LOCUS       V033839                 7819 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033839
VERSION     V033839
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      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             928..954
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             958..984
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             988..1014
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             1030..3339
                     /label="Trim2(NM_001271725)"
                     /note="Trim2(NM_001271725)"
                     /gene="Trim2"
     polyA_signal    3419..3643
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3689..4117
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4131..4460
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4527..5321
                     /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    5495..5616
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5665..5681)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5689..5705
                     /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(5713..5743)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5758..5779
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6067..6652)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6823..7683)
                     /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(7684..7788)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gctggctagc
      901 gtttaaactt aagccaccat ggcggcgtac ccatacgacg tacctgatta tgcaacttat
      961 ccttatgatg tacctgatta tgctagctat ccttatgatg tcccagacta cgcatcagcg
     1021 gaaaagctta tgcccaggag tggccgttat ggaacccagc agcgtgcagg gtcgaggaca
     1081 gccggccccc catgtcagtg gtctaggatg gccagtgagg gcgccagcat cccaagtccc
     1141 gtggtgcgcc agattgacaa gcagtttctg atctgcagta tatgcctgga acggtacaag
     1201 aaccccaagg tcctgccctg tctgcacact ttctgcgaga ggtgcctgca gaactacatc
     1261 ccagcccaca gtttaaccct ctcctgcccg gtgtgtcgcc agacatccat cctgcccgag
     1321 aaaggggtgg ccgcacttca gaacaacttc tttatcacga acctcatgga tgtgctgcag
     1381 cggacgccag gcagcaacgg cgaggactct tccatcctgg agacggtcac cgctgtggct
     1441 gcaggaaagc ctctttcgtg cccaaaccat gatggaaatg tgatggaatt ttactgccag
     1501 tcctgtgaga ccgccatgtg tcgggagtgc acggaggggg agcacgcgga gcaccccaca
     1561 gtcccgctga aggatgtggt ggagcagcac aaggcctcgc tccaggtcca gctggacgca
     1621 gtcaacaaaa gactcccaga aatagattct gctcttcagt tcatctcaga aatcatccat
     1681 cagttaacca accaaaaggc cagcattgtg gatgacatcc actccacctt tgatgagctg
     1741 cagaagactt taaatgttcg caagagcgtg ctgctcatgg agttggaggt caactatggt
     1801 ctcaaacaca aagtgctgca gtcgcagctg gataccctgc tccaaggcca ggaaagcatt
     1861 aagagctgca gcaacttcac ggctcaggcc ctcaaccatg gcacggagac ggaggtgctg
     1921 ctggtgaaga agcagatgag cgagaagctg aacgaactgg cggaccagga cttcccactg
     1981 cacccgcgcg agaacgacca gctggatttt atcgtggaaa cggagggact gaagaagtct
     2041 atccacaacc tggggaccat cctcaccact aacgcggtcg cctctgagac ggtggccacg
     2101 ggggaggggc tgcggcagac catcatcggg cagcccatgt ccgttactat aaccaccaag
     2161 gacaaagacg gagagctgtg caagacaggc aacgcatacc tcaccgctga gctcagcacc
     2221 ccggacggca gcgtggccga cggagagatc ctggacaaca agaacggcac ctacgagttc
     2281 ttgtatactg tgcagaagga gggggacttc accctgtccc tcaggctcta cgaccagcat
     2341 atccgaggca gcccgtttaa gctcaaggtc atccgctctg ccgacgtgtc ccccaccacc
     2401 gagggcgtga agcgacgagt caagtccccg gggagcgggc acgtcaagca gaaggctgtg
     2461 aagagacctg ccagcatgta cagcaccggc aagcgaaaag agaaccccat cgaggacgac
     2521 ctgatcttcc gagtcggtac caaaggaaga aacaaaggag aatttacaaa cctgcagggt
     2581 gtggctgcat ctacaagtgg aaagatatta attgcagata gtaacaacca gtgtgtgcag
     2641 atcttttcca atgatggaca gttcaaaagt cgtttcggca tccgaggacg atcacccgga
     2701 cagctccagc ggcccacggg ggtagcggtg caccccagtg gggacatcat cattgcagat
     2761 tatgacaaca agtgggttag cattttctcc aatgatggga agtttaagac aaaaattgga
     2821 tcaggaaaac tgatggggcc caaaggagtt tctgtggacc gcaatgggca cattattgtg
     2881 gtggacaaca aggcgtgctg tgtgtttatc ttccagccca acgggaaaat agtcaccagg
     2941 tttggtagcc gaggaaacgg ggacaggcag tttgcaggtc cccactttgc agctgtgaat
     3001 agcaataatg agatcatcat tacagatttc cataatcact ctgtcaaggt atttaatcag
     3061 gaaggagaat tcatgttgaa gtttggctcc aatggagaag gaaatgggca gtttaatgca
     3121 ccaacaggtg tggccgtgga ctcaaatggg aacatcattg tggctgactg ggggaacagc
     3181 aggatccagg tttttgatgg gagtggatca tttttgtcct atatcaacac atctgctgac
     3241 ccgctctatg gcccccaggg cctggctctc acttcagatg gccatgttgt ggttgcagac
     3301 tccggaaacc actgtttcaa ggtctaccgg tacttacagt aagaattctg cagatatcca
     3361 gcacagtggc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     3421 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3481 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3541 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3601 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     3661 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3721 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3781 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3841 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3901 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3961 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4021 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4081 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     4141 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4201 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4261 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4321 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4381 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4441 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4501 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4561 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4621 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4681 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     4741 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4801 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4861 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4921 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4981 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5041 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5101 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5161 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5221 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     5281 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     5341 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5401 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5461 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5521 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5581 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5641 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5701 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5761 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5821 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5881 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5941 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6001 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6061 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6121 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6181 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6241 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6301 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6361 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6421 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6481 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6541 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6601 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6661 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6721 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6781 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6841 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6901 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6961 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7021 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7081 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7141 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7201 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7261 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7321 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7381 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7441 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7501 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7561 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7621 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7681 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7741 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7801 aaaagtgcca cctgacgtc
//
