LOCUS       62056_22420        7607 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 7607)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            843..952
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             985..1353
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      1974..2362
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     protein_bind    2397..2413
                     /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)."
     CDS             complement(3146..3676)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        4557..4575
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4576..4600
                     /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)."
     CDS             4644..6071
                     /codon_start=1
                     /label=Tn5 transposase
                     /note="transposase from the bacterial Tn5 transposon
                     (Reznikoff, 1993)"
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     terminator      6075..6122
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        6407..6511
                     /label=AmpR promoter
     CDS             6512..7369
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 gaggttttca ccgtcatcac cgaaacgcgc gaggcagcaa ggagatggcg cccaacagtc
       61 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
      121 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
      181 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatccttttt gtccggtgtt
      241 gggttgaagg tgaagccggt cggggccgca gcgggggccg gcttttcagc cttgcccccc
      301 tgcttcggcc gccgtggctc cggcgtcttg ggtgccggcg cgggttccgc agccttggcc
      361 tgcggtgcgg gcacatcggc gggcttggcc ttgatgtgcc gcctggcgtg cgagcggaac
      421 gtctcgtagg agaacttgac cttccccgtt tcccgcatgt gctcccaaat ggtgacgagc
      481 gcatagccgg acgctaacgc cgcctcgaca tccgccctca ccgccaggaa cgcaaccgca
      541 gcctcatcac gccggcgctt cttggccgcg cgggattcaa cccactcggc cagctcgtcg
      601 gtgtagctct ttggcatcgt ctctcgcctg tcccctcagt tcagtaattt cctgcatttg
      661 cctgtttcca gtcggtagat attccacaaa acagcaggga agcagcgctt ttccgctgca
      721 taaccctgct tcggggtcat tatagcgatt ttttcggtat atccatcctt tttcgcacga
      781 tatacaggat tttgccaaag ggttcgtgta gactttcctt ggtgtatcca acggcgtcag
      841 ccgggcagga taggtgaagt aggcccaccc gcgagcgggt gttccttctt cactgtccct
      901 tattcgcacc tggcggtgct caacgggaat cctgctctgc gaggctggcc ggctaccgcc
      961 ggcgtaacag atgagggcaa gcggatggct gatgaaacca agccaaccag gaagggcagc
     1021 ccacctatca aggtgtactg ccttccagac gaacgaagag cgattgagga aaaggcggcg
     1081 gcggccggca tgagcctgtc ggcctacctg ctggccgtcg gccagggcta caaaatcacg
     1141 ggcgtcgtgg actatgagca cgtccgcgag ctggcccgca tcaatggcga cctgggccgc
     1201 ctgggcggcc tgctgaaact ctggctcacc gacgacccgc gcacggcgcg gttcggtgat
     1261 gccacgatcc tcgccctgct ggcgaagatc gaagagaagc aggacgagct tggcaaggtc
     1321 atgatgggcg tggtccgccc gagggcagag ccatgacttt tttagccgct aaaacggccg
     1381 gggggtgcgc gtgattgcca agcacgtccc catgcgctcc atcaagaaga gcgacttcgc
     1441 ggagctggtg aagtacatca ccgacgagca aggcaagacc gagcgcctgg gtcacgtgcg
     1501 cgtcacgaac tgcgaggcaa acaccctgcc cgctgtcatg gccgaggtga tggcgaccca
     1561 gcacggcaac acccgttccg aggccgacaa gacctatcac ctgctggtta gcttccgcgc
     1621 gggagagaag cccgacgcgg agacgttgcg cgcgattgag gaccgcatct gcgctgggct
     1681 tggcttcgcc gagcatcagc gcgtcagtgc cgtgcatcac gacaccgaca acctgcacat
     1741 ccatatcgcc atcaacaaga ttcacccgac ccgaaacacc atccatgagc cgtatcgggc
     1801 ctaccgcgcc ctcgctgacc tctgcgcgac gctcgaacgg gactacgggc ttgagcgtga
     1861 caatcacgaa acgcggcagc gcgtttccga gaaccgcgcg aacgacatgg agcggcacgc
     1921 gggcgtggaa agcctggtcg gctggatccg gccacgatgc gtccggcgta gaggatctga
     1981 agatcagcag ttcaacctgt tgatagtacg tactaagctc tcatgtttca cgtactaagc
     2041 tctcatgttt aacgtactaa gctctcatgt ttaacgaact aaaccctcat ggctaacgta
     2101 ctaagctctc atggctaacg tactaagctc tcatgtttca cgtactaagc tctcatgttt
     2161 gaacaataaa attaatataa atcagcaact taaatagcct ctaaggtttt aagttttata
     2221 agaaaaaaaa gaatatataa ggcttttaaa gcttttaagg tttaacggtt gtggacaaca
     2281 agccagggat gtaacgcact gagaagccct tagagcctct caaagcaatt ttgagtgaca
     2341 caggaacact taacggctga catgggaatt ccacatgtgg aattccacat gtggaattgt
     2401 gagcggataa caatttgtgg aattcccggg agagctcctg actcttatac acaagttcgg
     2461 ccacggccta ggcggccgca gcttgtcgac gaattcccgg ggatccggtg attgattgag
     2521 caagctttat gcttgtaaac cgttttgtga aaaaattttt aaaataaaaa aggggacctc
     2581 tagggtcccc aattaattag taatataatc tattaaaggt cattcaaaag gtcatccacc
     2641 ggatcaattc ccctgctcac acagactggg taccaagctc tcggtaacat caaggcccga
     2701 tccttggagc ccttgccctc ccgacgatga tcgtgccgtg atcgaaatcc agatccttga
     2761 cccacagttg caaaccctca ctgatccgtc ccgtccgggt cgaccaaagc ggccatcgtg
     2821 cctccccact cctgcagttc gggggcatgg atgcgcggat agccgctgct ggtttcctgg
     2881 atgccgacgg atttgcactg ccggtagaac tccgcgaggt cgtccagcct caggcagcag
     2941 ctgaaccaac tcgcgagggg atcgagcccg gggtgggcga agaactccag catgagatcc
     3001 ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca
     3061 tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc
     3121 atttcgaacc ccagagtccc gcttaggtgg cggtacttgg gtcgatatca aagtgcatca
     3181 cttcttcccg tatgcccaac tttgtataga gagccactgc gggatcgtca ccgtaatctg
     3241 cttgcacgta gatcacataa gcaccaagcg cgttggcctc atgcttgagg agattgatga
     3301 gcgcggtggc aatgccctgc ctccggtgct cgccggagac tgcgagatca tagatataga
     3361 tctcactacg cggctgctca aacctgggca gaacgtaagc cgcgagagcg ccaacaaccg
     3421 cttcttggtc gaaggcagca agcgcgatga atgtcttact acggagcaag ttcccgaggt
     3481 aatcggagtc cggctgatgt tgggagtagg tggctacgtc tccgaactca cgaccgaaaa
     3541 gatcaagagc agcccgcatg gatttgactt ggtcagggcc gagcctacat gtgcgaatga
     3601 tgcccatact tgagccacct aactttgttt tagggcgact gccctgctgc gtaacatcgt
     3661 tgctgctgcg taacatgcga aacgatcctc atcctgtctc ttgatcagat cttgatcccc
     3721 tgcgccatca gatccttggc ggcaagaaag ccatccagtt tactttgcag ggcttcccaa
     3781 ccttaccaga gggcgcccca gctggcaatt ccggttcgct tgctgtccat aaaaccgccc
     3841 agtctagcta tcgtcatgta agcccactgc aagctacctc tttctctttg cgcttgcgtt
     3901 ttcccttgtc cagatagccc agtagctgac attcatccgg ggtagcaccg tttctgcgga
     3961 ctggctttct acgtgttccg cttcctttag cagcccttgc gccctgatgc ttgcggcagc
     4021 gtgaagctcg cgcagatcag ttggttgatt ttgtccacta cgtgaaaggc gaatcaccaa
     4081 ggtagtcggc aaataatgtc taacaattcg ttcaagccga cgccgcttcg cggcgcggtt
     4141 aactcaagcg ttagatgcac taagcacata attgctcaca gccaaactat caggtcaagt
     4201 cgcttttatt atttttaagc gtgcataata agccctacac aaaattggga gatatatcat
     4261 gaaaggcgct tttcttgtta tcgcaatagt tgggcgaagt aatcgcaaca tccgcattaa
     4321 caatctagcg agggcttggt aaggtgatcc ggtggatgac cttttgaatg acctttaata
     4381 gattatatta ctaattaatt ggggacccta gaggtcccct tttttatttt aaaaattttt
     4441 tcacaaaacg gtttacaagc ataaagcttg ctcaatcaat caccggatcc ccgggaattc
     4501 ggcctaggtg gccagatctg atcaagagac agaaacgcaa aagaaaatgc cgatattaat
     4561 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt
     4621 aactttaaga aggagatata catatgatta ccagtgcact gcatcgtgcg gcggattggg
     4681 cgaaaagcgt gttttctagt gctgcgctgg gtgatccgcg tcgtaccgcg cgtctggtga
     4741 atgttgcggc gcaactggcc aaatatagcg gcaaaagcat taccattagc agcgaaggca
     4801 gcaaagccat gcaggaaggc gcgtatcgtt ttattcgtaa tccgaacgtg agcgcggaag
     4861 cgattcgtaa agcgggtgcc atgcagaccg tgaaactggc ccaggaattt ccggaactgc
     4921 tggcaattga agataccacc tctctgagct atcgtcatca ggtggcggaa gaactgggca
     4981 aactgggtag cattcaggat aaaagccgtg gttggtgggt gcatagcgtg ctgctgctgg
     5041 aagcgaccac ctttcgtacc gtgggcctgc tgcatcaaga atggtggatg cgtccggatg
     5101 atccggcgga tgcggatgaa aaagaaagcg gcaaatggct ggccgctgct gcaacttcgc
     5161 gtctgagaat gggcagcatg atgagcaacg tgattgcggt gtgcgatcgt gaagcggata
     5221 ttcatgcgta tctgcaagat aaactggccc ataacgaacg ttttgtggtg cgtagcaaac
     5281 atccgcgtaa agatgtggaa agcggcctgt atctgtatga tcacctgaaa aaccagccgg
     5341 aactgggcgg ctatcagatt agcattccgc agaaaggcgt ggtggataaa cgtggcaaac
     5401 gtaaaaaccg tccggcgcgt aaagcgagcc tgagcctgcg tagcggccgt attaccctga
     5461 aacagggcaa cattaccctg aacgcggtgc tggccgaaga aattaatccg ccgaaaggcg
     5521 aaaccccgct gaaatggctg ctgctgacca gcgagccggt ggaaagtctg gcccaagcgc
     5581 tgcgtgtgat tgatatttat acccatcgtt ggcgcattga agaatttcac aaagcgtgga
     5641 aaacgggtgc gggtgcggaa cgtcagcgta tggaagaacc ggataacctg gaacgtatgg
     5701 tgagcattct gagctttgtg gcggtgcgtc tgctgcaact gcgtgaatct tttactccgc
     5761 cgcaagcact gcgtgcgcag ggcctgctga aagaagcgga acacgttgaa agccagagcg
     5821 cggaaaccgt gctgaccccg gatgaatgcc aactgctggg ctatctggat aaaggcaaac
     5881 gcaaacgcaa agaaaaagcg ggcagcctgc aatgggcgta tatggcgatt gcgcgtctgg
     5941 gcggctttat ggatagcaaa cgtaccggca ttgcgagctg gggtgcgctg tgggaaggtt
     6001 gggaagcgct gcaaagcaaa ctggatggct ttctggccgc gaaagacctg atggcgcagg
     6061 gcattaaaat ctaactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     6121 tgacgcgtaa aacagccagc gctggcgcga tttagccccg acatagcccc actgttcgtc
     6181 catttccgcg cagacgatga cgtcactgcc cggctgtatg cgcgaggtta ccgactgcgg
     6241 cctgagtttt ttaagtgacg taaaatcgtg ttgaggccaa cgcccataat gcgggctgtt
     6301 gcccggcatc caacgccatt catggccata tcaatgattt tctggtgcgt accgggttga
     6361 gaagcggtgt aagtgaactg caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     6421 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6481 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     6541 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6601 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     6661 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     6721 taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg
     6781 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6841 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6901 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6961 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     7021 cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa
     7081 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     7141 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     7201 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     7261 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     7321 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     7381 ccaagtttac tcatatatac tttagattga tttatggtgc actctcagta caatctgctc
     7441 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     7501 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     7561 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtca
//
