LOCUS       V031524                 7573 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031524
VERSION     V031524
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        4..383
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        385..662
                     /label="chicken β-actin promoter"
     intron          663..1680
                     /note="chimera between introns from chicken β-actin and
                     rabbit β-globin"
                     /label="chimeric intron"
     regulatory      1729..1738
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             1738..1761
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             1786..4560
                     /label="RIGI(NM_014314)"
                     /note="RIGI(NM_014314)"
                     /gene="RIGI"
     polyA_signal    4660..4715
                     /note="rabbit&nbsp;β-globin polyadenylation signal (Gil and
                     Proudfoot, 1987)"
                     /label="β-globin poly(A) signal"
     primer_bind     complement(5076..5092)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5100..5116
                     /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(5124..5154)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5169..5190
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        5249..5444
                     /note="SV40 early promoter"
                     /label="SV40 promoter"
     polyA_signal    5450..5584
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(5822..6410)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6581..7441)
                     /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(7442..7546)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggggcccc ccccaggcgg ggcggggcgg ggcgaggggc
      541 ggggcggggc gaggcggaaa ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgt tgcgcgctgc cttccccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaaggggc gcggcggccc ccggagcgcc
     1321 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
     1381 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
     1441 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
     1501 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt
     1561 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
     1621 gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag
     1681 ctcctgggca acgtgctggt tgttgtgctg tctcatcatt ttggcaaagc caccatggat
     1741 tacaaggatg acgacgataa ggaattcgag ctcatcgatg gtaccatgac caccgagcag
     1801 cgacgcagcc tgcaagcctt ccaggattat atccggaaga ccctggaccc tacctacatc
     1861 ctgagctaca tggccccctg gtttagggag gaagaggtgc agtatattca ggctgagaaa
     1921 aacaacaagg gcccaatgga ggctgccaca ctttttctca agttcctgtt ggagctccag
     1981 gaggaaggct ggttccgtgg ctttttggat gccctagacc atgcaggtta ttctggactt
     2041 tatgaagcca ttgaaagttg ggatttcaaa aaaattgaaa agttggagga gtatagatta
     2101 cttttaaaac gtttacaacc agaatttaaa accagaatta tcccaaccga tatcatttct
     2161 gatctgtctg aatgtttaat taatcaggaa tgtgaagaaa ttctacagat ttgctctact
     2221 aaggggatga tggcaggtgc agagaaattg gtggaatgcc ttctcagatc agacaaggaa
     2281 aactggccca aaactttgaa acttgctttg gagaaagaaa ggaacaagtt cagtgaactg
     2341 tggattgtag agaaaggtat aaaagatgtt gaaacagaag atcttgagga taagatggaa
     2401 acttctgaca tacagatttt ctaccaagaa gatccagaat gccagaatct tagtgagaat
     2461 tcatgtccac cttcagaagt gtctgataca aacttgtaca gcccatttaa accaagaaat
     2521 taccaattag agcttgcttt gcctgctatg aaaggaaaaa acacaataat atgtgctcct
     2581 acaggttgtg gaaaaacctt tgtttcactg cttatatgtg aacatcatct taaaaaattc
     2641 ccacaaggac aaaaggggaa agttgtcttt tttgcgaatc agatcccagt gtatgaacag
     2701 cagaaatctg tattctcaaa atactttgaa agacatgggt atagagttac aggcatttct
     2761 ggagcaacag ctgagaatgt cccagtggaa cagattgttg agaacaatga catcatcatt
     2821 ttaactccac agattcttgt gaacaacctt aaaaagggaa cgattccatc actatccatc
     2881 tttactttga tgatatttga tgaatgccac aacactagta aacaacaccc gtacaatatg
     2941 atcatgttta attatctaga tcagaaactt ggaggatctt caggcccact gccccaggtc
     3001 attgggctga ctgcctcggt tggtgttggg gatgccaaaa acacagatga agccttggat
     3061 tatatctgca agctgtgtgc ttctcttgat gcgtcagtga tagcaacagt caaacacaat
     3121 ctggaggaac tggagcaagt tgtttataag ccccagaagt ttttcaggaa agtggaatca
     3181 cggattagcg acaaatttaa atacatcata gctcagctga tgagggacac agagagtctg
     3241 gcaaagagaa tctgcaaaga cctcgaaaac ttatctcaaa ttcaaaatag ggaatttgga
     3301 acacagaaat atgaacaatg gattgttaca gttcagaaag catgcatggt gttccagatg
     3361 ccagacaaag atgaagagag caggatttgt aaagccctgt ttttatacac ttcacatttg
     3421 cggaaatata atgatgccct cattatcagt gagcatgcac gaatgaaaga tgctctggat
     3481 tacttgaaag acttcttcag caatgtccga gcagcaggat tcgatgagat tgagcaagat
     3541 cttactcaga gatttgaaga aaagctgcag gaactagaaa gtgtttccag ggatcccagc
     3601 aatgagaatc ctaaacttga agacctctgc ttcatcttac aagaagagta ccacttaaac
     3661 ccagagacaa taacaattct ctttgtgaaa accagagcac ttgtggacgc tttaaaaaat
     3721 tggattgaag gaaatcctaa actcagtttt ctaaaacctg gcatattgac tggacgtggc
     3781 aaaacaaatc agaacacagg aatgaccctc ccggcacaga agtgtatatt ggatgcattc
     3841 aaagccagtg gagatcacaa tattctgatt gccacctcag ttgctgatga aggcattgac
     3901 attgcacagt gcaatcttgt catcctttat gagtatgtgg gcaatgtcat caaaatgatc
     3961 caaaccagag gcagaggaag agcaagaggt agcaagtgct tccttctgac tagtaatgct
     4021 ggtgtaattg aaaaagaaca aataaacatg tacaaagaaa aaatgatgaa tgactctatt
     4081 ttacgccttc agacatggga cgaagcagta tttagggaaa agattctgca tatacagact
     4141 catgaaaaat tcatcagaga tagtcaagaa aaaccaaaac ctgtacctga taaggaaaat
     4201 aaaaaactgc tctgcagaaa gtgcaaagcc ttggcatgtt acacagctga cgtaagagtg
     4261 atagaggaat gccattacac tgtgcttgga gatgctttta aggaatgctt tgtgagtaga
     4321 ccacatccca agccaaagca gttttcaagt tttgaaaaaa gagcaaagat attctgtgcc
     4381 cgacagaact gcagccatga ctggggaatc catgtgaagt acaagacatt tgagattcca
     4441 gttataaaaa ttgaaagttt tgtggtggag gatattgcaa ctggagttca gacactgtac
     4501 tcgaagtgga aggactttca ttttgagaag ataccatttg atccagcaga aatgtccaaa
     4561 tgactcgagc tagcagatct taattaatta aggatctttt tccctctgcc aaaaattatg
     4621 gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca
     4681 ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa
     4741 atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc
     4801 tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg
     4861 ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt
     4921 gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca
     4981 gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat
     5041 ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5101 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5161 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5221 tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc atagtcccgc
     5281 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5341 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     5401 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta acttgtttat
     5461 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5521 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5581 gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5641 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5701 tcagctcact caaaggcggt aatacggtta tccacagaat cagggataac gcaggaaaga
     5761 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5821 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5881 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5941 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6001 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6061 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6121 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6181 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6241 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6301 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6361 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6421 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6481 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6541 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6601 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6661 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6721 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6781 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6841 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6901 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6961 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7021 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7081 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7141 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7201 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7261 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7321 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7381 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7441 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7501 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7561 aagtgccacc tgg
//
