LOCUS Exported 5882 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway destination vector. ACCESSION . VERSION . KEYWORDS pGCS-C2m(3xHA) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5882) AUTHORS Wang HY, Li Y, Xue T, Cheng N, Du HN TITLE Construction of a series of pCS2+ backbone-based Gateway vectors for overexpressing various tagged proteins in vertebrates. JOURNAL Acta Biochim Biophys Sin (Shanghai). 2016 Oct 26. PUBMED 27797719 REFERENCE 2 (bases 1 to 5882) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5882 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 35..53 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 35..52 /label=SP6 /note="SP6 promoter, forward primer" protein_bind 93..217 /gene="mutant version of attR" /label=attR1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 254..284 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" CDS 338..997 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(404..423) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" CDS 1339..1644 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 1523..1542 /label=ccdB-fwd /note="ccdB gene, forward primer" protein_bind complement(1685..1809) /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 1826..1906 /codon_start=1 /product="three tandem HA epitope tags" /label=3xHA /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA" promoter complement(1912..1929) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 1934..2068 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1958..1977) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2012..2031 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2181..2201) /label=T3 /note="T3 promoter, forward primer" promoter complement(2181..2199) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2220..2236) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2220..2236) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2233..2255) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2244..2260 /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(2268..2298) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2313..2334 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2451..2468) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2622..3210) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2702..2721) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3381..4241) /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 4004..4023 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4242..4346) /gene="bla" /label=AmpR promoter rep_origin complement(4372..4827) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4509..4530) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4721..4740 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4905..5882 /label=CMV IE94 promoter /note="enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94" ORIGIN 1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag 61 ctacttgttc tttttgcagg atccggccgg ccacaagttt gtacaaaaaa gctgaacgag 121 aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta 181 cataatactg taaaacacaa catatccagt cactatggcg gcccctcgag gggccgcatt 241 aggcacccca ggctttacac tttatgcttc cggctcgtat aatgtgtgga ttttgagtta 301 ggatccgtcg agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata 361 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt 421 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt 481 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa 541 tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt 601 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga 661 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg 721 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa 781 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc 841 ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc 901 gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt 961 acaacagtac tgcgatgagt ggcagggcgg ggcgtaaacg cgtggatccg gcttactaaa 1021 agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa tatatactga 1081 tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat tacagtgaca 1141 gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc tccggtctgg 1201 taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa 1261 atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct tttgctgacg 1321 agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga gagccgttat 1381 cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg gatggtgatc 1441 cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg 1501 catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc 1561 gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt 1621 aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca gtctgcaggt 1681 cgaccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa 1741 aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag ctttcttgta 1801 caaagtggtg aagacaggat ccatgtaccc atacgatgtt ccagattacg cttatcctta 1861 tgacgtacct gactatgcat acccttatga tgtaccagac tacgcttaga actatagtga 1921 gtcgtattac gtagatccag acatgataag atacattgat gagtttggac aaaccacaac 1981 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt 2041 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca 2101 ggttcagggg gaggtgtggg aggtttttta attcgcggcc gcggcgccaa tgcattgggc 2161 ccggtaccca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg 2221 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 2281 gggagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 2341 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 2401 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 2461 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 2521 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 2581 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 2641 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 2701 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 2761 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 2821 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 2881 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 2941 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 3001 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 3061 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 3121 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 3181 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 3241 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 3301 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 3361 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 3421 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 3481 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 3541 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 3601 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 3661 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 3721 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 3781 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 3841 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 3901 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 3961 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 4021 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 4081 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 4141 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 4201 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 4261 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 4321 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa 4381 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc 4441 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga 4501 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag 4561 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt 4621 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta 4681 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag 4741 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg 4801 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg 4861 aagggcgatc ggtgcgggcc tcttcgctat tacgccagtc gaccgccaat tcaatatggc 4921 gtatatggac tcatgccaat tcaatatggt ggatctggac ctgtgccaat tcaatatggc 4981 gtatatggac tcgtgccaat tcaatatggt ggatctggac cccagccaat tcaatatggc 5041 ggacttggca ccatgccaat tcaatatggc ggacctggca ctgtgccaac tggggagggg 5101 tctacttggc acggtgccaa gtttgaggag gggtcttggc cctgtgccaa gtccgccata 5161 ttgaattggc atggtgccaa taatggcggc catattggct atatgccagg atcaatatat 5221 aggcaatatc caatatggcc ctatgccaat atggctattg gccaggttca atactatgta 5281 ttggccctat gccatatagt attccatata tgggttttcc tattgacgta gatagcccct 5341 cccaatgggc ggtcccatat accatatatg gggcttccta ataccgccca tagccactcc 5401 cccattgacg tcaatggtct ctatatatgg tctttcctat tgacgtcata tgggcggtcc 5461 tattgacgta tatggcgcct cccccattga cgtcaattac ggtaaatggc ccgcctggct 5521 caatgcccat tgacgtcaat aggaccaccc accattgacg tcaatgggat ggctcattgc 5581 ccattcatat ccgttctcac gccccctatt gacgtcaatg acggtaaatg gcccacttgg 5641 cagtacatca atatctatta atagtaactt ggcaagtaca ttactattgg aagtacgcca 5701 gggtacattg gcagtactcc cattgacgtc aatggcggta aatggcccgc gatggctgcc 5761 aagtacatcc ccattgacgt caatggggag gggcaatgac gcaaatgggc gttccattga 5821 cgtaaatggg cggtaggcgt gcctaatggg aggtctatat aagcaatgct cgtttaggga 5881 ac //