LOCUS       V031896                 8838 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031896
VERSION     V031896
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..3518
                     /label="NFATC2(NM_012340)"
                     /note="NFATC2(NM_012340)"
                     /gene="NFATC2"
     CDS             3525..3590
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3614..4202
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4268..4316
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4518..4946
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4960..5289
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5356..6150
                     /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    6326..6447
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6496..6512)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6520..6536
                     /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(6544..6574)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6589..6610
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6898..7486)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7657..8517)
                     /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(8518..8622)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgaa cgcccccgag cggcagcccc
      781 aacccgacgg cggggacgcc ccaggccacg agcctggggg cagcccccaa gacgagcttg
      841 acttctccat cctcttcgac tatgagtatt tgaatccgaa cgaagaagag ccgaatgcac
      901 ataaggtcgc cagcccaccc tccggacccg cataccccga tgatgtcctg gactatggcc
      961 tcaagccata cagccccctt gctagtctct ctggcgagcc ccccggccga ttcggagagc
     1021 cggatagggt agggccgcag aagtttctga gcgcggccaa gccagcaggg gcctcgggcc
     1081 tgagccctcg gatcgagatc actccgtccc acgaactgat ccaggcagtg gggcccctcc
     1141 gcatgagaga cgcgggcctc ctggtggagc agccgcccct ggccggggtg gccgccagcc
     1201 cgaggttcac cctgcccgtg cccggcttcg agggctaccg cgagccgctt tgcttgagcc
     1261 ccgctagcag cggctcctct gccagcttca tttctgacac cttctccccc tacacctcgc
     1321 cctgcgtctc gcccaataac ggcgggcccg acgacctgtg tccgcagttt caaaacatcc
     1381 ctgctcatta ttcccccaga acctcgccaa taatgtcacc tcgaaccagc ctcgccgagg
     1441 acagctgcct gggccgccac tcgcccgtgc cccgtccggc ctcccgctcc tcatcgcctg
     1501 gtgccaagcg gaggcattcg tgcgccgagg ccttggttgc cctgccgccc ggagcctcac
     1561 cccagcgctc ccggagcccc tcgccgcagc cctcatctca cgtggcaccc caggaccacg
     1621 gctccccggc tgggtacccc cctgtggctg gctctgccgt gatcatggat gccctgaaca
     1681 gcctcgccac ggactcgcct tgtgggatcc cccccaagat gtggaagacc agccctgacc
     1741 cctcgccggt gtctgccgcc ccatccaagg ccggcctgcc tcgccacatc tacccggccg
     1801 tggagttcct ggggccctgc gagcagggcg agaggagaaa ctcggctcca gaatccatcc
     1861 tgctggttcc gcccacttgg cccaagccgc tggtgcctgc cattcccatc tgcagcatcc
     1921 cagtgactgc atccctccct ccacttgagt ggccgctgtc cagtcagtca ggctcttacg
     1981 agctgcggat cgaggtgcag cccaagccac atcaccgggc ccactatgag acagaaggca
     2041 gccgaggggc tgtcaaagct ccaactggag gccaccctgt ggttcagctc catggctaca
     2101 tggaaaacaa gcctctggga cttcagatct tcattgggac agctgatgag cggatcctta
     2161 agccgcacgc cttctaccag gtgcaccgaa tcacggggaa aactgtcacc accaccagct
     2221 atgagaagat agtgggcaac accaaagtcc tggagatacc cttggagccc aaaaacaaca
     2281 tgagggcaac catcgactgt gcggggatct tgaagcttag aaacgccgac attgagctgc
     2341 ggaaaggcga gacggacatt ggaagaaaga acacgcgggt gagactggtt ttccgagttc
     2401 acatcccaga gtccagtggc agaatcgtct ctttacagac tgcatctaac cccatcgagt
     2461 gctcccagcg atctgctcac gagctgccca tggttgaaag acaagacaca gacagctgcc
     2521 tggtctatgg cggccagcaa atgatcctca cggggcagaa ctttacatcc gagtccaaag
     2581 ttgtgtttac tgagaagacc acagatggac agcaaatttg ggagatggaa gccacggtgg
     2641 ataaggacaa gagccagccc aacatgcttt ttgttgagat ccctgaatat cggaacaagc
     2701 atatccgcac acctgtaaaa gtgaacttct acgtcatcaa tgggaagaga aaacgaagtc
     2761 agcctcagca ctttacctac cacccagtcc cagccatcaa gacggagccc acggatgaat
     2821 atgaccccac tctgatctgc agccccaccc atggaggcct ggggagccag ccttactacc
     2881 cccagcaccc gatggtggcc gagtccccct cctgcctcgt ggccaccatg gctccctgcc
     2941 agcagttccg cacggggctc tcatcccctg acgcccgcta ccagcaacag aacccagcgg
     3001 ccgtactcta ccagcggagc aagagcctga gccccagcct gctgggctat cagcagccgg
     3061 ccctcatggc cgccccgctg tcccttgcgg acgctcaccg ctctgtgctg gtgcacgccg
     3121 gctcccaggg ccagagctca gccctgctcc acccctctcc gaccaaccag caggcctcgc
     3181 ctgtgatcca ctactcaccc accaaccagc agctgcgctg cggaagccac caggagttcc
     3241 agcacatcat gtactgcgag aatttcgcac caggcaccac cagacctggc ccgcccccgg
     3301 tcagtcaagg tcagaggctg agcccgggtt cctaccccac agtcattcag cagcagaatg
     3361 ccacgagcca aagagccgcc aaaaacggac ccccggtcag tgaccaaaag gaagtattac
     3421 ctgcgggggt gaccattaaa caggagcaga acttggacca gacctacttg gatgatgagc
     3481 tgatagacac acaccttagc tggatacaaa acatattagg atccgactac aaagaccatg
     3541 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc
     3601 ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3661 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3721 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3781 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3841 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     3901 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3961 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     4021 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     4081 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     4141 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     4201 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     4261 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     4321 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     4381 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     4441 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     4501 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4561 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4621 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     4681 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4741 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4801 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4861 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     4921 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4981 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5041 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5101 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5161 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5221 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     5281 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     5341 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5401 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5461 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5521 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5581 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5641 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5701 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5761 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5821 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5881 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5941 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     6001 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     6061 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     6121 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     6181 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     6241 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     6301 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     6361 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6421 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     6481 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6541 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6601 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6661 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6721 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6781 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6841 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6901 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6961 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7021 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7081 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7141 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7201 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7261 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7321 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7381 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7441 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7501 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7561 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7621 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7681 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7741 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7801 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7861 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7921 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7981 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     8041 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     8101 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8161 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8221 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     8281 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8341 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8401 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8461 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8521 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8581 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8641 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
     8701 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     8761 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     8821 atgaagaatc tgcttagg
//
