LOCUS       V019147                 9020 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019147
VERSION     V019147
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             683..3676
                     /label="Atp2a2(NM_009722)"
                     /note="Atp2a2(NM_009722)"
                     /gene="Atp2a2"
     CDS             3683..3772
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3796..4384
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4450..4498
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4700..5128
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5142..5471
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5538..6332
                     /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    6508..6629
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6678..6694)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6702..6718
                     /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(6726..6756)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6771..6792
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7080..7668)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7839..8699)
                     /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(8700..8804)
                     /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 ggatccgcca ccatggagaa cgctcacaca aagaccgtgg aggaggtgct
      721 gggccacttc ggggtcaacg agagcacggg gctgagcttg gagcaggtca agaagctcaa
      781 ggagagatgg ggctccaacg aattgccggc tgaagaagga aaaaccttgc tggaacttgt
      841 gattgagcag tttgaagact tactagttag aattttactg ctggcagcat gtatatcttt
      901 cgttttggct tggttcgagg aaggggaaga aacgattaca gcctttgtag agccgtttgt
      961 aattctgctt atcttggtag ccaatgcaat cgtgggtgtg tggcaggaaa gaaatgctga
     1021 aaatgcaata gaagctctta aggaatatga gcctgaaatg ggcaaagtgt atcgacagga
     1081 cagaaagagt gtgcaacgaa ttaaagctaa agacatagtt cctggtgata tagtggaaat
     1141 tgctgttggt gacaaagttc ctgctgatat tagattgaca tccatcaagt ctacaactct
     1201 aagagtcgac cagtcaattc ttacaggtga atctgtctcc gtcatcaagc atactgaccc
     1261 tgtccctgac ccccgagctg ttaatcaaga caaaaagaac atgctctttt ctggtacaaa
     1321 cattgctgct gggaaagcta tgggagtggt ggtggcaact ggagttaata ctgagatcgg
     1381 caagatccgg gatgaaatgg tggcaacaga acaggagaga acacccctac agcagaagct
     1441 agacgagttt ggggagcagc tttccaaagt tatctccctc atttgcattg cagtctggat
     1501 catcaacatt gggcatttca atgacccagt tcatggtggc tcctggatca ggggtgccat
     1561 ctactacttt aagattgccg tggccctggc tgttgccgca atccctgagg gtctgcctgc
     1621 tgtcatcacc acctgcttag ctcttggaac tcgtaggatg gcaaagaaaa atgctatcgt
     1681 tcgaagtctg ccttctgtgg agacccttgg ttgtacttct gttatctgct cagataagac
     1741 aggcacactt accacaaacc agatgtccgt gtgcaggatg ttcattctgg acaaagtaga
     1801 aggtgacact tgttccctta atgagttcag cataactgga tccacatatg caccaattgg
     1861 agaagtgcaa aaggatgata agccagtgaa gtgccatcag tatgacgggc ttgtagagtt
     1921 agccaccatc tgtgctctgt gtaatgactc tgctttggat tataatgagg caaagggtgt
     1981 gtatgagaaa gttggagaag ctaccgagac tgctctcacg tgcctggtgg agaagatgaa
     2041 tgtatttgat actgagctga aggggctttc taaaatagag cgtgcaaacg cctgcaactc
     2101 ggtcataaag cagctgatga agaaggagtt cactctggag ttttcacggg atagaaaatc
     2161 aatgtccgtc tattgtaccc caaacaagcc aagccggaca tccatgagca agatgtttgt
     2221 gaagggggct ccagaaggtg tcatcgatag gtgcacccac atccgagttg gaagtaccaa
     2281 ggtccccatg actcctggtg tcaaacagaa gattatgtct gtcattcggg agtggggcag
     2341 tggcagcgac acgctacggt gcctggctct ggccactcat gacaacccac tgaagagaga
     2401 ggagatgcac ctggaagact ctgctaactt catcaaatac gagaccaacc tgactttcgt
     2461 cggctgtgtg ggcatgctgg atcctcccag gattgaagta gcctcttctg tgaagctgtg
     2521 ccggcaagca ggcatccggg tcatcatgat cactggagac aacaaaggca ccgctgtggc
     2581 catctgtcgc cgcattggca tctttgggca ggatgaggat gtgacatcaa aggcttttac
     2641 agggcgagag tttgatgaat taagcccttc agcccagaga gatgcctgct taaatgcccg
     2701 ctgttttgct cgagttgaac cttcccacaa gtctaagatt gttgagttcc ttcagtcctt
     2761 tgatgagatc acagctatga ctggtgatgg tgtgaatgat gctcctgctc tgaagaaatc
     2821 tgaaatcggg attgccatgg gctcagggac tgcagtggct aagactgctt ctgagatggt
     2881 cctggcagat gacaacttct ccaccattgt ggctgctgtt gaggaggggc gagccatcta
     2941 caacaacatg aagcagttca tccgctacct catctcatcc aacgtggggg aagtggtctg
     3001 tatcttcctg acggcagccc ttgggtttcc tgaggcttta attcctgtcc agttactctg
     3061 ggtcaatctg gtgacagatg gtctgcctgc cactgcgctg gggttcaatc ctccagacct
     3121 ggacatcatg aacaaacccc cccggaaccc aaaagaacca ctgatcagcg ggtggctctt
     3181 tttccgttac ctggctattg gctgttatgt tggcgctgcc accgtgggtg ctgctgcatg
     3241 gtggttcatc gctgctgacg gcggtccaag agtctccttc taccagctga gtcatttcct
     3301 acagtgtaag gaggacaacc cagacttcga tggagtggat tgtgcaatct ttgagtcccc
     3361 atatccaatg acaatggcac tttctgttct agtaaccata gagatgtgta atgccctcaa
     3421 cagcttgtct gaaaaccagt ctttgctgag gatgcccccc tgggagaata tctggctcgt
     3481 gggctccatc tgcttgtcca tgtcacttca cttcttgatc ctctacgtgg aacctttgcc
     3541 gctcattttc cagatcacac cgctgaatct gacccagtgg ctgatggtgc tgaaaatctc
     3601 cttgcctgtg atcctcatgg atgagacgct caagtttgtg gcccgaaact acctggaaca
     3661 acccgcaata ctggagctcg aggaacaaaa actaatatca gaggaagatt tggaacagaa
     3721 actaatcagt gaagaagatt tagagcaaaa gttaatttcc gaggaggact tatagtgagg
     3781 gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     3841 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     3901 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     3961 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     4021 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     4081 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     4141 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     4201 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     4261 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     4321 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     4381 ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4441 gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat
     4501 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     4561 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     4621 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg
     4681 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4741 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4801 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc
     4861 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4921 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4981 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     5041 gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag
     5101 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     5161 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5221 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5281 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     5341 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5401 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5461 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     5521 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     5581 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     5641 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     5701 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     5761 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     5821 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     5881 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5941 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     6001 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     6061 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     6121 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     6181 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     6241 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     6301 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc
     6361 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     6421 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     6481 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     6541 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6601 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     6661 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     6721 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6781 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6841 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6901 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6961 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     7021 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     7081 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     7141 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     7201 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     7261 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     7321 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7381 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7441 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7501 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7561 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7621 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7681 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7741 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7801 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7861 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7921 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7981 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     8041 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     8101 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     8161 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     8221 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     8281 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     8341 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8401 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     8461 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8521 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8581 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8641 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8701 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8761 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8821 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta
     8881 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     8941 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     9001 gcatgaagaa tctgcttagg
//
