LOCUS       V025106                 6491 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025106
VERSION     V025106
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..2700
                     /label="OR51A2(NM_001004748)"
                     /note="OR51A2(NM_001004748)"
                     /gene="OR51A2"
     CDS             2707..2727
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /transl_table=1
                     /translation="ENLYFQS"
                     /label="TEV site"
     CDS             2728..3441
                     /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             3442..3465
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin®, an engineered
                     form of streptavidin"
                     /transl_table=1
                     /translation="WSHPQFEK"
                     /label="Strep-Tag II"
     CDS             3484..3507
                     /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    3579..3634
                     /note="rabbit&nbsp;β-globin polyadenylation signal (Gil and
                     Proudfoot, 1987)"
                     /label="β-globin poly(A) signal"
     primer_bind     complement(3994..4010)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4018..4034
                     /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(4042..4072)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4087..4108
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        4167..4362
                     /note="SV40 early promoter"
                     /label="SV40 promoter"
     polyA_signal    4368..4502
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(4741..5329)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5500..6360)
                     /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(6361..6465)
                     /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 gatgtccatt atcaacacat catatgttga aatcaccacc
     1801 ttcttcttgg ttgggatgcc agggctagaa tatgcacaca tctggatctc tatccccatc
     1861 tgcagcatgt atcttattgc tattctagga aatggcacca ttctttttat catcaagaca
     1921 gagccctcct tgcatgggcc catgtactat tttctttcca tgttggctat gtcagacttg
     1981 ggtttgtctt tatcatctct gcccactgtg ttaagcatct tcctgttcaa tgcccctgaa
     2041 acttcttcta gtgcctgctt tgcccaggaa ttcttcattc atggattctc agtactggag
     2101 tcctcagtcc tcctgatcat gtcatttgat agattcctag ccatccacaa tcctctgaga
     2161 tacacctcaa tcctgacaac tgtcagagtt gcccaaatag ggatagtatt ctcctttaag
     2221 agcatgctcc tggttcttcc cttccctttc actttaagaa gcttgagata ttgcaagaaa
     2281 aaccaattat cccattccta ctgtctccac caggatgtca tgaagttggc ctgttctgac
     2341 aacagaattg atgttatcta tggctttttt ggagcactct gccttatggt agactttatt
     2401 ctcattgctg tgtcttacac cctgatcctc aagactgtac cgggaattgc atccaaaaag
     2461 gaggagctta aggctctcaa tacttgtgtt tcacacatct gtgcagtgat catcttctac
     2521 ctgcccatca tcaacctggc cgttgtccac cgctttgccg ggcatgtctc tcccctcatt
     2581 aatgttctca tggcaaatgt tctcctactt gtacctccgc tgatgaaacc aattgtttat
     2641 tgtgtaaaaa ctaaacagat tagagtgaga gttgtagcaa aattgtgtca atggaagatt
     2701 gaattcgaaa acctgtactt ccaatccgtg agcaagggcg aggagctgtt caccggggtg
     2761 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     2821 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2881 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     2941 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3001 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3061 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3121 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3181 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     3241 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3301 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     3361 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3421 ggcatggacg agctgtacaa gtggagccac ccgcagttcg agaagggcgg cggcagcagc
     3481 gcctggagcc acccgcagtt cgagaagtga agatcttttt ccctctgcca aaaattatgg
     3541 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     3601 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     3661 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     3721 ggctgccatg aacaaaggtg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     3781 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
     3841 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     3901 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atgaagatcc
     3961 ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4021 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4081 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4141 gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc
     4201 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4261 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4321 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg
     4381 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4441 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     4501 tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4561 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4621 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4681 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4741 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4801 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4861 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4921 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4981 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5041 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5101 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5161 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     5221 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5281 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5341 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5401 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5461 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5521 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5581 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5641 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5701 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5761 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5821 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5881 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5941 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6001 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6061 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6121 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6181 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6241 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6301 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6361 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6421 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6481 agtgccacct g
//
