LOCUS       V025076                 6890 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025076
VERSION     V025076
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..3099
                     /label="DRD1(NM_000794)"
                     /note="DRD1(NM_000794)"
                     /gene="DRD1"
     CDS             3106..3126
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /transl_table=1
                     /translation="ENLYFQS"
                     /label="TEV site"
     CDS             3127..3840
                     /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             3841..3864
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin®, an engineered
                     form of streptavidin"
                     /transl_table=1
                     /translation="WSHPQFEK"
                     /label="Strep-Tag II"
     CDS             3883..3906
                     /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    3978..4033
                     /note="rabbit&nbsp;β-globin polyadenylation signal (Gil and
                     Proudfoot, 1987)"
                     /label="β-globin poly(A) signal"
     primer_bind     complement(4393..4409)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4417..4433
                     /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(4441..4471)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4486..4507
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        4566..4761
                     /note="SV40 early promoter"
                     /label="SV40 promoter"
     polyA_signal    4767..4901
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(5140..5728)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5899..6759)
                     /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(6760..6864)
                     /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 gatgaggact ctgaacacct ctgccatgga cgggactggg
     1801 ctggtggtgg agagggactt ctctgttcgt atcctcactg cctgtttcct gtcgctgctc
     1861 atcctgtcca cgctcctggg gaacacgctg gtctgtgctg ccgttatcag gttccgacac
     1921 ctgcggtcca aggtgaccaa cttctttgtc atctccttgg ctgtgtcaga tctcttggtg
     1981 gccgtcctgg tcatgccctg gaaggcagtg gctgagattg ctggcttctg gccctttggg
     2041 tccttctgta acatctgggt ggcctttgac atcatgtgct ccactgcatc catcctcaac
     2101 ctctgtgtga tcagcgtgga caggtattgg gctatctcca gccctttccg gtatgagaga
     2161 aagatgaccc ccaaggcagc cttcatcctg atcagtgtgg catggacctt gtctgtactc
     2221 atctccttca tcccagtgca gctcagctgg cacaaggcaa aacccacaag cccctctgat
     2281 ggaaatgcca cttccctggc tgagaccata gacaactgtg actccagcct cagcaggaca
     2341 tatgccatct catcctctgt aataagcttt tacatccctg tggccatcat gattgtcacc
     2401 tacaccagga tctacaggat tgctcagaaa caaatacggc gcattgcggc cttggagagg
     2461 gcagcagtcc acgccaagaa ttgccagacc accacaggta atggaaagcc tgtcgaatgt
     2521 tctcaaccgg aaagttcttt taagatgtcc ttcaaaagag aaactaaagt cctgaagact
     2581 ctgtcggtga tcatgggtgt gtttgtgtgc tgttggctac ctttcttcat cttgaactgc
     2641 attttgccct tctgtgggtc tggggagacg cagcccttct gcattgattc caacaccttt
     2701 gacgtgtttg tgtggtttgg gtgggctaat tcatccttga accccatcat ttatgccttt
     2761 aatgctgatt ttcggaaggc attttcaacc ctcttaggat gctacagact ttgccctgcg
     2821 acgaataatg ccatagagac ggtgagtatc aataacaatg gggccgcgat gttttccagc
     2881 catcatgagc cacgaggctc catctccaag gagtgcaatc tggtttacct gatcccacat
     2941 gctgtgggct cctctgagga cctgaaaaag gaggaggcag ctggcatcgc cagacccttg
     3001 gagaagctgt ccccagccct atcagtcata ttggactatg acactgacgt ctctctggag
     3061 aagatccaac ccatcacaca aaacggtcag cacccaaccg aattcgaaaa cctgtacttc
     3121 caatccgtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     3181 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     3241 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     3301 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     3361 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     3421 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     3481 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     3541 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     3601 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     3661 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     3721 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     3781 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     3841 tggagccacc cgcagttcga gaagggcggc ggcagcagcg cctggagcca cccgcagttc
     3901 gagaagtgaa gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     3961 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     4021 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     4081 gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggtgg
     4141 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga
     4201 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta
     4261 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta
     4321 ctcccagtca tagctgtccc tcttctctta tgaagatccc tcgacctgca gcccaagctt
     4381 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4441 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4501 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg
     4561 gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4621 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4681 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4741 ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat
     4801 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4861 gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg
     4921 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4981 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5041 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5101 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5161 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5221 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5281 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     5341 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5401 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5461 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5521 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5581 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5641 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5701 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5761 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5821 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5881 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5941 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6001 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6061 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6121 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6181 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6241 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6301 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6361 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6421 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6481 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6541 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6601 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6661 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6721 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6781 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6841 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
