LOCUS V025057 6668 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025057 VERSION V025057 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..2877 /label="CXCR5(NM_001716)" /note="CXCR5(NM_001716)" /gene="CXCR5" CDS 2884..2904 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /transl_table=1 /translation="ENLYFQS" /label="TEV site" CDS 2905..3618 /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 3619..3642 /codon_start=1 /product="peptide that binds Strep-Tactin®, an engineered form of streptavidin" /transl_table=1 /translation="WSHPQFEK" /label="Strep-Tag II" CDS 3661..3684 /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 3756..3811 /note="rabbit β-globin polyadenylation signal (Gil and Proudfoot, 1987)" /label="β-globin poly(A) signal" primer_bind complement(4171..4187) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4195..4211 /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(4219..4249) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4264..4285 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" promoter 4344..4539 /note="SV40 early promoter" /label="SV40 promoter" polyA_signal 4545..4679 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin complement(4918..5506) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5677..6537) /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(6538..6642) /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 gatgaactac ccgctaacgc tggaaatgga cctcgagaac 1801 ctggaggacc tgttctggga actggacaga ttggacaact ataacgacac ctccctggtg 1861 gaaaatcatc tctgccctgc cacagagggg cccctcatgg cctccttcaa ggccgtgttc 1921 gtgcccgtgg cctacagcct catcttcctc ctgggcgtga tcggcaacgt cctggtgctg 1981 gtgatcctgg agcggcaccg gcagacacgc agttccacgg agaccttcct gttccacctg 2041 gccgtggccg acctcctgct ggtcttcatc ttgccctttg ccgtggccga gggctctgtg 2101 ggctgggtcc tggggacctt cctctgcaaa actgtgattg ccctgcacaa agtcaacttc 2161 tactgcagca gcctgctcct ggcctgcatc gccgtggacc gctacctggc cattgtccac 2221 gccgtccatg cctaccgcca ccgccgcctc ctctccatcc acatcacctg tgggaccatc 2281 tggctggtgg gcttcctcct tgccttgcca gagattctct tcgccaaagt cagccaaggc 2341 catcacaaca actccctgcc acgttgcacc ttctcccaag agaaccaagc agaaacgcat 2401 gcctggttca cctcccgatt cctctaccat gtggcgggat tcctgctgcc catgctggtg 2461 atgggctggt gctacgtggg ggtagtgcac aggttgcgcc aggcccagcg gcgccctcag 2521 cggcagaagg cagtcagggt ggccatcctg gtgacaagca tcttcttcct ctgctggtca 2581 ccctaccaca tcgtcatctt cctggacacc ctggcgaggc tgaaggccgt ggacaatacc 2641 tgcaagctga atggctctct ccccgtggcc atcaccatgt gtgagttcct gggcctggcc 2701 cactgctgcc tcaaccccat gctctacact ttcgccggcg tgaagttccg cagtgacctg 2761 tcgcggctcc tgacgaagct gggctgtacc ggccctgcct ccctgtgcca gctcttccct 2821 agctggcgca ggagcagtct ctctgagtca gagaatgcca cctctctcac cacgttcgaa 2881 ttcgaaaacc tgtacttcca atccgtgagc aagggcgagg agctgttcac cggggtggtg 2941 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag 3001 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag 3061 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc 3121 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac 3181 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg 3241 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag 3301 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc 3361 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag 3421 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc 3481 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac 3541 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc 3601 atggacgagc tgtacaagtg gagccacccg cagttcgaga agggcggcgg cagcagcgcc 3661 tggagccacc cgcagttcga gaagtgaaga tctttttccc tctgccaaaa attatgggga 3721 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc 3781 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca 3841 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc 3901 tgccatgaac aaaggtggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc 3961 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt 4021 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt 4081 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg aagatccctc 4141 gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 4201 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 4261 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 4321 aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta 4381 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga 4441 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag 4501 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag 4561 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 4621 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc 4681 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc 4741 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc 4801 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat 4861 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 4921 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 4981 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 5041 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 5101 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 5161 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 5221 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 5281 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 5341 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt 5401 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt 5461 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 5521 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 5581 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc 5641 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc 5701 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta 5761 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga 5821 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg 5881 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc 5941 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat 6001 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag 6061 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat 6121 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa 6181 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa 6241 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga 6301 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg 6361 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc 6421 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg 6481 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact 6541 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat 6601 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt 6661 gccacctg //