LOCUS       V024919                 6473 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024919
VERSION     V024919
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..2682
                     /label="OR8K5(NM_001004058)"
                     /note="OR8K5(NM_001004058)"
                     /gene="OR8K5"
     CDS             2689..2709
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /transl_table=1
                     /translation="ENLYFQS"
                     /label="TEV site"
     CDS             2710..3423
                     /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             3424..3447
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin®, an engineered
                     form of streptavidin"
                     /transl_table=1
                     /translation="WSHPQFEK"
                     /label="Strep-Tag II"
     CDS             3466..3489
                     /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    3561..3616
                     /note="rabbit&nbsp;β-globin polyadenylation signal (Gil and
                     Proudfoot, 1987)"
                     /label="β-globin poly(A) signal"
     primer_bind     complement(3976..3992)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4000..4016
                     /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(4024..4054)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4069..4090
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        4149..4344
                     /note="SV40 early promoter"
                     /label="SV40 promoter"
     polyA_signal    4350..4484
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(4723..5311)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5482..6342)
                     /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(6343..6447)
                     /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 gatgggccaa cacaatctaa cagtgctaac tgaattcatt
     1801 ctgatggaac tcacaaggcg gcctgagctg cagattcccc tttttggagt cttcctcgtc
     1861 atctacctaa tcacagtggt gggcaaccta actatgatca ttttgaccaa actggactcc
     1921 cacttacata cacctatgta cttttctatc agacatttgg cttttgttga tcttggtaat
     1981 tctactgtca tttgtcccaa ggtgctggca aattttgttg tggatcgaaa tactatttcc
     2041 tattatgcat gtgctgcaca gctggcattc ttccttatgt tcattatcag tgaatttttc
     2101 atcctgtcag ccatggccta tgaccgctat gtggccattt gtaaccctct gctctattat
     2161 gttattatgt ctcagcgact gtgtcatgta ctggtgggca ttcaatatct ctacagcaca
     2221 tttcaggctc tgatgttcac tattaagatt tttacattga ccttctgtgg ctctaatgtc
     2281 atcagtcatt tttactgtga tgatgttcct ttgctaccta tgctttgctc aaatgcacag
     2341 gaaatagaat tgttgagcat actattttct gtatttaatt tgatctcctc ctttctgata
     2401 gtcttagtgt cctacatgtt gattttgtta gctatatgtc aaatgcattc tgcagagggc
     2461 aggaaaaagg ctttctccac atgtggttcc catttgacag tggtggttgt gttctatggg
     2521 tctctactct tcatgtacat gcagcccaat tccactcact tctttgatac tgataaaatg
     2581 gcttctgtgt tttacacttt agtaatcccc atgcttaacc ctttgattta cagcttaaga
     2641 aacgaagagg tgaaaaatgc cttctataag ctctttgaga atgaattcga aaacctgtac
     2701 ttccaatccg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2761 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     2821 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     2881 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     2941 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     3001 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     3061 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     3121 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     3181 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     3241 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3301 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     3361 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     3421 aagtggagcc acccgcagtt cgagaagggc ggcggcagca gcgcctggag ccacccgcag
     3481 ttcgagaagt gaagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct
     3541 tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     3601 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa
     3661 tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg
     3721 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     3781 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     3841 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     3901 ctactcccag tcatagctgt ccctcttctc ttatgaagat ccctcgacct gcagcccaag
     3961 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4021 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4081 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4141 gcggatccgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4201 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4261 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4321 tttggaggcc taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca
     4381 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4441 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat
     4501 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4561 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4621 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4681 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4741 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4801 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4861 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     4921 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4981 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5041 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5101 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5161 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5221 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5281 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5341 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5401 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5461 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5521 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5581 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5641 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5701 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5761 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5821 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5881 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5941 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6001 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6061 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6121 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6181 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6241 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6301 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6361 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6421 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctg
//
