LOCUS       V031605                 9171 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031605
VERSION     V031605
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..3851
                     /label="CARD10(NM_014550)"
                     /note="CARD10(NM_014550)"
                     /gene="CARD10"
     CDS             3858..3923
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3947..4535
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4601..4649
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4851..5279
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5293..5622
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5689..6483
                     /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    6659..6780
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6829..6845)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6853..6869
                     /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(6877..6907)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6922..6943
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7231..7819)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7990..8850)
                     /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(8851..8955)
                     /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 ccaccatgcc gggccgggcg gaggcggggg
      781 aggccgagga ggaggccggg gccggctcgg ggtctgaggc ggaggaggac gcgctgtggg
      841 agcgaatcga gggcgtccgg catcggctgg ctcgcgccct gaacccggcc aagctcacgc
      901 cgtatctgcg ccagtgccgg gtcatcgacg agcaggacga ggaggaggtg ctgagcacct
      961 accgcttccc gtgccgcgtc aaccgcaccg ggcgcctgat ggacatcttg cgctgccgtg
     1021 gcaagagggg ctatgaggcc ttcctggaag ccctggagtt ctactacccc gaacacttca
     1081 cgctgctcac gggccaggaa cccgcccagc gctgctccat gatcctcgat gaggaggggc
     1141 ctgagggcct gacccaattc ttgatgacag aggtgcgacg gctgcgggaa gctcgcaaga
     1201 gccagctgca gcgggagcag caactgcagg cccggggccg ggtgctcgag gaggagcggg
     1261 cagggctgga gcagcggctg cgggaccagc agcaggctca ggagcgctgt caacggctgc
     1321 gggaggactg ggaggcgggc agcctggagc tgctgcggct caaggatgag aactacatga
     1381 tcgccatgcg cctggcacag ctcagtgagg agaagaactc ggctgtactt cgcagccgtg
     1441 acctgcagct ggcggtggat cagctcaagc tcaaagtgag tcggctggag gaagagtgtg
     1501 cactgcttcg aagggccagg ggcccgcccc ctggggcaga ggagaaggag aaggagaagg
     1561 agaaggagaa ggagccagac aatgtggacc ttgtctctga gctgcgtgct gagaaccagc
     1621 ggctgacggc gtcactgcgg gagttgcagg agggcctgca gcaggaggcg agccggccgg
     1681 gggccccggg ctccgagcgc atcctgctgg acatcctaga gcatgactgg cgggaggcgc
     1741 aggacagcag gcaggagctg tgccagaagc tgcatgccgt gcagggggag ctgcagtggg
     1801 ccgaggagct gcgcgaccag tacctgcagg agatggaaga cctgcggctc aagcaccgca
     1861 cgctgcagaa ggactgtgac ctgtacaagc accgcatggc cactgtcctg gcccaactgg
     1921 aggagattga gaaggagcga gaccaggcca tccagagccg tgaccggatc cagttgcagt
     1981 actcacagag cctcatcgag aaggaccagt accgcaagca ggtgcggggc ctggaggcgg
     2041 agcgggatga gctgctgaca acgctcacca gcctggaggg caccaaggct ctgctggagg
     2101 ttcagctgca gcgggcccag ggtggcacct gcctcaaggc ctgtgcctcc tcccattccc
     2161 tgtgctccaa cctcagcagc acttggagcc tgagcgagtt cccctcccct ctgggaggcc
     2221 cagaagcaac tggggaggca gctgtcatgg ggggacctga gcctcacaac tcggaggaag
     2281 ccacagacag tgaaaaggag atcaatcggc tctccatcct gcccttcccc cccagtgccg
     2341 gctccatcct ccgccggcag cgtgaggaag accccgcacc ccctaagaga tccttcagca
     2401 gcatgtcaga catcacaggg agtgtgacac ttaagccctg gtcccctggc ctctcttcgt
     2461 cctcatcctc tgacagcgtg tggcctttgg gaaagccgga aggcctcctg gctcggggct
     2521 gtggcctgga cttcctcaac aggtctctgg ctattcgggt gtctggccgg agccccccag
     2581 ggggcccaga gccgcaggac aagggaccag atggactgtc gttttatggg gacagatggt
     2641 ctggggctgt ggtgcgcagg gtgctgtctg ggcctgggtc cgccaggatg gaaccaagag
     2701 agcaaagggt ggaagctgct ggtctggagg gggcgtgcct ggaagccgag gcccagcaga
     2761 gaaccttgct ctggaatcag gggtccacac tcccctccct gatggactcg aaggcctgcc
     2821 agtccttcca cgaggcccta gaagcctggg caaagggacc aggtgccgag cccttctaca
     2881 ttcgtgccaa cctcaccttg cctgagaggg cagatcccca tgccctttgc gtgaaagccc
     2941 aagagatcct tcgactggtg gactcggcat acaagcggag gcaggaatgg ttctgcaccc
     3001 gggttgaccc cctcactctg cgggacctgg accggggcac cgtgcccaat tatcagagag
     3061 cccagcagct cctagaagtt caggagaaat gcctgccctc cagccggcac cgaggccccc
     3121 gcagtaatct gaagaagaga gccctggacc agctgcggct ggtgaggccc aagcccgtgg
     3181 gggcgcctgc aggggactcc ccggatcagc tgctgctgga gccctgtgca gagccggagc
     3241 ggagcctcag accctacagt ttggtgcggc cgctactggt gtctgccctg cggcccgtgg
     3301 tgctgttgcc tgagtgcctg gcgccccggc tcatccgtaa cctgctagac ctgcccagct
     3361 cccggctgga cttccaagtg tgcccagcgg aaagcctctc tggggaggaa ctgtgcccat
     3421 cctcagcgcc tggagccccc aaggctcagc ctgccacccc tgggctaggc agcaggatcc
     3481 gggccatcca ggagtctgtt gggaagaagc actgcctgct ggagctgggt gctcggggtg
     3541 tgcgggagct ggtgcagaac gagatctacc ccatcgtcat ccacgtggag gtgactgaga
     3601 agaatgtccg ggaagtcagg ggtctgctgg gccggccggg ctggcgggac tcagagctgc
     3661 tgcggcagtg ccgtggctca gagcaggtgc tctgggggct gccctgctcc tgggtgcagg
     3721 tgcccgccca tgagtgggga cacgcagagg agctggccaa ggtggtgcgc ggccgcatcc
     3781 tgcaggagca ggcccgcctc gtgtgggtgg agtgcggcag cagcagaggc tgccccagca
     3841 gcagtgaggc cggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg
     3901 actacaagga tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga
     3961 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     4021 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     4081 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     4141 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     4201 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     4261 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     4321 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     4381 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     4441 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     4501 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa
     4561 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     4621 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     4681 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     4741 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     4801 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     4861 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4921 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4981 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     5041 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5101 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5161 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5221 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     5281 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     5341 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     5401 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     5461 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5521 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     5581 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     5641 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     5701 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5761 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5821 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     5881 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5941 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     6001 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     6061 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     6121 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     6181 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     6241 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     6301 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     6361 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     6421 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     6481 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     6541 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     6601 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     6661 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     6721 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     6781 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     6841 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6901 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6961 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     7021 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     7081 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     7141 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     7201 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     7261 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     7321 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7381 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     7441 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7501 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7561 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7621 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     7681 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7741 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7801 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7861 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7921 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7981 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     8041 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     8101 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     8161 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     8221 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     8281 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     8341 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8401 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8461 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8521 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8581 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8641 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8701 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8761 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8821 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8881 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8941 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     9001 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     9061 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     9121 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
