LOCUS       V041036                 6624 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041036
VERSION     V041036
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..1304
                     /label="Card19(NM_026738)"
                     /note="Card19(NM_026738)"
                     /gene="Card19"
     CDS             1311..1376
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1400..1988
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2054..2102
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2304..2732
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2746..3075
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3142..3936
                     /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    4112..4233
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4282..4298)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4306..4322
                     /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(4330..4360)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4375..4396
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4684..5272)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5443..6303)
                     /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(6304..6408)
                     /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 ccaccatgac agatcagacc tactgtgacc
      781 ggctggtaca ggacactcct ttcctgacgg gacagggtcg cctgagcgag caacaagtgg
      841 acaggatcat cctccagctg aaccgttact acccacagat ccttaccaac aaggaggctg
      901 aaaagttccg gaaccccaag gcatccctgc gcgtacggct ctgtgacctc ctgagccatc
      961 tacagcagcg tggggaacgg cactgtcagg aattttaccg agctctgtac atccatgccc
     1021 agcccctgca cagccacctg cccagccgct actctcctca aaactcagat tgcagagagc
     1081 tggactgggg catcgagagc cgtgagctga gtgacagggg acccatgagc ttcctggcag
     1141 gcctgggcct ggcagcgggc ctggccctcc tcctctactg ctgccctcca gaccccaaag
     1201 tgctgcccgg aacccggcgc gtccttgcct tctcacctgt catcattgac cggcatgtca
     1261 gccgctacct gctggctttc ctggcagatg acctcggagg actcggatcc gactacaaag
     1321 accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt
     1381 gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     1441 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     1501 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     1561 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     1621 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     1681 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     1741 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     1801 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     1861 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     1921 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     1981 ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga
     2041 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt
     2101 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     2161 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     2221 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag
     2281 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2341 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     2401 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2461 catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     2521 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2581 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     2641 ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa
     2701 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     2761 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2821 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2881 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     2941 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3001 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3061 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     3121 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3181 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3241 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3301 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     3361 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3421 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3481 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3541 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3601 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3661 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3721 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3781 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     3841 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     3901 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat
     3961 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4021 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4081 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4141 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4201 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4261 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4321 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4381 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4441 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4501 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4561 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4621 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4681 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4741 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4801 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4861 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4921 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4981 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5041 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5101 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     5161 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5221 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5281 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5341 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5401 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5461 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5521 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5581 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5641 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5701 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5761 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5821 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5881 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5941 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6001 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6061 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6121 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6181 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6241 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6301 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6361 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6421 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc
     6481 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     6541 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     6601 aattgcatga agaatctgct tagg
//
