LOCUS       V027412                 7675 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027412
VERSION     V027412
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..3114
                     /label="HGF(NM_000601)"
                     /note="HGF(NM_000601)"
                     /gene="HGF"
     CDS             3121..3147
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3151..3177
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3181..3207
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    3242..3466
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3512..3940
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3954..4283
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4350..5144
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5318..5439
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5488..5504)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5512..5528
                     /bound_moiety="lac repressor encoded by lacI"
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5536..5566)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5581..5602
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5890..6475)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6646..7506)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7507..7611)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atgtgggtga ccaaactcct gccagccctg
      961 ctgctgcagc atgtcctcct gcatctcctc ctgctcccca tcgccatccc ctatgcagag
     1021 ggacaaagga aaagaagaaa tacaattcat gaattcaaaa aatcagcaaa gactacccta
     1081 atcaaaatag atccagcact gaagataaaa accaaaaaag tgaatactgc agaccaatgt
     1141 gctaatagat gtactaggaa taaaggactt ccattcactt gcaaggcttt tgtttttgat
     1201 aaagcaagaa aacaatgcct ctggttcccc ttcaatagca tgtcaagtgg agtgaaaaaa
     1261 gaatttggcc atgaatttga cctctatgaa aacaaagact acattagaaa ctgcatcatt
     1321 ggtaaaggac gcagctacaa gggaacagta tctatcacta agagtggcat caaatgtcag
     1381 ccctggagtt ccatgatacc acacgaacac agctttttgc cttcgagcta tcggggtaaa
     1441 gacctacagg aaaactactg tcgaaatcct cgaggggaag aagggggacc ctggtgtttc
     1501 acaagcaatc cagaggtacg ctacgaagtc tgtgacattc ctcagtgttc agaagttgaa
     1561 tgcatgacct gcaatgggga gagttatcga ggtctcatgg atcatacaga atcaggcaag
     1621 atttgtcagc gctgggatca tcagacacca caccggcaca aattcttgcc tgaaagatat
     1681 cccgacaagg gctttgatga taattattgc cgcaatcccg atggccagcc gaggccatgg
     1741 tgctatactc ttgaccctca cacccgctgg gagtactgtg caattaaaac atgcgctgac
     1801 aatactatga atgacactga tgttcctttg gaaacaactg aatgcatcca aggtcaagga
     1861 gaaggctaca ggggcactgt caataccatt tggaatggaa ttccatgtca gcgttgggat
     1921 tctcagtatc ctcacgagca tgacatgact cctgaaaatt tcaagtgcaa ggacctacga
     1981 gaaaattact gccgaaatcc agatgggtct gaatcaccct ggtgttttac cactgatcca
     2041 aacatccgag ttggctactg ctcccaaatt ccaaactgtg atatgtcaca tggacaagat
     2101 tgttatcgtg ggaatggcaa aaattatatg ggcaacttat cccaaacaag atctggacta
     2161 acatgttcaa tgtgggacaa gaacatggaa gacttacatc gtcatatctt ctgggaacca
     2221 gatgcaagta agctgaatga gaattactgc cgaaatccag atgatgatgc tcatggaccc
     2281 tggtgctaca cgggaaatcc actcattcct tgggattatt gccctatttc tcgttgtgaa
     2341 ggtgatacca cacctacaat agtcaattta gaccatcccg taatatcttg tgccaaaacg
     2401 aaacaattgc gagttgtaaa tgggattcca acacgaacaa acataggatg gatggttagt
     2461 ttgagataca gaaataaaca tatctgcgga ggatcattga taaaggagag ttgggttctt
     2521 actgcacgac agtgtttccc ttctcgagac ttgaaagatt atgaagcttg gcttggaatt
     2581 catgatgtcc acggaagagg agatgagaaa tgcaaacagg ttctcaatgt ttcccagctg
     2641 gtatatggcc ctgaaggatc agatctggtt ttaatgaagc ttgccaggcc tgctgtcctg
     2701 gatgattttg ttagtacgat tgatttacct aattatggat gcacaattcc tgaaaagacc
     2761 agttgcagtg tttatggctg gggctacact ggattgatca actatgatgg cctattacga
     2821 gtggcacatc tctatataat gggaaatgag aaatgcagcc agcatcatcg agggaaggtg
     2881 actctgaatg agtctgaaat atgtgctggg gctgaaaaga ttggatcagg accatgtgag
     2941 ggggattatg gtggcccact tgtttgtgag caacataaaa tgagaatggt tcttggtgtc
     3001 attgttcctg gtcgtggatg tgccattcca aatcgtcctg gtatttttgt ccgagtagca
     3061 tattatgcaa aatggataca caaaattatt ttaacatata aggtaccaca gtcactcgag
     3121 tacccatacg acgtacctga ttatgcaact tatccttatg atgtacctga ttatgctagc
     3181 tatccttatg atgtcccaga ctacgcatga gggcccgttt aaacccgctg atcagcctcg
     3241 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3301 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3361 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3421 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3481 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3541 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3601 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3661 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3721 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3781 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3841 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3901 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     3961 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4021 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4081 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4141 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4201 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4261 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4321 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4381 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4441 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4501 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4561 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4621 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4681 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4741 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4801 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4861 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4921 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4981 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5041 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5101 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5161 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5221 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5281 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5341 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5401 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5461 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5521 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5581 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5641 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5701 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5761 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5821 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5881 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5941 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6001 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6061 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6121 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6181 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6241 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6301 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6361 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6421 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6481 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6541 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6601 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6661 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6721 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6781 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6841 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6901 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6961 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7021 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7081 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7141 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7201 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7261 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7321 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7381 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7441 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7501 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7561 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7621 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggt
//
