LOCUS       V025193                 6503 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025193
VERSION     V025193
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        4..383
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        386..661
                     /label="chicken β-actin promoter"
     intron          662..1674
                     /note="chimera between introns from chicken β-actin and
                     rabbit β-globin"
                     /label="chimeric intron"
     regulatory      1681..1690
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             1690..1755
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1762..2712
                     /label="OR51Q1(NM_001004757)"
                     /note="OR51Q1(NM_001004757)"
                     /gene="OR51Q1"
     CDS             2719..2739
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /transl_table=1
                     /translation="ENLYFQS"
                     /label="TEV site"
     CDS             2740..3453
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             3454..3477
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin®, an engineered
                     form of streptavidin"
                     /transl_table=1
                     /translation="WSHPQFEK"
                     /label="Strep-Tag II"
     CDS             3496..3519
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin®, an engineered
                     form of streptavidin"
                     /transl_table=1
                     /translation="WSHPQFEK"
                     /label="Strep-Tag II"
     polyA_signal    3591..3646
                     /note="rabbit&nbsp;β-globin polyadenylation signal (Gil and
                     Proudfoot, 1987)"
                     /label="β-globin poly(A) signal"
     primer_bind     complement(4006..4022)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4030..4046
                     /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(4054..4084)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4099..4120
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        4179..4374
                     /note="SV40 early promoter"
                     /label="SV40 promoter"
     polyA_signal    4380..4514
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(4753..5341)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5512..6372)
                     /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(6373..6477)
                     /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 tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gtgcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 gccaccatgg actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag
     1741 gatgacgatg acaagctcga gatgtcccag gtgactaaca ccacacaaga aggcatctac
     1801 ttcatcctca cggacatccc tggatttgag gcctcccaca tctggatctc catccccgtc
     1861 tgctgtctct acaccatctc catcatgggc aataccacca tcctcactgt cattcgcaca
     1921 gagccatctg tccaccagcg catgtatctg tttctctcca tgctggccct gacggacctg
     1981 ggtctcaccc tcaccaccct acccacagtc atgcagcttc tctggttcaa cgttcgtaga
     2041 atcagctctg aggcctgttt tgctcagttt ttcttccttc atggattctc ctttatggag
     2101 tcttctgtcc tcctggctat gtccgttgac tgctatgtgg ccatctgctg tcccctccat
     2161 tatgcctcca tcctcaccaa tgaagtcatt ggtagaactg ggttagccat catttgctgc
     2221 tgtgttctgg cggttcttcc ctcccttttc ttactcaagc gactgccttt ctgccactcc
     2281 caccttctct ctcgctccta ttgcctccac caggatatga tccgcctggt ctgtgctgac
     2341 atcaggctca acagctggta tggatttgct cttgccttgc tcattattat cgtggatcct
     2401 ctgctcattg tgatctccta tacacttatt ctgaaaaata tcttgggcac agccacctgg
     2461 gctgagcgac tccgtgccct caataactgc ctgtcccaca ttctagctgt cctggtcctc
     2521 tacattccca tggttggtgt atctatgact catcgctttg ccaagcatgc ctctccactg
     2581 gtccatgtta tcatggccaa tatctacctg ctggcacccc cggtgatgaa ccccatcatt
     2641 tacagtgtaa agaacaagca gatccaatgg ggaatgttaa atttcctttc cctcaaaaat
     2701 atgcattcaa gagaattcga aaacctgtac ttccaatccg tgagcaaggg cgaggagctg
     2761 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     2821 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     2881 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     2941 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     3001 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     3061 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     3121 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     3181 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     3241 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     3301 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     3361 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     3421 gggatcactc tcggcatgga cgagctgtac aagtggagcc acccgcagtt cgagaagggc
     3481 ggcggcagca gcgcctggag ccacccgcag ttcgagaagt gaagatcttt ttccctctgc
     3541 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
     3601 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     3661 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
     3721 tgcccatatg ctggctgcca tgaacaaagg tggctataaa gaggtcatca gtatatgaaa
     3781 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt
     3841 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt
     3901 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc
     3961 ttatgaagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc
     4021 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4081 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4141 cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc
     4201 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4261 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4321 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta
     4381 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4441 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4501 atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4561 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4621 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4681 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4741 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4801 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4861 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4921 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     4981 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5041 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5101 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5161 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5221 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5281 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5341 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5401 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5461 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5521 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5581 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5641 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5701 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5761 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5821 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5881 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5941 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6001 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6061 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6121 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6181 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6241 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6301 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6361 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6421 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6481 atttccccga aaagtgccac ctg
//
