LOCUS V025074 6608 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025074 VERSION V025074 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..2817 /label="CXCR4(NM_003467)" /note="CXCR4(NM_003467)" /gene="CXCR4" CDS 2824..2844 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /transl_table=1 /translation="ENLYFQS" /label="TEV site" CDS 2845..3558 /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 3559..3582 /codon_start=1 /product="peptide that binds Strep-Tactin®, an engineered form of streptavidin" /transl_table=1 /translation="WSHPQFEK" /label="Strep-Tag II" CDS 3601..3624 /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 3696..3751 /note="rabbit β-globin polyadenylation signal (Gil and Proudfoot, 1987)" /label="β-globin poly(A) signal" primer_bind complement(4111..4127) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4135..4151 /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(4159..4189) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4204..4225 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" promoter 4284..4479 /note="SV40 early promoter" /label="SV40 promoter" polyA_signal 4485..4619 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin complement(4858..5446) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5617..6477) /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(6478..6582) /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 gatggagggg atcagtatat acacttcaga taactacacc 1801 gaggaaatgg gctcagggga ctatgactcc atgaaggaac cctgtttccg tgaagaaaat 1861 gctaatttca ataaaatctt cctgcccacc atctactcca tcatcttctt aactggcatt 1921 gtgggcaatg gattggtcat cctggtcatg ggttaccaga agaaactgag aagcatgacg 1981 gacaagtaca ggctgcacct gtcagtggcc gacctcctct ttgtcatcac gcttcccttc 2041 tgggcagttg atgccgtggc aaactggtac tttgggaact tcctatgcaa ggcagtccat 2101 gtcatctaca cagtcaacct ctacagcagt gtcctcatcc tggccttcat cagtctggac 2161 cgctacctgg ccatcgtcca cgccaccaac agtcagaggc caaggaagct gttggctgaa 2221 aaggtggtct atgttggcgt ctggatccct gccctcctgc tgactattcc cgacttcatc 2281 tttgccaacg tcagtgaggc agatgacaga tatatctgtg accgcttcta ccccaatgac 2341 ttgtgggtgg ttgtgttcca gtttcagcac atcatggttg gccttatcct gcctggtatt 2401 gtcatcctgt cctgctattg cattatcatc tccaagctgt cacactccaa gggccaccag 2461 aagcgcaagg ccctcaagac cacagtcatc ctcatcctgg ctttcttcgc ctgttggctg 2521 ccttactaca ttgggatcag catcgactcc ttcatcctcc tggaaatcat caagcaaggg 2581 tgtgagtttg agaacactgt gcacaagtgg atttccatca ccgaggccct agctttcttc 2641 cactgttgtc tgaaccccat cctctatgct ttccttggag ccaaatttaa aacctctgcc 2701 cagcacgcac tcacctctgt gagcagaggg tccagcctca agatcctctc caaaggaaag 2761 cgaggtggac attcatctgt ttccactgag tctgagtctt caagttttca ctccagcgaa 2821 ttcgaaaacc tgtacttcca atccgtgagc aagggcgagg agctgttcac cggggtggtg 2881 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag 2941 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag 3001 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc 3061 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac 3121 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg 3181 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag 3241 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc 3301 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag 3361 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc 3421 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac 3481 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc 3541 atggacgagc tgtacaagtg gagccacccg cagttcgaga agggcggcgg cagcagcgcc 3601 tggagccacc cgcagttcga gaagtgaaga tctttttccc tctgccaaaa attatgggga 3661 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc 3721 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca 3781 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc 3841 tgccatgaac aaaggtggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc 3901 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt 3961 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt 4021 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg aagatccctc 4081 gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 4141 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 4201 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 4261 aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta 4321 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga 4381 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag 4441 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag 4501 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 4561 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc 4621 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc 4681 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc 4741 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat 4801 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 4861 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 4921 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 4981 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 5041 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 5101 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 5161 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 5221 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 5281 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt 5341 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt 5401 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 5461 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 5521 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc 5581 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc 5641 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta 5701 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga 5761 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg 5821 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc 5881 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat 5941 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag 6001 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat 6061 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa 6121 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa 6181 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga 6241 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg 6301 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc 6361 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg 6421 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact 6481 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat 6541 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt 6601 gccacctg //