LOCUS       62056_22410        8371 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 8371)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8371
                     /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)."
     promoter        2480..2508
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             2516..3232
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(3910..4440)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        5321..5339
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5340..5364
                     /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             5408..6835
                     /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      6839..6886
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        7171..7275
                     /label=AmpR promoter
     CDS             7276..8133
                     /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 ggcggccgct tgacaattaa tcatcggctc gtataatgtg tggaaatggt
     2521 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     2581 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     2641 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     2701 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     2761 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     2821 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     2881 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     2941 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     3001 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     3061 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     3121 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     3181 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaggcggc
     3241 cgcagcttgt cgacgaattc ccggggatcc ggtgattgat tgagcaagct ttatgcttgt
     3301 aaaccgtttt gtgaaaaaat ttttaaaata aaaaagggga cctctagggt ccccaattaa
     3361 ttagtaatat aatctattaa aggtcattca aaaggtcatc caccggatca attcccctgc
     3421 tcacacagac tgggtaccaa gctctcggta acatcaaggc ccgatccttg gagcccttgc
     3481 cctcccgacg atgatcgtgc cgtgatcgaa atccagatcc ttgacccaca gttgcaaacc
     3541 ctcactgatc cgtcccgtcc gggtcgacca aagcggccat cgtgcctccc cactcctgca
     3601 gttcgggggc atggatgcgc ggatagccgc tgctggtttc ctggatgccg acggatttgc
     3661 actgccggta gaactccgcg aggtcgtcca gcctcaggca gcagctgaac caactcgcga
     3721 ggggatcgag cccggggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca
     3781 tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg
     3841 aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag
     3901 tcccgcttag gtggcggtac ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc
     3961 caactttgta tagagagcca ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac
     4021 ataagcacca agcgcgttgg cctcatgctt gaggagattg atgagcgcgg tggcaatgcc
     4081 ctgcctccgg tgctcgccgg agactgcgag atcatagata tagatctcac tacgcggctg
     4141 ctcaaacctg ggcagaacgt aagccgcgag agcgccaaca accgcttctt ggtcgaaggc
     4201 agcaagcgcg atgaatgtct tactacggag caagttcccg aggtaatcgg agtccggctg
     4261 atgttgggag taggtggcta cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg
     4321 catggatttg acttggtcag ggccgagcct acatgtgcga atgatgccca tacttgagcc
     4381 acctaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tgcgtaacat
     4441 gcgaaacgat cctcatcctg tctcttgatc agatcttgat cccctgcgcc atcagatcct
     4501 tggcggcaag aaagccatcc agtttacttt gcagggcttc ccaaccttac cagagggcgc
     4561 cccagctggc aattccggtt cgcttgctgt ccataaaacc gcccagtcta gctatcgtca
     4621 tgtaagccca ctgcaagcta cctctttctc tttgcgcttg cgttttccct tgtccagata
     4681 gcccagtagc tgacattcat ccggggtagc accgtttctg cggactggct ttctacgtgt
     4741 tccgcttcct ttagcagccc ttgcgccctg atgcttgcgg cagcgtgaag ctcgcgcaga
     4801 tcagttggtt gattttgtcc actacgtgaa aggcgaatca ccaaggtagt cggcaaataa
     4861 tgtctaacaa ttcgttcaag ccgacgccgc ttcgcggcgc ggttaactca agcgttagat
     4921 gcactaagca cataattgct cacagccaaa ctatcaggtc aagtcgcttt tattattttt
     4981 aagcgtgcat aataagccct acacaaaatt gggagatata tcatgaaagg cgcttttctt
     5041 gttatcgcaa tagttgggcg aagtaatcgc aacatccgca ttaacaatct agcgagggct
     5101 tggtaaggtg atccggtgga tgaccttttg aatgaccttt aatagattat attactaatt
     5161 aattggggac cctagaggtc ccctttttta ttttaaaaat tttttcacaa aacggtttac
     5221 aagcataaag cttgctcaat caatcaccgg atccccggga attcggccta ggtggccaga
     5281 tctgatcaag agacagaaac gcaaaagaaa atgccgatat taatacgact cactataggg
     5341 gaattgtgag cggataacaa ttcccctcta gaaataattt tgttaacttt aagaaggaga
     5401 tatacatatg attaccagtg cactgcatcg tgcggcggat tgggcgaaaa gcgtgttttc
     5461 tagtgctgcg ctgggtgatc cgcgtcgtac cgcgcgtctg gtgaatgttg cggcgcaact
     5521 ggccaaatat agcggcaaaa gcattaccat tagcagcgaa ggcagcaaag ccatgcagga
     5581 aggcgcgtat cgttttattc gtaatccgaa cgtgagcgcg gaagcgattc gtaaagcggg
     5641 tgccatgcag accgtgaaac tggcccagga atttccggaa ctgctggcaa ttgaagatac
     5701 cacctctctg agctatcgtc atcaggtggc ggaagaactg ggcaaactgg gtagcattca
     5761 ggataaaagc cgtggttggt gggtgcatag cgtgctgctg ctggaagcga ccacctttcg
     5821 taccgtgggc ctgctgcatc aagaatggtg gatgcgtccg gatgatccgg cggatgcgga
     5881 tgaaaaagaa agcggcaaat ggctggccgc tgctgcaact tcgcgtctga gaatgggcag
     5941 catgatgagc aacgtgattg cggtgtgcga tcgtgaagcg gatattcatg cgtatctgca
     6001 agataaactg gcccataacg aacgttttgt ggtgcgtagc aaacatccgc gtaaagatgt
     6061 ggaaagcggc ctgtatctgt atgatcacct gaaaaaccag ccggaactgg gcggctatca
     6121 gattagcatt ccgcagaaag gcgtggtgga taaacgtggc aaacgtaaaa accgtccggc
     6181 gcgtaaagcg agcctgagcc tgcgtagcgg ccgtattacc ctgaaacagg gcaacattac
     6241 cctgaacgcg gtgctggccg aagaaattaa tccgccgaaa ggcgaaaccc cgctgaaatg
     6301 gctgctgctg accagcgagc cggtggaaag tctggcccaa gcgctgcgtg tgattgatat
     6361 ttatacccat cgttggcgca ttgaagaatt tcacaaagcg tggaaaacgg gtgcgggtgc
     6421 ggaacgtcag cgtatggaag aaccggataa cctggaacgt atggtgagca ttctgagctt
     6481 tgtggcggtg cgtctgctgc aactgcgtga atcttttact ccgccgcaag cactgcgtgc
     6541 gcagggcctg ctgaaagaag cggaacacgt tgaaagccag agcgcggaaa ccgtgctgac
     6601 cccggatgaa tgccaactgc tgggctatct ggataaaggc aaacgcaaac gcaaagaaaa
     6661 agcgggcagc ctgcaatggg cgtatatggc gattgcgcgt ctgggcggct ttatggatag
     6721 caaacgtacc ggcattgcga gctggggtgc gctgtgggaa ggttgggaag cgctgcaaag
     6781 caaactggat ggctttctgg ccgcgaaaga cctgatggcg cagggcatta aaatctaact
     6841 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgacgc gtaaaacagc
     6901 cagcgctggc gcgatttagc cccgacatag ccccactgtt cgtccatttc cgcgcagacg
     6961 atgacgtcac tgcccggctg tatgcgcgag gttaccgact gcggcctgag ttttttaagt
     7021 gacgtaaaat cgtgttgagg ccaacgccca taatgcgggc tgttgcccgg catccaacgc
     7081 cattcatggc catatcaatg attttctggt gcgtaccggg ttgagaagcg gtgtaagtga
     7141 actgcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     7201 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     7261 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     7321 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     7381 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     7441 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     7501 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     7561 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     7621 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     7681 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     7741 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     7801 cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa
     7861 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     7921 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     7981 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     8041 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     8101 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     8161 atactttaga ttgatttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     8221 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     8281 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     8341 ctgtgaccgt ctccgggagc tgcatgtgtc a
//
