LOCUS       V022516                 9279 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022516
VERSION     V022516
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             744..770
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             774..800
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             804..830
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             843..3953
                     /label="CARD6(NM_032587)"
                     /note="CARD6(NM_032587)"
                     /gene="CARD6"
     misc_feature    4055..4643
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4709..4757
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4959..5387
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5401..5730
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5797..6591
                     /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    6767..6888
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6937..6953)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6961..6977
                     /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(6985..7015)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7030..7051
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7339..7927)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8098..8958)
                     /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(8959..9063)
                     /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 gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagc tagcgccacc atgtacccat acgacgtacc tgattatgca acttatcctt
      781 atgatgtacc tgattatgct agctatcctt atgatgtccc agactacgca aagcttggta
      841 ccatggctac cgagagtact ccctcagaga tcatagaaag agaaagaaaa aagttgcttg
      901 aaatccttca acatgatcct gattctatct tagacacgtt aacttctcgg aggctgattt
      961 ctgaggaaga gtatgagact ctggagaatg ttacagatct cctgaagaaa agtcggaagc
     1021 tgttaatttt ggtacagaaa aagggagagg cgacctgtca gcattttctc aagtgtttat
     1081 ttagtacttt tccacagtca gctgccattt gcggcttaag gcatgaagtt ttaaaacatg
     1141 agaatacagt acctcctcaa tctatggggg caagcagtaa ttcagaagat gctttttctc
     1201 ctggaataaa acagcctgaa gcccctgaga tcacagtgtt cttcagtgag aaggaacact
     1261 tggatttgga aacctctgag tttttcaggg acaagaaaac tagttatagg gaaacagctt
     1321 tgtctgccag gaagaatgag aaggaatatg acacaccaga agtcacatta tcatattcag
     1381 ttgagaaagt tggatgtgaa gttccagcaa ctattacata tataaaagat ggacagagat
     1441 atgaggagct agatgattct ttatacttag gaaaagagga atatctagga tctgttgaca
     1501 cccctgaaga tgcagaagcc actgtggaag aggaggttta tgatgaccca gagcacgttg
     1561 gatatgatgg tgaagaggac ttcgagaatt cagaaaccac agagttctct ggtgaagaac
     1621 caagttatga gggatcagaa accagccttt cattggagga ggaacaggag aaaagtatag
     1681 aagaaagaaa aaaggtgttt aaagatgtcc tgttatgttt gaacatggat agaagcagaa
     1741 aggttctgcc agattttgtt aaacaattct ccttagatcg aggatgtaag tggacccctg
     1801 agagtccagg agacttagcc tggaatttcc tgatgaaagt tcaagcacga gatgtgacgg
     1861 ctagggattc aatcctcagt cacaaggttc tggatgaaga tagcaaggag gatttgctgg
     1921 ctggagtgga gaatttggaa attcgagaca tacaaaccat taatcccctt gacgtgcttt
     1981 gtgccaccat gctgtgttca gatagctctt tgcaacgcca agtcatgtca aacatgtatc
     2041 agtgccagtt tgctcttccc ctgctactgc cagatgcaga aaacaacaaa agcatcttaa
     2101 tgctgggggc catgaaagac attgtgaaga agcagtcaac acagttttca ggggggccta
     2161 cagaggatac agaaaagttt ctgactctca tgaagatgcc tgtcatctct tttgtgcgtc
     2221 taggatactg tagcttctct aagtccagaa tcctcaacac acttctcagc cctgcccagt
     2281 tgaaattaca caaaatcttt cttcatcaag atttgcctct tttggtgctt ccccggcaaa
     2341 tctctgatgg cctggttgag ataacatggt gttttcctga tagcgatgat agaaaggaaa
     2401 accccttttt ccaaaagcct gttgctctgg ctaatctccg tggaaatcta gaaagctttt
     2461 ggactcagtt tggttttttg atggaagttt cttcagctgt gttttttttc actgactgtt
     2521 taggtgagaa ggaatgggac ttgctaatgt ttttaggaga ggctgccatt gaaagatgct
     2581 actttgttct cagttcccaa gccagggaga gtgaagaggc tcaaattttt cagaggatac
     2641 tgaacttgaa gccagcacag ctactgtttt gggagagggg agatgctggg gatagaagga
     2701 agaacatgga gggccttcaa gctgccctcc aggaagtgat gttctcttct tgcctcagat
     2761 gtgtgtctgt ggaggatatg gccgccctgg ccagggagct ggggattcag gtagatgaag
     2821 actttgaaaa cactcagaga attcaagttt cctctggaga aaacatggct gggacagctg
     2881 aaggtgaggg tcagcaaaga cacagtcagc taaaaagctc atctaaaagc caggctctaa
     2941 tgccaattca agagcctggg actcaatgtg agctcagcca gaatcttcag aatctctatg
     3001 gtaccccagt attcaggcct gttctagaga actcctggct ctttccaacc agaattggag
     3061 gtaactttaa ccatgtttcc ttgaaagcct cctgggttat gggccgcccc tttgggtcag
     3121 agcagaggcc taagtggttc catcctttgc cttttcagaa tgcaggggcc cagggccgag
     3181 gtaaaagttt tggtattcaa tccttccatc cccagatatt ttattcaggt gaaagattca
     3241 tgaaattttc cagagttgct cggggatgtc actcgaatgg aacatttggg agactgccaa
     3301 gacccatttg tcagcatgta caggcctgcc ctgagagacc acaaatgatg ggaactcttg
     3361 aaaggtctag ggcagtagcc tccaagatag gtcactccta ttccctggat tcacagccag
     3421 caagagcagt agggaagcca tggcctcagc aagcttgcac cagggtaaca gagttaactg
     3481 aagcaactgg aaaactgata agaacatccc atattggaaa gcctcaccct cagtcctttc
     3541 aaccagcagc agccacacaa aaactaagac ctgcttctca gcaaggagtc cagatgaaga
     3601 cacaaggtgg ggcttcaaat ccagctctcc aaatagggtc ccatcccatg tgcaagagct
     3661 ctcagttcaa atccgatcag tccaacccat ccacagtcaa acactcccag cctaaaccct
     3721 tccattctgt gccctctcaa cctaaatcct ctcagacaaa atcctgtcag tcccagccct
     3781 cccaaactaa accttctcca tgcaaatcta ctcagcctaa gccaagccag ccctggcctc
     3841 cccagtctaa gccttctcag cccagacccc ctcaacctaa gtcatcctca accaatcctt
     3901 cacaagctaa ggcacaccac tcaaaagcag ggcagaagag gggagggaag cattaagaat
     3961 tctgcagata tccagcacag tggcggccgc tcgagtctag aggcccctta agggttcgat
     4021 ccctaccggt tagtaatgag tttgatatct cgacaatcaa cctctggatt acaaaatttg
     4081 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     4141 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     4201 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     4261 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4321 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     4381 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     4441 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     4501 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     4561 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     4621 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     4681 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     4741 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     4801 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     4861 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     4921 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     4981 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     5041 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     5101 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     5161 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     5221 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     5281 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     5341 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     5401 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5461 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     5521 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5581 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5641 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     5701 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     5761 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5821 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5881 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5941 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     6001 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     6061 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     6121 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     6181 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     6241 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     6301 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     6361 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     6421 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     6481 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     6541 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     6601 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6661 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6721 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     6781 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6841 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     6901 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6961 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     7021 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7081 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7141 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7201 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7261 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7321 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7381 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7441 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7501 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     7561 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7621 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7681 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7741 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     7801 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7861 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7921 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7981 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     8041 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     8101 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8161 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     8221 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8281 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8341 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8401 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8461 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8521 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8581 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8641 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     8701 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8761 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8821 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8881 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8941 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     9001 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     9061 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     9121 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     9181 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     9241 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
