LOCUS Exported 5403 bp ds-DNA circular SYN 23-SEP-2022 DEFINITION This pcDNA3 plasmid contains a new poly-linker and single C-terminal flag-tag sequence for expression in mammalian cells.. ACCESSION . VERSION . KEYWORDS c-Flag pcDNA3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5403) AUTHORS Sanjabi S, Williams KJ, Saccani S, Zhou L, Hoffmann A, Ghosh G, Gerondakis S, Natoli G, Smale ST TITLE A c-Rel subdomain responsible for enhanced DNA-binding affinity and selective gene activation. JOURNAL Genes Dev. 2005 Sep 15. 19(18):2138-51. PUBMED 16166378 REFERENCE 2 (bases 1 to 5403) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 23, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5403 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(46..65) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 237..616 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 617..820 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 771..791 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 865..884 /label=T7 /note="T7 promoter, forward primer" promoter 865..883 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 922..945 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter complement(958..976) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(959..976) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(996..1013) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1002..1226 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1272..1700 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1359..1378) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1569..1590 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(1709..1729) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 1714..2043 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 1894..2029 /label=SV40 ori /note="SV40 origin of replication" primer_bind 1956..1975 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2110..2904 /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(2164..2183) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 2774..2793 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3078..3199 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3115..3134) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3169..3188 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3248..3264) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3248..3264) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3261..3283) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3272..3288 /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(3296..3326) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3341..3362 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3479..3496) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3650..4238) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3730..3749) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4409..5269) /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 5032..5051 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5270..5374) /gene="bla" /label=AmpR promoter ORIGIN 1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga 61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct 181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca 241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata 301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc 841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttatct 901 agactcgagg gtaccggatc cgactacaag gacgacgatg acaagtaaga attctattct 961 atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca 1021 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac 1081 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat 1141 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca 1201 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag 1261 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 1321 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 1381 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 1441 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 1501 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 1561 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 1621 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 1681 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc 1741 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg 1801 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 1861 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 1921 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 1981 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc 2041 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga 2101 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag 2161 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc 2221 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg 2281 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc 2341 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg 2401 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct 2461 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg 2521 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat 2581 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc 2641 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg 2701 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc 2761 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct 2821 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat 2881 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga 2941 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct 3001 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg 3061 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata 3121 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 3181 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt 3241 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 3301 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 3361 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 3421 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 3481 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 3541 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 3601 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 3661 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 3721 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 3781 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 3841 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 3901 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 3961 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 4021 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 4081 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 4141 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 4201 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 4261 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 4321 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 4381 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 4441 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 4501 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 4561 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 4621 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 4681 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 4741 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 4801 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 4861 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 4921 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 4981 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 5041 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 5101 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 5161 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 5221 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 5281 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 5341 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 5401 acg //