LOCUS       V022645                 8392 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022645
VERSION     V022645
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      901..910
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             910..933
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             943..3945
                     /label="ATP2A1(NM_173201)"
                     /note="ATP2A1(NM_173201)"
                     /gene="ATP2A1"
     polyA_signal    3992..4216
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4262..4690
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4704..5033
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5100..5894
                     /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    6068..6189
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6238..6254)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6262..6278
                     /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(6286..6316)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6331..6352
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6640..7225)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7396..8256)
                     /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(8257..8361)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gccaccatgg attacaagga tgacgacgat aagactaagc ttatggaggc cgctcatgct
      961 aaaaccacgg aggaatgttt ggcctatttt ggggtgagtg agaccacggg cctcaccccg
     1021 gaccaagtta agcggaatct ggagaaatac ggcctcaatg agctccctgc tgaggaaggg
     1081 aagaccctgt gggagctggt gatagagcag tttgaagacc tcctggtgcg gattctcctc
     1141 ctggccgcat gcatttcctt cgtgctggcc tggtttgagg aaggtgaaga gaccatcact
     1201 gcctttgttg aaccctttgt catcctcttg atcctcattg ccaatgccat cgtgggggtt
     1261 tggcaggagc ggaacgcaga gaacgccatc gaggccctga aggagtatga gccagagatg
     1321 gggaaggtct accgggctga ccgcaagtca gtgcaaagga tcaaggctcg ggacatcgtc
     1381 cctggggaca tcgtggaggt ggctgtgggg gacaaagtcc ctgcagacat ccgaatcctc
     1441 gccatcaaat ccaccacgct gcgggttgac cagtccatcc tgacaggcga gtctgtatct
     1501 gtcatcaaac acacggagcc cgttcctgac ccccgagctg tcaaccagga caagaagaac
     1561 atgcttttct cgggcaccaa cattgcagcc ggcaaggcct tgggcatcgt ggccaccact
     1621 ggtgtgggca ccgagattgg gaagatccga gaccaaatgg ctgccacaga acaggacaag
     1681 acccccttgc agcagaagct ggatgagttt ggggagcagc tctccaaggt catctccctc
     1741 atctgtgtgg ctgtctggct tatcaacatt ggccacttca acgaccccgt ccatgggggc
     1801 tcctggttcc gcggggccat ctactacttt aagattgccg tggccttggc tgtggctgcc
     1861 atccccgaag gtcttcctgc agtcatcacc acctgcctgg ccctgggtac ccgtcggatg
     1921 gcaaagaaga atgccattgt aagaagcttg ccctccgtag agaccctggg ctgcacctct
     1981 gtcatctgtt ccgacaagac aggcaccctc accaccaacc agatgtctgt ctgcaagatg
     2041 tttatcattg acaaggtgga tggggacatc tgcctcctga atgagttctc catcaccggc
     2101 tccacttacg ctccagaggg agaggtcttg aagaatgata agccagtccg gccagggcag
     2161 tatgacgggc tggtggagct ggccaccatc tgtgccctct gcaatgactc ctccttggac
     2221 ttcaacgagg ccaaaggtgt ctatgagaag gtcggcgagg ccaccgagac agcactcacc
     2281 accctggtgg agaagatgaa tgtgttcaac acggatgtga gaagcctctc gaaggtggag
     2341 agagccaacg cctgcaactc ggtgatccgc cagctaatga agaaggaatt caccctggag
     2401 ttctcccgag acagaaagtc catgtctgtc tattgctccc cagccaaatc ttcccgggct
     2461 gctgtgggca acaagatgtt tgtcaagggt gcccctgagg gcgtcatcga ccgctgtaac
     2521 tatgtgcgag ttggcaccac ccgggtgcca ctgacggggc cggtgaagga aaagatcatg
     2581 gcggtgatca aggagtgggg cactggccgg gacaccctgc gctgcttggc cctggccacc
     2641 cgggacaccc ccccgaagcg agaggaaatg gtcctggatg actctgccag gttcctggag
     2701 tatgagacgg acctgacatt cgtgggtgta gtgggcatgc tggaccctcc gcgcaaggag
     2761 gtcacgggct ccatccagct gtgccgtgac gccgggatcc gggtgatcat gatcactggg
     2821 gacaacaagg gcacagccat tgccatctgc cggcgaattg gcatctttgg ggagaacgag
     2881 gaggtggccg atcgcgccta cacgggccga gagttcgacg acctgcccct ggctgaacag
     2941 cgggaagcct gccgacgtgc ctgctgcttc gcccgtgtgg agccctcgca caagtccaag
     3001 attgtggagt acctgcagtc ctacgatgag atcacagcca tgacaggtga tggcgtcaat
     3061 gacgcccctg ccctgaagaa ggctgagatt ggcattgcca tgggatctgg cactgccgtg
     3121 gccaagactg cctctgagat ggtgctggct gacgacaact tctccaccat cgtagctgct
     3181 gtggaggagg gccgcgccat ctacaacaac atgaagcagt tcatccgcta cctcatttcc
     3241 tccaacgtgg gcgaggtggt ctgtatcttc ctgaccgctg ccctggggct gcctgaggcc
     3301 ctgatcccgg tgcagctgct atgggtgaac ttggtgaccg acgggctccc agccacagcc
     3361 ctgggcttca acccaccaga cctggacatc atggaccgcc ccccccggag ccccaaggag
     3421 cccctcatca gtggctggct cttcttccgc tacatggcaa tcgggggcta tgtgggtgca
     3481 gccaccgtgg gagcagctgc ctggtggttc ctgtacgctg aggatgggcc tcatgtcaac
     3541 tacagccagc tgactcactt catgcagtgc accgaggaca acacccactt tgagggcata
     3601 gactgtgagg tcttcgaggc ccccgagccc atgaccatgg ccctgtccgt gctggtgacc
     3661 atcgagatgt gcaatgcact gaacagcctg tccgagaacc agtccctgct gcggatgcca
     3721 ccctgggtga acatctggct gctgggctcc atctgcctct ccatgtccct gcacttcctc
     3781 atcctctatg ttgaccccct gccgatgatc ttcaagctcc gggccctgga cctcacccag
     3841 tggctcatgg tcctcaagat ctcactgcca gtcattgggc tcgacgaaat cctcaagttc
     3901 gttgctcgga actacctaga ggatccagaa gatgaaagaa ggaagtgact cgagtctaga
     3961 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     4021 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     4081 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     4141 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     4201 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     4261 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4321 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4381 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4441 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4501 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4561 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4621 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4681 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4741 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4801 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4861 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4921 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4981 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     5041 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     5101 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5161 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     5221 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     5281 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5341 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5401 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5461 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5521 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5581 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     5641 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5701 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5761 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5821 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5881 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5941 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     6001 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     6061 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     6121 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     6181 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     6241 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6301 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6361 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6421 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6481 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6541 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6601 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6661 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6721 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6781 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6841 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6901 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6961 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7021 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7081 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7141 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     7201 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7261 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7321 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7381 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7441 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7501 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7561 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7621 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7681 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7741 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7801 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7861 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7921 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7981 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8041 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8101 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8161 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8221 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8281 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8341 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
