LOCUS       V022770                 7270 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022770
VERSION     V022770
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             706..1929
                     /label="BMP4(NM_130850)"
                     /note="BMP4(NM_130850)"
                     /gene="BMP4"
     CDS             1936..1962
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1966..1992
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1996..2022
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2046..2634
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2700..2748
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2950..3378
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3392..3721
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3788..4582
                     /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    4758..4879
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4928..4944)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4952..4968
                     /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(4976..5006)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5021..5042
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5330..5918)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6089..6949)
                     /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(6950..7054)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgat tcctggtaac
      721 cgaatgctga tggtcgtttt attatgccaa gtcctgctag gaggcgcgag ccatgctagt
      781 ttgatacctg agacggggaa gaaaaaagtc gccgagattc agggccacgc gggaggacgc
      841 cgctcagggc agagccatga gctcctgcgg gacttcgagg cgacacttct gcagatgttt
      901 gggctgcgcc gccgcccgca gcctagcaag agtgccgtca ttccggacta catgcgggat
      961 ctttaccggc ttcagtctgg ggaggaggag gaagagcaga tccacagcac tggtcttgag
     1021 tatcctgagc gcccggccag ccgggccaac accgtgagga gcttccacca cgaagaacat
     1081 ctggagaaca tcccagggac cagtgaaaac tctgcttttc gtttcctctt taacctcagc
     1141 agcatccctg agaacgaggt gatctcctct gcagagcttc ggctcttccg ggagcaggtg
     1201 gaccagggcc ctgattggga aaggggcttc caccgtataa acatttatga ggttatgaag
     1261 cccccagcag aagtggtgcc tgggcacctc atcacacgac tactggacac gagactggtc
     1321 caccacaatg tgacacggtg ggaaactttt gatgtgagcc ctgcggtcct tcgctggacc
     1381 cgggagaagc agccaaacta tgggctagcc attgaggtga ctcacctcca tcagactcgg
     1441 acccaccagg gccagcatgt caggattagc cgatcgttac ctcaagggag tgggaattgg
     1501 gcccagctcc ggcccctcct ggtcaccttt ggccatgatg gccggggcca tgccttgacc
     1561 cgacgccgga gggccaagcg tagccctaag catcactcac agcgggccag gaagaagaat
     1621 aagaactgcc ggcgccactc gctctatgtg gacttcagcg atgtgggctg gaatgactgg
     1681 attgtggccc caccaggcta ccaggccttc tactgccatg gggactgccc ctttccactg
     1741 gctgaccacc tcaactcaac caaccatgcc attgtgcaga ccctggtcaa ttctgtcaat
     1801 tccagtatcc ccaaagcctg ttgtgtgccc actgaactga gtgccatctc catgctgtac
     1861 ctggatgagt atgataaggt ggtactgaaa aattatcagg agatggtagt agagggatgt
     1921 gggtgccgcc tcgagtaccc atacgacgta cctgattatg caacttatcc ttatgatgta
     1981 cctgattatg ctagctatcc ttatgatgtc ccagactacg catagtgagg gcccgatatc
     2041 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2101 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2161 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2221 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     2281 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     2341 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     2401 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     2461 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     2521 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     2581 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     2641 acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     2701 ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg
     2761 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     2821 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     2881 cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg
     2941 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3001 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3061 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt
     3121 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3181 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3241 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3301 ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac
     3361 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3421 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3481 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3541 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3601 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3661 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3721 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     3781 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3841 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3901 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3961 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4021 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4081 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4141 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4201 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4261 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     4321 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4381 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4441 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4501 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4561 ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga
     4621 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4681 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4741 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4801 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4861 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4921 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4981 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5041 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5101 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5161 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5221 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5281 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5341 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5401 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5461 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5521 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5581 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5641 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5701 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5761 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5821 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5881 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5941 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6001 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6061 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6121 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6181 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6241 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6301 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6361 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6421 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6481 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6541 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6601 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6661 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6721 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6781 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6841 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6901 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6961 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7021 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7081 acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc
     7141 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     7201 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     7261 tctgcttagg
//
