LOCUS       62056_22430        7868 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 7868)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(109..133)
                     /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        complement(134..152)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1033..1824
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    complement(2557..2573)
                     /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)."
     rep_origin      complement(2608..2996)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(3617..3985)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(4018..4127)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(5208..6065)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6066..6170)
                     /label=AmpR promoter
     terminator      complement(6455..6502)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(6506..7868,1..65))
                     /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"
ORIGIN
        1 agcgcagcac tagaaaacac gcttttcgcc caatccgccg cacgatgcag tgcactggta
       61 atcatatgta tatctccttc ttaaagttaa caaaattatt tctagagggg aattgttatc
      121 cgctcacaat tcccctatag tgagtcgtat taatatcggc attttctttt gcgtttctgt
      181 ctcttgatca gatctggcca cctaggccga attcccgggg atccggtgat tgattgagca
      241 agctttatgc ttgtaaaccg ttttgtgaaa aaatttttaa aataaaaaag gggacctcta
      301 gggtccccaa ttaattagta atataatcta ttaaaggtca ttcaaaaggt catccaccgg
      361 atcaccttac caagccctcg ctagattgtt aatgcggatg ttgcgattac ttcgcccaac
      421 tattgcgata acaagaaaag cgcctttcat gatatatctc ccaattttgt gtagggctta
      481 ttatgcacgc ttaaaaataa taaaagcgac ttgacctgat agtttggctg tgagcaatta
      541 tgtgcttagt gcatctaacg cttgagttaa ccgcgccgcg aagcggcgtc ggcttgaacg
      601 aattgttaga cattatttgc cgactacctt ggtgattcgc ctttcacgta gtggacaaaa
      661 tcaaccaact gatctgcgcg agcttcacgc tgccgcaagc atcagggcgc aagggctgct
      721 aaaggaagcg gaacacgtag aaagccagtc cgcagaaacg gtgctacccc ggatgaatgt
      781 cagctactgg gctatctgga caagggaaaa cgcaagcgca aagagaaaga ggtagcttgc
      841 agtgggctta catgacgata gctagactgg gcggttttat ggacagcaag cgaaccggaa
      901 ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct
      961 ttcttgccgc caaggatctg atggcgcagg ggatcaagat ctgatcaaga gacaggatga
     1021 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     1081 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     1141 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     1201 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     1261 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     1321 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     1381 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     1441 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     1501 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     1561 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     1621 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     1681 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     1741 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     1801 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     1861 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     1921 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     1981 tggagttctt cgcccacccc gggctcgatc ccctcgcgag ttggttcagc tgctgcctga
     2041 ggctggacga cctcgcggag ttctaccggc agtgcaaatc cgtcggcatc caggaaacca
     2101 gcagcggcta tccgcgcatc catgcccccg aactgcagga gtggggaggc acgatggccg
     2161 ctttggtcga cccggacggg acggatcagt gagggtttgc aactgtgggt caaggatctg
     2221 gatttcgatc acggcacgat catcgtcggg agggcaaggg ctccaaggat cgggccttga
     2281 tgttaccgag agcttggtac ccagtctgtg tgagcagggg aattgatccg gtggatgacc
     2341 ttttgaatga cctttaatag attatattac taattaattg gggaccctag aggtcccctt
     2401 ttttatttta aaaatttttt cacaaaacgg tttacaagca taaagcttgc tcaatcaatc
     2461 accggatccc cgggaattcg tcgacaagct gcggccgcct aggccgtggc cgaacttgtg
     2521 tataagagtc aggagctctc ccgggaattc cacaaattgt tatccgctca caattccaca
     2581 tgtggaattc cacatgtgga attcccatgt cagccgttaa gtgttcctgt gtcactcaaa
     2641 attgctttga gaggctctaa gggcttctca gtgcgttaca tccctggctt gttgtccaca
     2701 accgttaaac cttaaaagct ttaaaagcct tatatattct tttttttctt ataaaactta
     2761 aaaccttaga ggctatttaa gttgctgatt tatattaatt ttattgttca aacatgagag
     2821 cttagtacgt gaaacatgag agcttagtac gttagccatg agagcttagt acgttagcca
     2881 tgagggttta gttcgttaaa catgagagct tagtacgtta aacatgagag cttagtacgt
     2941 gaaacatgag agcttagtac gtactatcaa caggttgaac tgctgatctt cagatcctct
     3001 acgccggacg catcgtggcc ggatccagcc gaccaggctt tccacgcccg cgtgccgctc
     3061 catgtcgttc gcgcggttct cggaaacgcg ctgccgcgtt tcgtgattgt cacgctcaag
     3121 cccgtagtcc cgttcgagcg tcgcgcagag gtcagcgagg gcgcggtagg cccgatacgg
     3181 ctcatggatg gtgtttcggg tcgggtgaat cttgttgatg gcgatatgga tgtgcaggtt
     3241 gtcggtgtcg tgatgcacgg cactgacgcg ctgatgctcg gcgaagccaa gcccagcgca
     3301 gatgcggtcc tcaatcgcgc gcaacgtctc cgcgtcgggc ttctctcccg cgcggaagct
     3361 aaccagcagg tgataggtct tgtcggcctc ggaacgggtg ttgccgtgct gggtcgccat
     3421 cacctcggcc atgacagcgg gcagggtgtt tgcctcgcag ttcgtgacgc gcacgtgacc
     3481 caggcgctcg gtcttgcctt gctcgtcggt gatgtacttc accagctccg cgaagtcgct
     3541 cttcttgatg gagcgcatgg ggacgtgctt ggcaatcacg cgcacccccc ggccgtttta
     3601 gcggctaaaa aagtcatggc tctgccctcg ggcggaccac gcccatcatg accttgccaa
     3661 gctcgtcctg cttctcttcg atcttcgcca gcagggcgag gatcgtggca tcaccgaacc
     3721 gcgccgtgcg cgggtcgtcg gtgagccaga gtttcagcag gccgcccagg cggcccaggt
     3781 cgccattgat gcgggccagc tcgcggacgt gctcatagtc cacgacgccc gtgattttgt
     3841 agccctggcc gacggccagc aggtaggccg acaggctcat gccggccgcc gccgcctttt
     3901 cctcaatcgc tcttcgttcg tctggaaggc agtacacctt gataggtggg ctgcccttcc
     3961 tggttggctt ggtttcatca gccatccgct tgccctcatc tgttacgccg gcggtagccg
     4021 gccagcctcg cagagcagga ttcccgttga gcaccgccag gtgcgaataa gggacagtga
     4081 agaaggaaca cccgctcgcg ggtgggccta cttcacctat cctgcccggc tgacgccgtt
     4141 ggatacacca aggaaagtct acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa
     4201 aggatggata taccgaaaaa atcgctataa tgaccccgaa gcagggttat gcagcggaaa
     4261 agcgctgctt ccctgctgtt ttgtggaata tctaccgact ggaaacaggc aaatgcagga
     4321 aattactgaa ctgaggggac aggcgagaga cgatgccaaa gagctacacc gacgagctgg
     4381 ccgagtgggt tgaatcccgc gcggccaaga agcgccggcg tgatgaggct gcggttgcgt
     4441 tcctggcggt gagggcggat gtcgaggcgg cgttagcgtc cggctatgcg ctcgtcacca
     4501 tttgggagca catgcgggaa acggggaagg tcaagttctc ctacgagacg ttccgctcgc
     4561 acgccaggcg gcacatcaag gccaagcccg ccgatgtgcc cgcaccgcag gccaaggctg
     4621 cggaacccgc gccggcaccc aagacgccgg agccacggcg gccgaagcag gggggcaagg
     4681 ctgaaaagcc ggcccccgct gcggccccga ccggcttcac cttcaaccca acaccggaca
     4741 aaaaggatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
     4801 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
     4861 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc
     4921 gccatctcct tgctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     4981 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     5041 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     5101 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
     5161 cataaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5221 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5281 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5341 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5401 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5461 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5521 gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5581 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5641 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5701 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5761 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5821 acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5881 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5941 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6001 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6061 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6121 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6181 cgaaaagtgc cacctgcagt tcacttacac cgcttctcaa cccggtacgc accagaaaat
     6241 cattgatatg gccatgaatg gcgttggatg ccgggcaaca gcccgcatta tgggcgttgg
     6301 cctcaacacg attttacgtc acttaaaaaa ctcaggccgc agtcggtaac ctcgcgcata
     6361 cagccgggca gtgacgtcat cgtctgcgcg gaaatggacg aacagtgggg ctatgtcggg
     6421 gctaaatcgc gccagcgctg gctgttttac gcgtcaaaaa acccctcaag acccgtttag
     6481 aggccccaag gggttatgct agttagattt taatgccctg cgccatcagg tctttcgcgg
     6541 ccagaaagcc atccagtttg ctttgcagcg cttcccaacc ttcccacagc gcaccccagc
     6601 tcgcaatgcc ggtacgtttg ctatccataa agccgcccag acgcgcaatc gccatatacg
     6661 cccattgcag gctgcccgct ttttctttgc gtttgcgttt gcctttatcc agatagccca
     6721 gcagttggca ttcatccggg gtcagcacgg tttccgcgct ctggctttca acgtgttccg
     6781 cttctttcag caggccctgc gcacgcagtg cttgcggcgg agtaaaagat tcacgcagtt
     6841 gcagcagacg caccgccaca aagctcagaa tgctcaccat acgttccagg ttatccggtt
     6901 cttccatacg ctgacgttcc gcacccgcac ccgttttcca cgctttgtga aattcttcaa
     6961 tgcgccaacg atgggtataa atatcaatca cacgcagcgc ttgggccaga ctttccaccg
     7021 gctcgctggt cagcagcagc catttcagcg gggtttcgcc tttcggcgga ttaatttctt
     7081 cggccagcac cgcgttcagg gtaatgttgc cctgtttcag ggtaatacgg ccgctacgca
     7141 ggctcaggct cgctttacgc gccggacggt ttttacgttt gccacgttta tccaccacgc
     7201 ctttctgcgg aatgctaatc tgatagccgc ccagttccgg ctggtttttc aggtgatcat
     7261 acagatacag gccgctttcc acatctttac gcggatgttt gctacgcacc acaaaacgtt
     7321 cgttatgggc cagtttatct tgcagatacg catgaatatc cgcttcacga tcgcacaccg
     7381 caatcacgtt gctcatcatg ctgcccattc tcagacgcga agttgcagca gcggccagcc
     7441 atttgccgct ttctttttca tccgcatccg ccggatcatc cggacgcatc caccattctt
     7501 gatgcagcag gcccacggta cgaaaggtgg tcgcttccag cagcagcacg ctatgcaccc
     7561 accaaccacg gcttttatcc tgaatgctac ccagtttgcc cagttcttcc gccacctgat
     7621 gacgatagct cagagaggtg gtatcttcaa ttgccagcag ttccggaaat tcctgggcca
     7681 gtttcacggt ctgcatggca cccgctttac gaatcgcttc cgcgctcacg ttcggattac
     7741 gaataaaacg atacgcgcct tcctgcatgg ctttgctgcc ttcgctgcta atggtaatgc
     7801 ttttgccgct atatttggcc agttgcgccg caacattcac cagacgcgcg gtacgacgcg
     7861 gatcaccc
//
