LOCUS       V031379                 8657 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031379
VERSION     V031379
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      895..904
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             904..927
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             1093..4155
                     /label="Card10(NM_130859)"
                     /note="Card10(NM_130859)"
                     /gene="Card10"
     promoter        complement(4209..4227)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     polyA_signal    4253..4477
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4523..4951
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4965..5295
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5362..6156
                     /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    6330..6451
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6500..6516)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6524..6540
                     /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(6548..6578)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6593..6614
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6902..7490)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7661..8521)
                     /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(8522..8626)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttgccacc
      901 atggactaca aggacgacga tgataagggt accgagctcg gatccatgtt gggtggcccg
      961 ggaggccgcc gggctggggg cggagccaat gaccccgaag gcaccgcccc gtcgcgcctc
     1021 cggagccgtc ggccgagcac agagccggga gctcagggca gctgcagcgc ccgtggctcg
     1081 gaggacgcgg ccatgcaggg ccgggcggac gcgggcgagg ctgacgagga ggctggggcg
     1141 ggttcggggt cggaggccga agaggacgcg ctgtgggagc ggatcgaggg cgtccggcac
     1201 cgactgacgc gcgccctgaa tccggccaag ctcacgccgt atctgcgcca gtgccgcgtc
     1261 ctggacgaac aggacgagga ggaggtgctc agcacctacc gtttcccgtg ccgcgctaac
     1321 cgtaccgggc gcctcataga cattttacgc tgccggggca agaggggctt cgaggccttc
     1381 ctggaagccc tggaattcta ctacccagaa cacttcacgc tgctcactgg ccaggaacct
     1441 gcccaacgct gctccatgat ccttgatgag gagggtcccg agggcctgac tcagttcttg
     1501 atgactgagg tccggaggct tcgggaagct cggaagagcc aattgcaccg agagcagcaa
     1561 ctgcaggccc ggggccgggc cctggaggag gagcgggcag ggctagagca gaggcttcgg
     1621 gagcagcagc aggcacagga gcgctgccag cgactgcggg aggactggga ggctggcagc
     1681 ctggagctgc tgcgcctcaa ggatgagaac tacatgatcg ccatgcgcct ggctcagctc
     1741 agcgaggaga agaattcggc tgtgctgcgc agccgagacc tgcagctggc ggtggaccag
     1801 ctcaagctca aggtcagccg gctggaggaa gagtgtgcgc tgctgaggag ggccaggggc
     1861 ccaccgcctg gggctgagga gaaggagagg gagccagatg gcgctgatct tctctctgag
     1921 cttcgtgcag agaaccagcg gctgacggct tcactgcagg agcttcagga aggtcttcag
     1981 caggagatga gccggccagg agctgcaggc tctgaacgta tcctactgga catactggag
     2041 cacgactgga gggaggcgca ggacagcagg caggagctgt gtcagaagct tcatgctgtg
     2101 cagggcgagc tacagtgggc agaggagctg cgggacaagt acctgcaaga gatggaggat
     2161 ctgcggctca aacatcgaac gctgctgaag gactgtgacc tctataagca ccgcatggcc
     2221 actgtgttgg cccagctgga ggagattgag aaagagcggg accaggcaat ccagagccga
     2281 gaccggatcc agctgcagta ttctcagagc ctcattgaga aggaccagta ccgcaagcag
     2341 gtgcgggggc tggaggcgga gcgcgatgag ctgctcacca cagtcaccag cctggagggg
     2401 accaaggcca tgctggaggc acagctgcag cggacacagg ggggttcctg tctcaaggcc
     2461 tgtgcctcct cccactccct gtgctccaac ctcagcagca cctggagcct cagcgagttc
     2521 ccatcgcccc tgggaggccc agaagctaca ggagaagccg ggggatctga gccccacacc
     2581 tcggaggaag ccacagacag cgagaaggag atcaaccggc tctctatcct gcctttccca
     2641 cccagtgctg gttccatact ccgccggcag cgtgaggaag acccggagcc ccctaagagg
     2701 tcttttagca gcatgtcgga tatcacaggg agtgtgaccc ttaagccatg gtcccctggc
     2761 ctctcttcat catcctcgtc tgacagtgtg tggcctttgg gaaagccgga aggcctcctg
     2821 gctcggggct gtggcctgga cttcctcaat aggtctctgg ccatccgagt gtccggctgg
     2881 agtccccccg caggcctgga tccccaggat aagagtccag atagcatgcc gggccttgga
     2941 gacaggtggt ctggggcggt ggtgcgaagg gtgttgtctg ggcctgggtc agccaggaca
     3001 gaacagaaag agccgagggc ggaaggcact ggtctggagg gggcaggcct ggaagcggag
     3061 gcccagcaga ggaccttgcc ctggaaccag agctccaccc tccccttcct gctggactca
     3121 aaggcctgtc attccttcca tgaggccctg gatgcctggg caaagggacc gggtgccgag
     3181 cccttctaca ttcgagccaa cttctccctg cccgagcggt cggatcccca tgccctttgt
     3241 gtgaaagccc aagagatcct gcggttggtg gacccagccc acaagcggag gcaggagtgg
     3301 ttctgtactc gagtggacac cctcacctta cgggacctgg accggggcac cgtgcccaat
     3361 tatcagagag ctcagcagct cctagaagtc caggaaaaat acttgatctc cagcagacac
     3421 cgaagtcctc gaagcaacct gaagaagcga gccctagggt tggtgaggcc caagcctgca
     3481 gggggcactg caggagactc agcagaacag ctccctgcag agccctgctc agagctagaa
     3541 cgaagcctaa aaccctacag cctggtgcga ccgctgttgg tgtctgcctt gaggcctgtg
     3601 gtcctcctgc ctgagtgcct ggcgcctcgc ctcatccgta acctgctgga tctgccaagc
     3661 tccaggctgg atttccaagt gtgcccagca gagagcctct ctggggagga acagtgtacg
     3721 tcctcagctc ccggagcccc caaagcctgg ccggccactg ctgggctagg cagcaggatc
     3781 cgtgccatcc aggagtctgt cgggaagaag cactgcctgt tggagctggg tgcacgggga
     3841 gtgcgggagt tggtgcacag cgaggtctac cccattgtca tccacgtgga ggtgactgag
     3901 aagaatgtcc gtgagatcag gggcctgttg ggccggccgg gctggcggga ttcagaactg
     3961 ctccgtcagt gtcggggctc agaacagtgg ctctggggac tgccgtgttc ctgggtgcag
     4021 gtgcctgcgc atgcgtgggg ccatgcagag gagctggcca aggtggtgcg tggccgaatc
     4081 ttgcaggaac aggctcgcct tgtgtgggtg gagcgaggca gcagcagagg gggcagtggc
     4141 agcagtagtg aggcctgaga tatccatcac actggcggcc gctcgagcat gcatctagag
     4201 ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct
     4261 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     4321 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     4381 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     4441 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     4501 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4561 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4621 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc
     4681 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4741 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4801 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4861 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat
     4921 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     4981 gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc aaagcatgca tctcaattag
     5041 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5101 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5161 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5221 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5281 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     5341 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     5401 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     5461 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5521 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     5581 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5641 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5701 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5761 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5821 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5881 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5941 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     6001 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     6061 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     6121 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     6181 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     6241 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     6301 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     6361 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     6421 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     6481 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6541 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6601 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6661 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6721 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6781 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6841 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6901 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6961 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7021 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7081 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7141 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7201 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7261 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7321 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     7381 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7441 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7501 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7561 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7621 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7681 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7741 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7801 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7861 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7921 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7981 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     8041 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8101 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8161 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8221 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8281 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8341 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8401 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8461 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8521 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8581 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8641 aagtgccacc tgacgtc
//
