LOCUS       V046918                 9106 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046918
VERSION     V046918
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..4545
                     /label="Atp2b4(NM_213616)"
                     /note="Atp2b4(NM_213616)"
                     /gene="Atp2b4"
     CDS             4552..4578
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4582..4608
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4612..4638
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    4673..4897
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4943..5371
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5385..5714
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5781..6575
                     /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    6749..6870
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6919..6935)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6943..6959
                     /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(6967..6997)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7012..7033
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7321..7906)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8077..8937)
                     /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(8938..9042)
                     /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 atgacgaatc caccaggaca aagcgtgtca
      961 gccaacacag tggctgagag ccatgaaggg gagtttggct gcaccttaat ggacctgcga
     1021 aagctcatgg agcttcgtgg agctgatgca gtggcccaga tcagtgccca ctacggaggg
     1081 gtacaggaaa tctgcactag actgaaaacc tcccctatag aaggtctatc tgggaaccct
     1141 gcagatttgg agaagcgtag acttgttttt ggaaagaacg tgatacctcc aaaaaggccc
     1201 aagactttct tagaattagt gtgggaagcc ctgcaggatg tcacgctcat catcctagag
     1261 atcgcagcca tcatctccct ggtcctgtcc ttctaccgac ctccgggtgg agataatgaa
     1321 atctgtggtc acattgcaag tagcccagaa gaagaggagg aaggagaaac tggctggatt
     1381 gagggggcgg ccattctcgc ctcggtgatc attgtggtac tcgtgacagc cttcaatgac
     1441 tggagcaaag agaagcagtt ccgggggctg cagagtcgca tcgaactgga gcaaaaattc
     1501 tccattatcc ggaatggtca gcttatccaa ctcccagtgg ctgagattgt ggtgggagat
     1561 attgcccaga tcaaatacgg tgacctgctg cctgcagatg gaattctaat ccagggaaat
     1621 gatctgaaga ttgacgagag ctctctgaca ggagaatcag atcatgtcaa gaagactctg
     1681 gacaaagacc ccatgttgct ctcggggact cacgtgatgg aaggttctgg acggatggta
     1741 gtgactgctg tgggggtcaa ctcccagact ggaatcatct tcaccctctt aggagctagt
     1801 gaggaagaag atgatgatga caagaagaag aaaggtaaaa aacaaggagc ccctgaaaat
     1861 cgcaacaaag caaagaccca ggatggagtg gccttggaaa tccagccact caacagccaa
     1921 gaggggctcg acagtgagga caaggagaag aaaatagcca ggatacccaa gaaagagaag
     1981 tcggtgctgc aaggcaaact cacacgcctg gctgtgcaga tcgggaaggc gggtctgatc
     2041 atgtctgtcc tcacagttgt catcttgatt ctgtactttg tggttgacaa tttcgtgatc
     2101 cagcgccgag aatggcttcc tgagtgcact cctgtctata tccagtattt tgtcaagttc
     2161 ttcatcatcg gagtcactgt attggtagtg gctgtgccag aggggctgcc actggctgtc
     2221 accatctcac tagcctactc tgtaaagaaa atgatgaagg acaacaactt ggtacggcac
     2281 ttggatgctt gtgagacaat gggcaatgcc acagccattt gctccgataa gacaggcaca
     2341 ctgaccatga accgcatgac cgtggtgcaa gcttacatcg ggggcactca ctaccgccag
     2401 atcccacagc ccgacgtctt cccacccaag gttctggagc tcatcgtcaa tggcatctct
     2461 atcaactgtg cttacacgtc taagattcag cctccagaaa aggagggagg cctaccccga
     2521 caagtaggca acaagacaga gtgtgggctg ctgggctttg tcacagacct gaagcaggac
     2581 taccaagcag ttcgcaatga ggtgcctgag gagaagctct ttaaggtgta caccttcaac
     2641 tcagtgcgca agtccatgag cactgtcatc aggaagccgg aagggggctt ccgcatgttc
     2701 agcaagggcg cctctgagat aatgctgcgc aggtgtgatc gaatcctgaa caaggaagga
     2761 gaaatcaaat cgttcagaag taaggaccga gataatatgg tacgcaatgt cattgagccc
     2821 atggccagtg aagggctccg gactatctgc ttagcttacc gagattttga tggcacagag
     2881 ccctcgtggg acattgaggg tgaaatcctc acttcgctga tctgcattgc agtggtgggc
     2941 attgaggatc ctgtgcgccc ggaggtacca gatgcaattg ccaaatgcaa acgggctggc
     3001 attactgtga ggatggtgac aggtgataat gtcaacacag cccgggccat tgccaccaaa
     3061 tgtggcattc tgactcctaa agatgacttc ttgtgcctgg aaggcaaaga attcaacagt
     3121 cttatccgaa atgagaaggg tgaggttgaa caagaaaagc tggacaagat ctggcccaag
     3181 cttcgggtac tggcacgatc ttctcccaca gacaagcaca cattggtgaa aggtatcatt
     3241 gacagcactg ctggggaaca gcggcaggta gtggctgtca ccggtgatgg aacaaatgat
     3301 ggacctgctc tgaagaaagc agatgtcggg tttgctatgg gcatcgcagg cacagatgtg
     3361 gctaaggagg cgtcagacat catcctgacc gatgacaact tcaccagcat tgtgaaggca
     3421 gtgatgtggg gacggaatgt gtatgacagc atttccaagt tcctacagtt ccagctcact
     3481 gtcaatgtgg tggccgtgat tgtggctttc actggagcct gcatcactca ggattcccca
     3541 ctgaaagcgg tgcagatgtt gtgggttaac ctgattatgg acacgtttgc ttccctggcc
     3601 ctggccacag agccccccac cgagtctctg ttgaggcggc gcccctatgg gcgaaacaag
     3661 cctctgatct cccgcactat gatgaagaac atcttgggcc atgctgtgta tcagctccta
     3721 atcgtcttcc tcctcgtctt tgctggcgac acactgttcg acattgacag tgggaggaag
     3781 gcacctctca attcgccacc cagccagcac tacaccattg ttttcaacac gtttgtgctg
     3841 atgcaactct tcaacgaaat caacgctcgg aagatccacg gtgagaagaa cgtgtttgca
     3901 ggcgtctacc gcaacatcat tttctgcact gtagttttgg gcacattctt ctgccagatc
     3961 atgattgttg aactgggagg gaagcccttc agctgcacaa gcctgaccat ggagcagtgg
     4021 atgtggtgtc tctttatagg gattggagaa cttttgtggg gtcaggtcat ctctgcaata
     4081 cctaccaagt ctctgaagtt cctgaaggag gctggccacg gcagtgacaa agaggacatc
     4141 agcagggata ctgagggaat ggacgagatt gaccttgccg agatggagct tcgccgaggc
     4201 cagatcctct gggtccgtgg cctgaaccgg attcagactc agatcagagt cgtcaaactg
     4261 ttccataata accatgaagt tgctcataaa cccaagaatc gtagctccat ccacaccttc
     4321 atgacccaac ctgaataccc cgcagatgat gaattgtcac aatccttcct ggatatccaa
     4381 gagggcaatc ctgaattggt ttctaaggcg gggacgagtg tgctcctgtt ggatggtgaa
     4441 gctgcttcgc acgacaacat aaataacaac gccgtggact gccaccaagt gcaaatcgtt
     4501 gcctcccatt ctgacagccc tctgccaagc ctggagacac cggtcctcga gtacccatac
     4561 gacgtacctg attatgcaac ttatccttat gatgtacctg attatgctag ctatccttat
     4621 gatgtcccag actacgcatg agggcccgtt taaacccgct gatcagcctc gactgtgcct
     4681 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     4741 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     4801 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     4861 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     4921 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4981 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     5041 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     5101 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     5161 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     5221 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     5281 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     5341 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     5401 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5461 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5521 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     5581 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5641 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5701 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     5761 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     5821 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5881 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5941 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     6001 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     6061 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     6121 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     6181 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     6241 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     6301 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     6361 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     6421 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     6481 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     6541 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     6601 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     6661 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     6721 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     6781 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6841 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     6901 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6961 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     7021 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     7081 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     7141 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     7201 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     7261 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7321 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7381 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7441 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7501 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7561 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7621 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7681 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7741 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     7801 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt
     7861 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7921 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7981 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     8041 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     8101 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     8161 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     8221 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     8281 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8341 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     8401 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     8461 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     8521 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8581 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8641 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     8701 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8761 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8821 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8881 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8941 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     9001 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     9061 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggt
//
