LOCUS Exported 4119 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION CMV and T7 promoter plasmid for EGFP expression in mammalian cells or for in vitro transcription. ACCESSION . VERSION . KEYWORDS pCMV-T7-EGFP (BPK1098) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4119) TITLE CMV and T7 promoter plasmid to express EGFP (BPK1098) JOURNAL Unpublished REFERENCE 2 (bases 1 to 4119) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4119 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 132..335 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 286..306 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 377..396 /label=T7 /note="T7 promoter, forward primer" promoter 377..395 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 403..412 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 409..1125 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(454..475) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(715..734) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1062..1083 /label=EGFP-C /note="EGFP, forward primer" CDS 1134..1151 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(1174..1191) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1180..1404 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(1475..1491) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1475..1491) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1488..1510) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1499..1515 /label=lac operator /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-beta-D-thiogalactopyranoside (IPTG)." promoter complement(1523..1553) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1568..1589 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1706..1723) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1877..2465) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1957..1976) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2636..3496) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 3259..3278 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3497..3601) /gene="bla" /label=AmpR promoter primer_bind complement(3680..3699) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 3871..131 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg 61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg 121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact 181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa 241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta 301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct 361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat ggtgagcaag 421 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac 481 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc 541 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc 601 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc 661 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac 721 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc 781 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac 841 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg 901 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag 961 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc 1021 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc 1081 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtgacc ggtcatcatc 1141 accatcacca ttgagtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc 1201 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg 1261 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc 1321 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg 1381 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctcgatac 1441 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt 1501 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctagg 1561 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt 1621 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt 1681 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc 1741 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg 1801 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg 1861 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac 1921 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 1981 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 2041 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg 2101 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct 2161 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac 2221 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt 2281 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc 2341 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca 2401 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 2461 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 2521 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 2581 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc 2641 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 2701 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg 2761 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 2821 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta 2881 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 2941 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 3001 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 3061 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg 3121 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga 3181 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 3241 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 3301 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt 3361 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 3421 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 3481 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt 3541 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc 3601 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatcga tctcccgatc 3661 ccctagggtc gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc 3721 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa 3781 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc 3841 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt 3901 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta 3961 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccaatgacg tcaataatga 4021 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt 4081 tacggtaaac tgcccacttg gcagtacatc aagtgtatc //