LOCUS Exported 5359 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pGEX p85b(C)-SH2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5359) 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 5359) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5359 /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(1409..1431) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(1576..1594) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1662..1766 /gene="bla" /label=AmpR promoter CDS 1767..2627 /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(1985..2004) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2798..3386 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3287..3306 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3540..3557 /label=L4440 /note="L4440 vector, forward primer" promoter 3630..3707 /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 3708..4790 /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(3727..3746) /label=LacI-R /note="LacI, reverse primer" protein_bind 4803..4824 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4839..4869 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4877..4893 /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 4901..4917 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4901..4917 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 4913..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(4933..4950) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4933..4949) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4942..4964) /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 gggcgcgcca 961 tggacgagga cgatctcccg caccacgagg aacgcacttg gtacgtgggc aagatcaacc 1021 gcacgcaggc agaggagatg ctgagtggca agcgggatgg caccttcctc atccgcgaga 1081 gcagccagcg gggctgctac gcctgctccg tggtagtgga cggcgacacc aagcactgcg 1141 tcatctaccg cacggccacc ggcttcggct tcgcggagcc ctacaacctg tacgggtcgc 1201 tgaaggagct ggtgctgcac taccagcacg cctcgctggt gcagcacaac gacgcgctca 1261 ccgtcaccct ggcgcaccca gtgcgcgccc cgggccccgg cccgccgcct gccgcccgct 1321 taattaacga attcccgggt cgactcgagc ggccgcatcg tgactgactg acgatctgcc 1381 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 1441 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 1501 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtataatt 1561 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa 1621 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt 1681 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc 1741 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc 1801 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa 1861 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg 1921 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag 1981 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc 2041 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta 2101 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg 2161 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca 2221 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac 2281 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat 2341 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg 2401 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata 2461 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta 2521 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa 2581 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag 2641 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg 2701 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact 2761 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 2821 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 2881 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 2941 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 3001 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 3061 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 3121 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 3181 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 3241 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 3301 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 3361 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 3421 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata 3481 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca 3541 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc 3601 tgtgcggtat ttcacaccgc ataaattccg acaccatcga atggtgcaaa acctttcgcg 3661 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa 3721 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga 3781 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc 3841 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg 3901 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat 3961 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg 4021 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta 4081 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg 4141 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag 4201 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt 4261 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc 4321 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg 4381 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg 4441 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg 4501 gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc 4561 cgtcaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc 4621 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga 4681 aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt 4741 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca 4801 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct 4861 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat 4921 gattacggat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac 4981 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc 5041 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gctttgcctg 5101 gtttccggca ccagaagcgg tgccggaaag ctggctggag tgcgatcttc ctgaggccga 5161 tactgtcgtc gtcccctcaa actggcagat gcacggttac gatgcgccca tctacaccaa 5221 cgtaacctat cccattacgg tcaatccgcc gtttgttccc acggagaatc cgacgggttg 5281 ttactcgctc acatttaatg ttgatgaaag ctggctacag gaaggccaga cgcgaattat 5341 ttttgatggc gttggaatt //