LOCUS Exported 5505 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION pcDNA3 vector with added Flag, HA tags. ACCESSION . VERSION . KEYWORDS 1436 pcDNA3 Flag HA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5505) TITLE Vectors REFERENCE 2 (bases 1 to 5505) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5505 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(32..51) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 223..602 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 603..806 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 757..777 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 851..870 /label=T7 /note="T7 promoter, forward primer" promoter 851..869 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 889..912 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 925..951 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" promoter complement(1046..1064) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(1047..1064) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(1084..1101) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1090..1314 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1360..1788 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1447..1466) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1657..1678 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(1797..1817) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 1802..2131 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 1982..2117 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2044..2063 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2198..2992 /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(2252..2271) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 2862..2881 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3166..3287 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3203..3222) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3257..3276 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3336..3352) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3336..3352) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3349..3371) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3360..3376 /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(3384..3414) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3429..3450 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3567..3584) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3738..4326) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3818..3837) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4497..5357) /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 5120..5139 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5358..5462) /gene="bla" /label=AmpR promoter ORIGIN 1 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa 61 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt 121 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc 181 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta 241 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 301 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 361 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 421 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 481 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 541 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 601 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat 661 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 721 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac 781 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc 841 ttatcgaaat taatacgact cactataggg agacccaagc ttaccatgga ctacaaggac 901 gacgatgaca agctcgatgg aggatacccc tacgacgtgc ccgactacgc cagcggatcc 961 actagtaacg gccgccagtg tgctggaatt ctgcagatat ccatcacact ggcggccgct 1021 cgagcatgca tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat 1081 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt 1141 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat 1201 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg 1261 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg 1321 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat 1381 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag 1441 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc 1501 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc 1561 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt 1621 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa 1681 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg 1741 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa 1801 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 1861 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag 1921 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc 1981 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt 2041 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg 2101 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt 2161 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca 2221 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac 2281 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt 2341 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc 2401 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg 2461 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc 2521 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc 2581 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat 2641 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc 2701 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca 2761 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga 2821 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat 2881 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc 2941 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact 3001 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc 3061 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg 3121 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca 3181 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 3241 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata 3301 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat 3361 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg 3421 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag 3481 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt 3541 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 3601 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 3661 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 3721 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 3781 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 3841 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 3901 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 3961 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 4021 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 4081 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 4141 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 4201 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 4261 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 4321 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 4381 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 4441 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 4501 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 4561 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 4621 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag 4681 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 4741 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 4801 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 4861 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 4921 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 4981 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 5041 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 5101 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 5161 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 5221 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 5281 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 5341 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 5401 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 5461 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cggga //