LOCUS       V025172                 6938 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025172
VERSION     V025172
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..3147
                     /label="ADRA2C(NM_000683)"
                     /note="ADRA2C(NM_000683)"
                     /gene="ADRA2C"
     CDS             3154..3174
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /transl_table=1
                     /translation="ENLYFQS"
                     /label="TEV site"
     CDS             3175..3888
                     /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             3889..3912
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin®, an engineered
                     form of streptavidin"
                     /transl_table=1
                     /translation="WSHPQFEK"
                     /label="Strep-Tag II"
     CDS             3931..3954
                     /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    4026..4081
                     /note="rabbit&nbsp;β-globin polyadenylation signal (Gil and
                     Proudfoot, 1987)"
                     /label="β-globin poly(A) signal"
     primer_bind     complement(4441..4457)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4465..4481
                     /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(4489..4519)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4534..4555
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        4614..4809
                     /note="SV40 early promoter"
                     /label="SV40 promoter"
     polyA_signal    4815..4949
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(5188..5776)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5947..6807)
                     /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(6808..6912)
                     /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 gatggcgtcc ccggcgctgg cggcggcgct ggcggtggcg
     1801 gcagcggcgg gccccaatgc gagcggcgcg ggcgagaggg gcagcggcgg ggttgccaat
     1861 gcctcggggg cttcctgggg gccgccgcgc ggccagtact cggcgggcgc ggtggcaggg
     1921 ctggctgccg tggtgggctt cctcatcgtc ttcaccgtgg tgggcaacgt gctggtggtg
     1981 atcgccgtgc tgaccagccg ggcgctgcgc gcgccacaga acctcttcct ggtgtcgctg
     2041 gcctcggccg acatcctggt ggccacgctg gtcatgccct tctcgttggc caacgagctc
     2101 atggcctact ggtacttcgg gcaggtgtgg tgcggcgtgt acctggcgct cgatgtgctg
     2161 ttttgcacct cgtcgatcgt gcatctgtgt gccatcagcc tggaccgcta ctggtcggtg
     2221 acgcaggccg tcgagtacaa cctgaagcgc acaccacgcc gcgtcaaggc caccatcgtg
     2281 gccgtgtggc tcatctcggc cgtcatctcc ttcccgccgc tggtctcgct ctaccgccag
     2341 cccgacggcg ccgcctaccc gcagtgcggc ctcaacgacg agacctggta catcctgtcc
     2401 tcctgcatcg gctccttctt cgcgccctgc ctcatcatgg gcctggtcta cgcgcgcatc
     2461 taccgagtgg ccaagctgcg cacgcgcacg ctcagcgaga agcgcgcccc cgtgggcccc
     2521 gacggtgcgt ccccgactac cgaaaacggg ctgggcgcgg cggcaggcgc aggcgagaac
     2581 gggcactgcg cgcccccgcc cgccgacgtg gagccggacg agagcagcgc agcggccgag
     2641 aggcggcggc gccggggcgc gttgcggcgg ggcgggcggc ggcgagcggg cgcggagggg
     2701 ggcgcgggcg gtgcggacgg gcagggggcg gggccggggg cggctgagtc gggggcgctg
     2761 accgcctcca ggtccccggg gcccggtggc cgcctgtcgc gcgccagctc gcgctccgtc
     2821 gagttcttcc tgtcgcgccg gcgccgggcg cgcagcagcg tgtgccgccg caaggtggcc
     2881 caggcgcgcg agaagcgctt cacctttgtg ctggctgtgg tcatgggcgt gttcgtgctc
     2941 tgctggttcc ccttcttctt cagctacagc ctgtacggca tctgccgcga ggcctgccag
     3001 gtgcccggcc cgctcttcaa gttcttcttc tggatcggct actgcaacag ctcgctcaac
     3061 ccggtcatct acacggtctt caaccaggat ttccggcgat cctttaagca catcctcttc
     3121 cgacggagga gaaggggctt caggcaggaa ttcgaaaacc tgtacttcca atccgtgagc
     3181 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     3241 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     3301 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     3361 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     3421 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     3481 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     3541 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     3601 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     3661 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     3721 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     3781 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     3841 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtg gagccacccg
     3901 cagttcgaga agggcggcgg cagcagcgcc tggagccacc cgcagttcga gaagtgaaga
     3961 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
     4021 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     4081 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
     4141 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggtggct ataaagaggt
     4201 catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact
     4261 tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa
     4321 ttttccttac atgttttact agccagattt ttcctcctct cctgactact cccagtcata
     4381 gctgtccctc ttctcttatg aagatccctc gacctgcagc ccaagcttgg cgtaatcatg
     4441 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4501 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4561 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tccgcatctc
     4621 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4681 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4741 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4801 ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4861 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4921 tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg
     4981 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5041 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5101 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5161 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5221 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5281 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5341 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     5401 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5461 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5521 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5581 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     5641 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5701 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5761 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5821 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5881 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5941 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6001 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6061 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6121 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6181 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6241 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6301 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6361 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6421 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6481 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6541 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6601 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6661 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6721 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6781 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6841 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6901 aggggttccg cgcacatttc cccgaaaagt gccacctg
//
