LOCUS       V024977                 6791 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024977
VERSION     V024977
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..3000
                     /label="ADRB2(NM_000024)"
                     /note="ADRB2(NM_000024)"
                     /gene="ADRB2"
     CDS             3007..3027
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /transl_table=1
                     /translation="ENLYFQS"
                     /label="TEV site"
     CDS             3028..3741
                     /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             3742..3765
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin®, an engineered
                     form of streptavidin"
                     /transl_table=1
                     /translation="WSHPQFEK"
                     /label="Strep-Tag II"
     CDS             3784..3807
                     /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    3879..3934
                     /note="rabbit&nbsp;β-globin polyadenylation signal (Gil and
                     Proudfoot, 1987)"
                     /label="β-globin poly(A) signal"
     primer_bind     complement(4294..4310)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4318..4334
                     /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(4342..4372)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4387..4408
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        4467..4662
                     /note="SV40 early promoter"
                     /label="SV40 promoter"
     polyA_signal    4668..4802
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(5041..5629)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5800..6660)
                     /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(6661..6765)
                     /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 gatggggcaa cccgggaacg gcagcgcctt cttgctggca
     1801 cccaatggaa gccatgcgcc ggaccacgac gtcacgcagg aaagggacga ggtgtgggtg
     1861 gtgggcatgg gcatcgtcat gtctctcatc gtcctggcca tcgtgtttgg caatgtgctg
     1921 gtcatcacag ccattgccaa gttcgagcgt ctgcagacgg tcaccaacta cttcatcact
     1981 tcactggcct gtgctgatct ggtcatgggc ctggcagtgg tgccctttgg ggccgcccat
     2041 attcttatga aaatgtggac ttttggcaac ttctggtgcg agttttggac ttccattgat
     2101 gtgctgtgcg tcacggccag cattgagacc ctgtgcgtga tcgcagtgga tcgctacttt
     2161 gccattactt cacctttcaa gtaccagagc ctgctgacca agaataaggc ccgggtgatc
     2221 attctgatgg tgtggattgt gtcaggcctt acctccttct tgcccattca gatgcactgg
     2281 taccgggcca cccaccagga agccatcaac tgctatgcca atgagacctg ctgtgacttc
     2341 ttcacgaacc aagcctatgc cattgcctct tccatcgtgt ccttctacgt tcccctggtg
     2401 atcatggtct tcgtctactc cagggtcttt caggaggcca aaaggcagct ccagaagatt
     2461 gacaaatctg agggccgctt ccatgtccag aaccttagcc aggtggagca ggatgggcgg
     2521 acggggcatg gactccgcag atcttccaag ttctgcttga aggagcacaa agccctcaag
     2581 acgttaggca tcatcatggg cactttcacc ctctgctggc tgcccttctt catcgttaac
     2641 attgtgcatg tgatccagga taacctcatc cgtaaggaag tttacatcct cctaaattgg
     2701 ataggctatg tcaattctgg tttcaatccc cttatctact gccggagccc agatttcagg
     2761 attgccttcc aggagcttct gtgcctgcgc aggtcttctt tgaaggccta tgggaatggc
     2821 tactccagca acggcaacac aggggagcag agtggatatc acgtggaaca ggagaaagaa
     2881 aataaactgc tgtgtgaaga cctcccaggc acggaagact ttgtgggcca tcaaggtact
     2941 gtgcctagcg ataacattga ttcacaaggg aggaattgta gtacaaatga ctcactgctg
     3001 gaattcgaaa acctgtactt ccaatccgtg agcaagggcg aggagctgtt caccggggtg
     3061 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     3121 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     3181 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     3241 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3301 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3361 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3421 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3481 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     3541 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3601 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     3661 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3721 ggcatggacg agctgtacaa gtggagccac ccgcagttcg agaagggcgg cggcagcagc
     3781 gcctggagcc acccgcagtt cgagaagtga agatcttttt ccctctgcca aaaattatgg
     3841 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     3901 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     3961 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     4021 ggctgccatg aacaaaggtg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     4081 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
     4141 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     4201 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atgaagatcc
     4261 ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4321 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4381 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4441 gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc
     4501 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4561 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4621 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg
     4681 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4741 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     4801 tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4861 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4921 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4981 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5041 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5101 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5161 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5221 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5281 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5341 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5401 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5461 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     5521 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5581 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5641 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5701 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5761 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5821 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5881 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5941 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6001 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6061 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6121 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6181 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6241 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6301 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6361 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6421 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6481 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6541 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6601 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6661 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6721 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6781 agtgccacct g
//
