LOCUS       V024920                 6497 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024920
VERSION     V024920
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..2706
                     /label="OR56A3(NM_001003443)"
                     /note="OR56A3(NM_001003443)"
                     /gene="OR56A3"
     CDS             2713..2733
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /transl_table=1
                     /translation="ENLYFQS"
                     /label="TEV site"
     CDS             2734..3447
                     /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             3448..3471
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin®, an engineered
                     form of streptavidin"
                     /transl_table=1
                     /translation="WSHPQFEK"
                     /label="Strep-Tag II"
     CDS             3490..3513
                     /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    3585..3640
                     /note="rabbit&nbsp;β-globin polyadenylation signal (Gil and
                     Proudfoot, 1987)"
                     /label="β-globin poly(A) signal"
     primer_bind     complement(4000..4016)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4024..4040
                     /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(4048..4078)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4093..4114
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        4173..4368
                     /note="SV40 early promoter"
                     /label="SV40 promoter"
     polyA_signal    4374..4508
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(4747..5335)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5506..6366)
                     /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(6367..6471)
                     /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 gatgacaaca caccgaaatg acaccctctc cactgaagct
     1801 tcagacttcc tcttgaattg ttttgtcaga tcccccagct ggcagcactg gctgtccctg
     1861 cccctcagcc tccttttcct cttggccgta ggggccaaca ccaccctcct gatgaccatc
     1921 tggctggagg cctctctgca ccagcccctg tactacctgc tcagcctcct ctccctgctg
     1981 gacatcgtgc tctgcctcac tgtcatcccc aaggtcctga ccatcttctg gtttgacctc
     2041 aggcccatca gcttccctgc ctgcttcctc cagatgtaca tcatgaattg tttcctagcc
     2101 atggagtctt gcacattcat ggtcatggcc tatgatcgtt atgtagccat ctgccaccca
     2161 ctgagatatc catcaatcat cactgatcac tttgtagtca aggctgccat gtttattttg
     2221 accagaaatg tgcttatgac tctgcccatc cccatccttt cagcacaact ccgttattgt
     2281 ggaagaaatg tcattgagaa ctgcatctgt gccaatatgt ctgtttccag actctcctgc
     2341 gatgatgtca ccatcaatca cctttaccaa tttgctggag gctggactct gctaggatct
     2401 gacctcatcc ttatcttcct ctcctacacc ttcattctgc gagctgtgct gagactcaag
     2461 gcagagggtg ccgtggcaaa ggccctaagc acatgtggct cccacttcat gctcatcctc
     2521 ttcttcagca ccatccttct ggtttttgtc ctcacacatg tggctaagaa gaaagtctcc
     2581 cctgatgtgc cagtcttgct caatgttctc caccatgtca ttcctgcagc ccttaacccc
     2641 atcatttacg gggtgagaac ccaagaaatt aagcagggaa tgcagaggtt gttgaagaaa
     2701 gggtgcgaat tcgaaaacct gtacttccaa tccgtgagca agggcgagga gctgttcacc
     2761 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     2821 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     2881 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     2941 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     3001 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     3061 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     3121 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     3181 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     3241 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     3301 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     3361 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     3421 actctcggca tggacgagct gtacaagtgg agccacccgc agttcgagaa gggcggcggc
     3481 agcagcgcct ggagccaccc gcagttcgag aagtgaagat ctttttccct ctgccaaaaa
     3541 ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat
     3601 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     3661 ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca
     3721 tatgctggct gccatgaaca aaggtggcta taaagaggtc atcagtatat gaaacagccc
     3781 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     3841 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     3901 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatga
     3961 agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4021 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4081 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4141 ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc
     4201 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4261 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4321 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt
     4381 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4441 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4501 tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4561 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4621 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4681 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4741 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4801 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4861 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4921 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4981 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5041 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5101 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5161 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     5221 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5281 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5341 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5401 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5461 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5521 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5581 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5641 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5701 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5761 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5821 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5881 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5941 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6001 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6061 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6121 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6181 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6241 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6301 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6361 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6421 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6481 ccgaaaagtg ccacctg
//
