LOCUS Exported 5710 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial expression of SF2. ACCESSION . VERSION . KEYWORDS pGEX6P-1-SF2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5710) AUTHORS Huang X, Zheng M, Wang P, Mok BW, Liu S, Lau SY, Chen P, Liu YC, Liu H, Chen Y, Song W, Yuen KY, Chen H TITLE An NS-segment exonic splicing enhancer regulates influenza A virus replication in mammalian cells. JOURNAL Nat Commun. 2017 Mar 21;8:14751. doi: 10.1038/ncomms14751. PUBMED 28323816 REFERENCE 2 (bases 1 to 5710) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5710 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 183..211 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 219..235 /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)." CDS 258..911 /codon_start=1 /product="glutathione S-transferase from Schistosoma japonicum" /label=GST /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK" primer_bind 869..891 /label=pGEX 5' /note="pGEX vectors, Glutathione-S-transferase, forward primer" CDS 918..941 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" primer_bind complement(1760..1782) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(1927..1945) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 2013..2117 /gene="bla" /label=AmpR promoter CDS 2118..2978 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(2336..2355) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3149..3737 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3638..3657 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3891..3908 /label=L4440 /note="L4440 vector, forward primer" promoter 3981..4058 /gene="lacI (mutant)" /label=lacIq promoter /note="In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold." CDS 4059..5141 /codon_start=1 /gene="lacI" /product="lac repressor" /label=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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind complement(4078..4097) /label=LacI-R /note="LacI, reverse primer" protein_bind 5154..5175 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5190..5220 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5228..5244 /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)." primer_bind 5252..5268 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5252..5268 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 5264..81 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGITLSTARCTNASGVR QPSEAVVWLCRS" primer_bind complement(5284..5301) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(5284..5300) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5293..5315) /label=M13/pUC Forward /note="In lacZ gene" ORIGIN 1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg 61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt 121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc 181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca 241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc 301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc 361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc 421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata 481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc 541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact 601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag 661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt 721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa 781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat 841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc 901 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc tcgggaggtg 961 gtgtgattcg tggccccgca gggaacaacg attgccgcat ctacgtgggt aacttacctc 1021 cagacatccg aaccaaggac attgaggacg tgttctacaa atacggcgct atccgcgaca 1081 tcgacctcaa gaatcgccgc gggggaccgc ccttcgcctt cgttgagttc gaggacccgc 1141 gagacgcgga agacgcggtg tatggtcgcg acggctatga ttacgatggg taccgtctgc 1201 gggtggagtt tcctcgaagc ggccgtggaa caggccgagg cggcggcggg ggtggaggtg 1261 gcggagctcc ccgaggtcgc tatggccccc catccaggcg gtctgaaaac agagtggttg 1321 tctctggact gcctccaagt ggaagttggc aggatttaaa ggatcacatg cgtgaagcag 1381 gtgatgtatg ttatgctgat gtttaccgag atggcactgg tgtcgtggag tttgtacgga 1441 aagaagatat gacctatgca gttcgaaaac tggataacac taagtttaga tctcatgagg 1501 gagaaactgc ctacatccgg gttaaagttg atgggcccag aagtccaagt tatggaagat 1561 ctcgatctcg aagccgtagt cgtagcagaa gccgtagcag aagcaacagc aggagtcgca 1621 gttactcccc aaggagaagc agaggatcac cacgctattc tccccgtcat agcagatctc 1681 gctctcgtac ataactcgag cggccgcatc gtgactgact gacgatctgc ctcgcgcgtt 1741 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc 1801 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt 1861 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtataat tcttgaagac 1921 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt 1981 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct 2041 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat 2101 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg 2161 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg 2221 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc 2281 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat 2341 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact 2401 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca 2461 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact 2521 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg 2581 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg 2641 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg 2701 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg 2761 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag 2821 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc 2881 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga 2941 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat 3001 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc 3061 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag 3121 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct 3181 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac 3241 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc 3301 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg 3361 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt 3421 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt 3481 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc 3541 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca 3601 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata 3661 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg 3721 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct 3781 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta 3841 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag 3901 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta 3961 tttcacaccg cataaattcc gacaccatcg aatggtgcaa aacctttcgc ggtatggcat 4021 gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg 4081 atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca 4141 gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca 4201 ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca 4261 cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg 4321 atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta 4381 aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc 4441 tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc 4501 ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc 4561 gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc 4621 cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca 4681 atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac 4741 aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc 4801 agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata 4861 tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg ccgtcaacca 4921 ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct 4981 ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa 5041 ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc 5101 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg 5161 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg 5221 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgga 5281 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa 5341 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga 5401 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc 5461 accagaagcg gtgccggaaa gctggctgga gtgcgatctt cctgaggccg atactgtcgt 5521 cgtcccctca aactggcaga tgcacggtta cgatgcgccc atctacacca acgtaaccta 5581 tcccattacg gtcaatccgc cgtttgttcc cacggagaat ccgacgggtt gttactcgct 5641 cacatttaat gttgatgaaa gctggctaca ggaaggccag acgcgaatta tttttgatgg 5701 cgttggaatt //