LOCUS       V046779                 7621 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046779
VERSION     V046779
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..2280
                     /label="Ppp3cb(NM_008914)"
                     /note="Ppp3cb(NM_008914)"
                     /gene="Ppp3cb"
     CDS             2287..2313
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2317..2343
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2347..2373
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2397..2985
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3051..3099
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3301..3729
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3743..4072
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4139..4933
                     /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    5109..5230
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5279..5295)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5303..5319
                     /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(5327..5357)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5372..5393
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5681..6269)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6440..7300)
                     /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(7301..7405)
                     /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 ccaccatggc cgccccggag
      721 ccggcccggg ccgcgccgcc cccgcccccg cccccaccgc cccctctcgg ggccgaccgc
      781 gtcgtcaaag ctgttccttt tcccccaact catcggctga catctgaaga agtgtttgat
      841 atggatggga tacccagggt tgatgttctg aagaaccact tggtaaaaga agggcgggtg
      901 gatgaagaaa ttgcactaag aattatcaat gagggtgctg ccatacttcg gcgggagaaa
      961 accatgatag aagtagaagc tccaattaca gtgtgtggtg acatccatgg ccaatttttt
     1021 gatctgatga aactttttga agtaggagga tcacctgcta atacacgata cctttttctt
     1081 ggtgattatg tggacagagg ttattttagt atagagtgtg tcttatattt atgggtcttg
     1141 aagattctat acccaagcac attattcctt ctgagaggca accatgaatg cagacacctt
     1201 actgaatatt ttacctttaa gcaggaatgt aaaattaaat attcagaaag agtctatgaa
     1261 gcttgtatgg aggcttttga cagcttgccc cttgctgcac ttctaaacca acaatttctt
     1321 tgtgttcatg gtggactttc accagaaata cacacactgg atgatattag gagattagat
     1381 agatttaaag agccacctgc atttggacca atgtgtgact tgctatggtc tgatccttct
     1441 gaagactttg gaaatgaaaa atcacaagaa cattttagtc ataatacagt tcgaggatgt
     1501 tcttattttt ataactatcc agcagtgtgt gaatttttgc aaaacaataa tttgttatcg
     1561 attattagag ctcatgaagc tcaagatgca ggctatagaa tgtacagaaa aagtcaaact
     1621 acagggtttc cttcattaat aacaattttt tcggcaccta attacttaga tgtctacaat
     1681 aataaagctg ctgtactaaa gtatgaaaat aatgtgatga acattcgaca gtttaattgc
     1741 tctccacatc cttattggtt gcccaatttt atggatgtct ttacatggtc tttaccattt
     1801 gttggagaaa aagtgacaga aatgttggta aatgttctga gtatttgttc tgatgatgaa
     1861 ctaatgacag aaggtgaaga ccagtttgat gtaggttcag ctgcagcccg gaaagaaatc
     1921 ataagaaaca agatccgagc aattggcaag atggcaagag tcttctctgt tctcagggag
     1981 gagagtgaaa gcgtgctgac actcaagggc ctgactccca cagggatgtt gcctagtgga
     2041 gtgttggctg gaggacggca gaccttgcaa agtgccacag ttgaggctat tgaggctgaa
     2101 aaagcaatac gaggattctc tccaccacat agaatctgca gttttgaaga ggcaaagggt
     2161 ttggatagga tcaatgagag aatgccaccc cggaaagatg ctgtgcagca agatggtttc
     2221 aattccctga acaccgcaca taccactgag aaccacggga ctggcaacca tagtgcccag
     2281 ctcgagtacc catacgacgt acctgattat gcaacttatc cttatgatgt acctgattat
     2341 gctagctatc cttatgatgt cccagactac gcatagtgag ggcccgatat ctcgacaatc
     2401 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     2461 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     2521 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     2581 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     2641 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     2701 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     2761 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     2821 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     2881 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     2941 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag
     3001 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     3061 aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     3121 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     3181 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     3241 tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc
     3301 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3361 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3421 cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta
     3481 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3541 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3601 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3661 aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3721 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3781 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3841 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3901 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3961 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     4021 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     4081 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     4141 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4201 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4261 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4321 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4381 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4441 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4501 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4561 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4621 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4681 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4741 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4801 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4861 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4921 cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc
     4981 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5041 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5101 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5161 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5221 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5281 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5341 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5401 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5461 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5521 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5581 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5641 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5701 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5761 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5821 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     5881 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5941 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6001 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6061 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6121 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6181 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6241 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6301 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6361 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6421 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6481 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6541 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6601 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6661 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6721 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6781 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6841 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6901 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6961 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7021 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7081 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7141 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7201 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7261 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7321 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7381 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     7441 gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc
     7501 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     7561 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     7621 g
//
