LOCUS Exported 5333 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pGEX Stat3-SH2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5333) AUTHORS Machida K, Thompson CM, Dierck K, Jablonowski K, Karkkainen S, Liu B, Zhang H, Nash PD, Newman DK, Nollau P, Pawson T, Renkema GH, Saksela K, Schiller MR, Shin DG, Mayer BJ TITLE High-throughput phosphotyrosine profiling using SH2 domains. JOURNAL Mol Cell. 2007 Jun 22;26(6):899-915. PUBMED 17588523 REFERENCE 2 (bases 1 to 5333) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5333 /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(1383..1405) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(1550..1568) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1636..1740 /gene="bla" /label=AmpR promoter CDS 1741..2601 /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(1959..1978) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2772..3360 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3261..3280 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3514..3531 /label=L4440 /note="L4440 vector, forward primer" promoter 3604..3681 /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 3682..4764 /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(3701..3720) /label=LacI-R /note="LacI, reverse primer" protein_bind 4777..4798 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4813..4843 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4851..4867 /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 4875..4891 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4875..4891 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 4887..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(4907..4924) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4907..4923) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4916..4938) /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 tacatcatgg 961 gctttatcag taaggagcgg gagcgggcca tcttgagcac taagcctcca ggcaccttcc 1021 tgctaagatt cagtgaaagc agcaaagaag gaggcgtcac tttcacttgg gtggagaagg 1081 acatcagcgg taagacccag atccagtccg tggaaccata cacaaagcag cagctgaaca 1141 acatgtcatt tgctgaaatc atcatgggct ataagatcat ggatgctacc aatatcctgg 1201 tgtctccact ggtctatctc tatcctgaca ttcccaagga ggaggcattc ggaaagtatt 1261 gtcggccaga gagccaggag catcctgaag ctgacccagg tggaattccc gggtcgactc 1321 gagcggccgc atcgtgactg actgacgatc tgcctcgcgc gtttcggtga tgacggtgaa 1381 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg 1441 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg 1501 acccagtcac gtagcgatag cggagtgtat aattcttgaa gacgaaaggg cctcgtgata 1561 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact 1621 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg 1681 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt 1741 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct 1801 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca 1861 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc 1921 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc 1981 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg 2041 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta 2101 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc 2161 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt 2221 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg 2281 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct 2341 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc 2401 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct 2461 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac 2521 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc 2581 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat 2641 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 2701 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 2761 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 2821 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 2881 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta 2941 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 3001 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 3061 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 3121 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 3181 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 3241 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 3301 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 3361 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 3421 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct 3481 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa 3541 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataaat 3601 tccgacacca tcgaatggtg caaaaccttt cgcggtatgg catgatagcg cccggaagag 3661 agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc 3721 ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa 3781 acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca 3841 caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg 3901 cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc 3961 gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat 4021 cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc 4081 attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag 4141 acacccatca acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat 4201 ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg 4261 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata 4321 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg 4381 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca 4441 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac 4501 gacgataccg aagacagctc atgttatatc ccgccgtcaa ccaccatcaa acaggatttt 4561 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg 4621 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat 4681 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt 4741 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta 4801 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg 4861 ataacaattt cacacaggaa acagctatga ccatgattac ggattcactg gccgtcgttt 4921 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc 4981 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt 5041 tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa gcggtgccgg 5101 aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc tcaaactggc 5161 agatgcacgg ttacgatgcg cccatctaca ccaacgtaac ctatcccatt acggtcaatc 5221 cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt aatgttgatg 5281 aaagctggct acaggaaggc cagacgcgaa ttatttttga tggcgttgga att //