LOCUS       62056_22400        8335 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 8335)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8335
                     /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             2480..3196
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(3874..4404)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        5285..5303
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5304..5328
                     /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             5372..6799
                     /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      6803..6850
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        7135..7239
                     /label=AmpR promoter
     CDS             7240..8097
                     /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 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     2521 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     2581 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     2641 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     2701 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2761 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2821 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2881 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     2941 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     3001 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     3061 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     3121 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     3181 ggacgagctg tacaagtagg cggccgcagc ttgtcgacga attcccgggg atccggtgat
     3241 tgattgagca agctttatgc ttgtaaaccg ttttgtgaaa aaatttttaa aataaaaaag
     3301 gggacctcta gggtccccaa ttaattagta atataatcta ttaaaggtca ttcaaaaggt
     3361 catccaccgg atcaattccc ctgctcacac agactgggta ccaagctctc ggtaacatca
     3421 aggcccgatc cttggagccc ttgccctccc gacgatgatc gtgccgtgat cgaaatccag
     3481 atccttgacc cacagttgca aaccctcact gatccgtccc gtccgggtcg accaaagcgg
     3541 ccatcgtgcc tccccactcc tgcagttcgg gggcatggat gcgcggatag ccgctgctgg
     3601 tttcctggat gccgacggat ttgcactgcc ggtagaactc cgcgaggtcg tccagcctca
     3661 ggcagcagct gaaccaactc gcgaggggat cgagcccggg gtgggcgaag aactccagca
     3721 tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca
     3781 acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt
     3841 ggtcggtcat ttcgaacccc agagtcccgc ttaggtggcg gtacttgggt cgatatcaaa
     3901 gtgcatcact tcttcccgta tgcccaactt tgtatagaga gccactgcgg gatcgtcacc
     3961 gtaatctgct tgcacgtaga tcacataagc accaagcgcg ttggcctcat gcttgaggag
     4021 attgatgagc gcggtggcaa tgccctgcct ccggtgctcg ccggagactg cgagatcata
     4081 gatatagatc tcactacgcg gctgctcaaa cctgggcaga acgtaagccg cgagagcgcc
     4141 aacaaccgct tcttggtcga aggcagcaag cgcgatgaat gtcttactac ggagcaagtt
     4201 cccgaggtaa tcggagtccg gctgatgttg ggagtaggtg gctacgtctc cgaactcacg
     4261 accgaaaaga tcaagagcag cccgcatgga tttgacttgg tcagggccga gcctacatgt
     4321 gcgaatgatg cccatacttg agccacctaa ctttgtttta gggcgactgc cctgctgcgt
     4381 aacatcgttg ctgctgcgta acatgcgaaa cgatcctcat cctgtctctt gatcagatct
     4441 tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta ctttgcaggg
     4501 cttcccaacc ttaccagagg gcgccccagc tggcaattcc ggttcgcttg ctgtccataa
     4561 aaccgcccag tctagctatc gtcatgtaag cccactgcaa gctacctctt tctctttgcg
     4621 cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatccgggg tagcaccgtt
     4681 tctgcggact ggctttctac gtgttccgct tcctttagca gcccttgcgc cctgatgctt
     4741 gcggcagcgt gaagctcgcg cagatcagtt ggttgatttt gtccactacg tgaaaggcga
     4801 atcaccaagg tagtcggcaa ataatgtcta acaattcgtt caagccgacg ccgcttcgcg
     4861 gcgcggttaa ctcaagcgtt agatgcacta agcacataat tgctcacagc caaactatca
     4921 ggtcaagtcg cttttattat ttttaagcgt gcataataag ccctacacaa aattgggaga
     4981 tatatcatga aaggcgcttt tcttgttatc gcaatagttg ggcgaagtaa tcgcaacatc
     5041 cgcattaaca atctagcgag ggcttggtaa ggtgatccgg tggatgacct tttgaatgac
     5101 ctttaataga ttatattact aattaattgg ggaccctaga ggtccccttt tttattttaa
     5161 aaattttttc acaaaacggt ttacaagcat aaagcttgct caatcaatca ccggatcccc
     5221 gggaattcgg cctaggtggc cagatctgat caagagacag aaacgcaaaa gaaaatgccg
     5281 atattaatac gactcactat aggggaattg tgagcggata acaattcccc tctagaaata
     5341 attttgttaa ctttaagaag gagatataca tatgattacc agtgcactgc atcgtgcggc
     5401 ggattgggcg aaaagcgtgt tttctagtgc tgcgctgggt gatccgcgtc gtaccgcgcg
     5461 tctggtgaat gttgcggcgc aactggccaa atatagcggc aaaagcatta ccattagcag
     5521 cgaaggcagc aaagccatgc aggaaggcgc gtatcgtttt attcgtaatc cgaacgtgag
     5581 cgcggaagcg attcgtaaag cgggtgccat gcagaccgtg aaactggccc aggaatttcc
     5641 ggaactgctg gcaattgaag ataccacctc tctgagctat cgtcatcagg tggcggaaga
     5701 actgggcaaa ctgggtagca ttcaggataa aagccgtggt tggtgggtgc atagcgtgct
     5761 gctgctggaa gcgaccacct ttcgtaccgt gggcctgctg catcaagaat ggtggatgcg
     5821 tccggatgat ccggcggatg cggatgaaaa agaaagcggc aaatggctgg ccgctgctgc
     5881 aacttcgcgt ctgagaatgg gcagcatgat gagcaacgtg attgcggtgt gcgatcgtga
     5941 agcggatatt catgcgtatc tgcaagataa actggcccat aacgaacgtt ttgtggtgcg
     6001 tagcaaacat ccgcgtaaag atgtggaaag cggcctgtat ctgtatgatc acctgaaaaa
     6061 ccagccggaa ctgggcggct atcagattag cattccgcag aaaggcgtgg tggataaacg
     6121 tggcaaacgt aaaaaccgtc cggcgcgtaa agcgagcctg agcctgcgta gcggccgtat
     6181 taccctgaaa cagggcaaca ttaccctgaa cgcggtgctg gccgaagaaa ttaatccgcc
     6241 gaaaggcgaa accccgctga aatggctgct gctgaccagc gagccggtgg aaagtctggc
     6301 ccaagcgctg cgtgtgattg atatttatac ccatcgttgg cgcattgaag aatttcacaa
     6361 agcgtggaaa acgggtgcgg gtgcggaacg tcagcgtatg gaagaaccgg ataacctgga
     6421 acgtatggtg agcattctga gctttgtggc ggtgcgtctg ctgcaactgc gtgaatcttt
     6481 tactccgccg caagcactgc gtgcgcaggg cctgctgaaa gaagcggaac acgttgaaag
     6541 ccagagcgcg gaaaccgtgc tgaccccgga tgaatgccaa ctgctgggct atctggataa
     6601 aggcaaacgc aaacgcaaag aaaaagcggg cagcctgcaa tgggcgtata tggcgattgc
     6661 gcgtctgggc ggctttatgg atagcaaacg taccggcatt gcgagctggg gtgcgctgtg
     6721 ggaaggttgg gaagcgctgc aaagcaaact ggatggcttt ctggccgcga aagacctgat
     6781 ggcgcagggc attaaaatct aactagcata accccttggg gcctctaaac gggtcttgag
     6841 gggttttttg acgcgtaaaa cagccagcgc tggcgcgatt tagccccgac atagccccac
     6901 tgttcgtcca tttccgcgca gacgatgacg tcactgcccg gctgtatgcg cgaggttacc
     6961 gactgcggcc tgagtttttt aagtgacgta aaatcgtgtt gaggccaacg cccataatgc
     7021 gggctgttgc ccggcatcca acgccattca tggccatatc aatgattttc tggtgcgtac
     7081 cgggttgaga agcggtgtaa gtgaactgca ggtggcactt ttcggggaaa tgtgcgcgga
     7141 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     7201 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     7261 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     7321 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     7381 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     7441 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     7501 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     7561 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     7621 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     7681 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     7741 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     7801 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     7861 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     7921 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     7981 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     8041 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     8101 ctgtcagacc aagtttactc atatatactt tagattgatt tatggtgcac tctcagtaca
     8161 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     8221 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     8281 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtca
//
