1678..2460
                     /codon_start=1
                     /label=Beta
                     /note="single-stranded DNA binding recombinase in the 
                     lambda Red system"
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
     CDS             2460..3137
                     /codon_start=1
                     /label=Exo
                     /note="5' to 3' double-stranded DNA exonuclease in the 
                     lambda
                     Red system"
                     /translation="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKPR
                     SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
                     IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
                     VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
                     EQWR"
     terminator      3141..3385
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     CDS             complement(3511..4458)
                     /codon_start=1
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      complement(4506..4728)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     promoter        5374..5402
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             5450..6637
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
ORIGIN
        1 catcgattta ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac
       61 ggaactcgct cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat
      121 cgtcaaaacc aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca
      181 gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct
      241 ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga
      301 tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat
      361 tatccatcgg tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct
      421 caagcagatt tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga
      481 tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg
      541 tattggcaaa tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt
      601 aaacccactg gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc
      661 ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca
      721 ccaccccctg accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt
      781 cgataaaaaa atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg
      841 cattaaacga gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac
      901 tcccgccatt cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg
      961 tcttttactg gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt
     1021 aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca
     1081 gaaaagtcca cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat
     1141 ccataagatt agcggatcct acctgacgct ttttatcgca actctctact gtttctccat
     1201 acccgttttt ttgggaattc gagctcggta ccctcgagta aggaggttat aaaaaatgga
     1261 tattaatact gaaactgaga tcaagcaaaa gcattcacta accccctttc ctgttttcct
     1321 aatcagcccg gcatttcgcg ggcgatattt tcacagctat ttcaggagtt cagccatgaa
     1381 cgcttattac attcaggatc gtcttgaggc tcagagctgg gcgcgtcact accagcagct
     1441 cgcccgtgaa gagaaagagg cagaactggc agacgacatg gaaaaaggcc tgccccagca
     1501 cctgtttgaa tcgctatgca tcgatcattt gcaacgccac ggggccagca aaaaatccat
     1561 tacccgtgcg tttgatgacg atgttgagtt tcaggagcgc atggcagaac acatccggta
     1621 catggttgaa accattgctc accaccaggt tgatattgat tcagaggtat aaaacgaatg
     1681 agtactgcac tcgcaacgct ggctgggaag ctggctgaac gtgtcggcat ggattctgtc
     1741 gacccacagg aactgatcac cactcttcgc cagacggcat ttaaaggtga tgccagcgat
     1801 gcgcagttca tcgcattact gatcgttgcc aaccagtacg gccttaatcc gtggacgaaa
     1861 gaaatttacg cctttcctga taagcagaat ggcatcgttc cggtggtggg cgttgatggc
     1921 tggtcccgca tcatcaatga aaaccagcag tttgatggca tggactttga gcaggacaat
     1981 gaatcctgta catgccggat ttaccgcaag gaccgtaatc atccgatctg cgttaccgaa
     2041 tggatggatg aatgccgccg cgaaccattc aaaactcgcg aaggcagaga aatcacgggg
     2101 ccgtggcagt cgcatcccaa acggatgtta cgtcataaag ccatgattca gtgtgcccgt
     2161 ctggccttcg gatttgctgg tatctatgac aaggatgaag ccgagcgcat tgtcgaaaat
     2221 actgcataca ctgcagaacg tcagccggaa cgcgacatca ctccggttaa cgatgaaacc
     2281 atgcaggaga ttaacactct gctgatcgcc ctggataaaa catgggatga cgacttattg
     2341 ccgctctgtt cccagatatt tcgccgcgac attcgtgcat cgtcagaact gacacaggcc
     2401 gaagcagtaa aagctcttgg attcctgaaa cagaaagccg cagagcagaa ggtggcagca
     2461 tgacaccgga cattatcctg cagcgtaccg ggatcgatgt gagagctgtc gaacaggggg
     2521 atgatgcgtg gcacaaatta cggctcggcg tcatcaccgc ttcagaagtt cacaacgtga
     2581 tagcaaaacc ccgctccgga aagaagtggc ctgacatgaa aatgtcctac ttccacaccc
     2641 tgcttgctga ggtttgcacc ggtgtggctc cggaagttaa cgctaaagca ctggcctggg
     2701 gaaaacagta cgagaacgac gccagaaccc tgtttgaatt cacttccggc gtgaatgtta
     2761 ctgaatcccc gatcatctat cgcgacgaaa gtatgcgtac cgcctgctct cccgatggtt
     2821 tatgcagtga cggcaacggc cttgaactga aatgcccgtt tacctcccgg gatttcatga
     2881 agttccggct cggtggtttc gaggccataa agtcagctta catggcccag gtgcagtaca
     2941 gcatgtgggt gacgcgaaaa aatgcctggt actttgccaa ctatgacccg cgtatgaagc
     3001 gtgaaggcct gcattatgtc gtgattgagc gggatgaaaa gtacatggcg agttttgacg
     3061 agatcgtgcc ggagttcatc gaaaaaatgg acgaggcact ggctgaaatt ggttttgtat
     3121 ttggggagca atggcgatga cgcatcctca cgataatatc cgggtaggcg caatcacttt
     3181 cgtctactcc gttacaaagc gaggctgggt atttcccggc ctttctgtta tccgaaatcc
     3241 actgaaagca cagcggctgg ctgaggagat aaataataaa cgaggggctg tatgcacaaa
     3301 gcatcttctg ttgagttaag aacgagtatc gagatggcac atagccttgc tcaaattgga
     3361 atcaggtttg tgccaatacc agtagaaaca gacgaagaat ccatgggtat ggacagtttt
     3421 ccctttgata tgtaacggtg aacagttgtt ctacttttgt ttgttagtct tgatgcttca
     3481 ctgatagata caagagccat aagaacctca gatccttccg tatttagcca gtatgttctc
     3541 tagtgtggtt cgttgttttt gcgtgagcca tgagaacgaa ccattgagat catacttact
     3601 ttgcatgtca ctcaaaaatt ttgcctcaaa actggtgagc tgaatttttg cagttaaagc
     3661 atcgtgtagt gtttttctta gtccgttacg taggtaggaa tctgatgtaa tggttgttgg
     3721 tattttgtca ccattcattt ttatctggtt gttctcaagt tcggttacga gatccatttg
     3781 tctatctagt tcaacttgga aaatcaacgt atcagtcggg cggcctcgct tatcaaccac
     3841 caatttcata ttgctgtaag tgtttaaatc tttacttatt ggtttcaaaa cccattggtt
     3901 aagcctttta aactcatggt agttattttc aagcattaac atgaacttaa attcatcaag
     3961 gctaatctct atatttgcct tgtgagtttt cttttgtgtt agttctttta ataaccactc
     4021 ataaatcctc atagagtatt tgttttcaaa agacttaaca tgttccagat tatattttat
     4081 gaattttttt aactggaaaa gataaggcaa tatctcttca ctaaaaacta attctaattt
     4141 ttcgcttgag aacttggcat agtttgtcca ctggaaaatc tcaaagcctt taaccaaagg
     4201 attcctgatt tccacagttc tcgtcatcag ctctctggtt gctttagcta atacaccata
     4261 agcattttcc ctactgatgt tcatcatctg agcgtattgg ttataagtga acgataccgt
     4321 ccgttctttc cttgtagggt tttcaatcgt ggggttgagt agtgccacac agcataaaat
     4381 tagcttggtt tcatgctccg ttaagtcata gcgactaatc gctagttcat ttgctttgaa
     4441 aacaactaat tcagacatac atctcaattg gtctaggtga ttttaatcac tataccaatt
     4501 gagatgggct agtcaatgat aattactagt ccttttcctt tgagttgtgg gtatctgtaa
     4561 attctgctag acctttgctg gaaaacttgt aaattctgct agaccctctg taaattccgc
     4621 tagacctttg tgtgtttttt ttgtttatat tcaagtggtt ataatttata gaataaagaa
     4681 agaataaaaa aagataaaaa gaatagatcc cagccctgtg tataactcac tactttagtc
     4741 agttccgcag tattacaaaa ggatgtcgca aacgctgttt gctcctctac aaaacagacc
     4801 ttaaaaccct aaaggcttaa gtagcaccct cgcaagctcg gttgcggccg caatcgggca
     4861 aatcgctgaa tattcctttt gtctccgacc atcaggcacc tgagtcgctg tctttttcgt
     4921 gacattcagt tcgctgcgct cacggctctg gcagtgaatg ggggtaaatg gcactacagg
     4981 cgccttttat ggattcatgc aaggaaacta cccataatac aagaaaagcc cgtcacgggc
     5041 ttctcagggc gttttatggc gggtctgcta tgtggtgcta tctgactttt tgctgttcag
     5101 cagttcctgc cctctgattt tccagtctga ccacttcgga ttatcccgtg acaggtcatt
     5161 cagactggct aatgcaccca gtaaggcagc ggtatcatca acggggtctg acgctcagtg
     5221 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5281 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5341 gtctgacagt ttcgtcttca agaattctca tgtttgacag cttatcatcg ataagcttta
     5401 atgcggtagt ttatcacagt taaattgcta acgcagtcag gcaccgtgta tgaaatctaa
     5461 caatgcgctc atcgtcatcc tcggcaccgt caccctggat gctgtaggca taggcttggt
     5521 tatgccggta ctgccgggcc tcttgcggga tatcgtccat tccgacagca tcgccagtca
     5581 ctatggcgtg ctgctagcgc tatatgcgtt gatgcaattt ctatgcgcac ccgttctcgg
     5641 agcactgtcc gaccgctttg gccgccgccc agtcctgctc gcttcgctac ttggagccac
     5701 tatcgactac gcgatcatgg cgaccacacc cgtcctgtgg atcctctacg ccggacgcat
     5761 cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac
     5821 cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat
     5881 ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct
     5941 tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc
     6001 gcataaggga gagcgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg
     6061 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
     6121 cgtaggacag gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag
     6181 cgcgacgatg atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc
     6241 cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat
     6301 ggcggccgac gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt
     6361 ccccattatg attcttctcg cttccggcgg catcgggatg cccgcgttgc aggccatgct
     6421 gtccaggcag gtagatgacg accatcaggg acagcttcaa ggatcgctcg cggctcttac
     6481 cagcctaact tcgatcactg gaccgctgat cgtcacggcg atttatgccg cctcggcgag
     6541 cacatggaac gggttggcat ggattgtagg cgccgcccta taccttgtct gcctccccgc
     6601 gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga cacatttccc cgaaaagtgc
     6661 cacctg
//
