LOCUS Exported 6040 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCMV-PV-NLS-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6040) AUTHORS Pusl T, Wu JJ, Zimmerman TL, Zhang L, Ehrlich BE, Berchtold MW, Hoek JB, Karpen SJ, Nathanson MH, Bennett AM TITLE Epidermal growth factor-mediated activation of the ETS domain transcription factor Elk-1 requires nuclear calcium. JOURNAL J Biol Chem. 2002 Jul 26. 277(30):27517-27. PUBMED 11971908 REFERENCE 2 (bases 1 to 6040) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6040 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 37..413 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 415..618 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 569..589 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 675..684 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1047..1763 /codon_start=1 /product="Cycle 3 GFP (Crameri et al., 1996)" /label=Cycle 3 GFP /note="mammalian codon-optimized" /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITHGMDELYK" primer_bind complement(1080..1108) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 1703..1723 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" CDS 1776..1796 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1800..1820 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1824..1844 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1857..1886 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" primer_bind complement(1922..1939) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1928..2152 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2198..2626 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2285..2304) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2495..2516 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2635..2655) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2640..2969 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2820..2955 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2882..2901 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2995..3789 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MVEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3049..3068) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3659..3678 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3963..4084 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4000..4019) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4054..4073 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4117..4134) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4288..4876) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4368..4387) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5047..5907) /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 5670..5689 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5908..6012) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcagatcga cggatcggga gatcggtacc gaattcacat tgattattga gtagttatta 61 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata 121 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat 181 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga 241 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc 301 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt 361 atgggacttt cctacttggc agtacatcta cggttagtca tcgctattac catagtgatg 421 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt 481 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca 541 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag 601 gtctatataa gcagagcttt ctggctaact agagaaccca ctgcttactg gcacgtggaa 661 attaatacga cgtggccacc atggcccagg tgcagctgca ggtcgacatg tcgatgacag 721 acttgctcag cgctgaggac atcaagaagg cgataggagc ctttactgct gcagactcct 781 tcgaccacaa aaagttcttc cagatggtgg gcctgaagaa aaagagtgcg gatgatgtga 841 agaaggtgtt ccacattctg gacaaagaca aaagtggctt cattgaggag gatgagctgg 901 ggtccattct gaagggcttc tcctcagatg ccagagactt gtctgctaag gaaacaaaga 961 cgctgatggc tgctggagac aaggacgggg acggcaagat tggggttgaa gaattctcca 1021 ctctggtggc cgaaagcgcg gccgcaatgg ctagcaaagg agaagaactt ttcactggag 1081 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg 1141 gagagggtga aggtgatgct acatacggaa agcttaccct taaatttatt tgcactactg 1201 gaaaactacc tgttccatgg ccaacacttg tcactacttt ctcttatggt gttcaatgct 1261 tttcccgtta tccggatcat atgaaacggc atgacttttt caagagtgcc atgcccgaag 1321 gttatgtaca ggaacgcact atatctttca aagatgacgg gaactacaag acgcgtgctg 1381 aagtcaagtt tgaaggtgat acccttgtta atcgtatcga gttaaaaggt attgatttta 1441 aagaagatgg aaacattctc ggacacaaac tcgagtacaa ctataactca cacaatgtat 1501 acatcacggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt cgccacaaca 1561 ttgaagatgg atccgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg 1621 gccctgtcct tttaccagac aaccattacc tgtcgacaca atctgccctt tcgaaagatc 1681 ccaacgaaaa gcgtgaccac atggtccttc ttgagtttgt aactgctgct gggattacac 1741 atggcatgga tgagctctac aaagcggccg cagatccaaa aaagaagaga aaggtagatc 1801 caaaaaagaa gagaaaggta gatccaaaaa agaagagaaa ggtagatacg gccgcagaac 1861 aaaaactcat ctcagaagag gatctgaatg gggccgcata gtctagaagc tcgctgatca 1921 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc 1981 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg 2041 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg 2101 gaggattggg aagacaatag caggcatgct ggggatggcc cgggctctat ggcttctgag 2161 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta 2221 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg 2281 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa 2341 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctctccccc 2401 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt 2461 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca 2521 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc 2581 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg 2641 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 2701 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa 2761 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca 2821 tcccgcccct aactccgccc agttccgccc attctccgcc cctaggctga ctaatttttt 2881 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag 2941 gcttttttgg aggcctaggc ttttgcaaaa agctcccccc ccgggaggtc cacaatggtt 3001 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat 3061 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag 3121 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actccaggac 3181 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac 3241 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc 3301 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg 3361 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag 3421 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat 3481 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgtatgcc cgacggcgag 3541 gatctcgtcg tgactcatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc 3601 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg 3661 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg 3721 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag 3781 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat 3841 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 3901 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc 3961 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 4021 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 4081 cttatcatgt ctgtataccg tcgatctttc cgcttcctcg ctcactgact cgctgcgctc 4141 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 4201 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 4261 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 4321 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 4381 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 4441 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 4501 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt atgcacgaac cccccgttca 4561 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 4621 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 4681 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg 4741 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 4801 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 4861 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 4921 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 4981 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 5041 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 5101 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 5161 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 5221 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 5281 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 5341 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 5401 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 5461 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 5521 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 5581 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 5641 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 5701 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 5761 gagatccagt tcgatgtaac ccactcgggc acccaactga tcttcagcat cttttacttt 5821 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 5881 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 5941 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 6001 aggggttccg cgcacatttc cccgaaaagt gccacctgac //