LOCUS Exported 5276 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pGEX Jak2-SH2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5276) 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 5276) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5276 /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(1326..1348) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(1493..1511) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1579..1683 /gene="bla" /label=AmpR promoter CDS 1684..2544 /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(1902..1921) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2715..3303 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3204..3223 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3457..3474 /label=L4440 /note="L4440 vector, forward primer" promoter 3547..3624 /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 3625..4707 /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(3644..3663) /label=LacI-R /note="LacI, reverse primer" protein_bind 4720..4741 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4756..4786 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4794..4810 /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 4818..4834 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4818..4834 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 4830..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(4850..4867) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4850..4866) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4859..4881) /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 gaaaatatac 961 aaagcaactg tcatggccca atttcgatgg attttgccat tagtaaactg aagaaagcag 1021 gtaatcagac tggactgtat gtacttcgat gcagtcctaa ggactttaat aaatattttt 1081 tgacttttgc tgtcgagcga gaaaatgtca ttgaatataa acactgtttg attacaaaaa 1141 atgagaatga agagtacaac ctcagtggga caaagaagaa cttcagcagt cttaaagatc 1201 ttttgaattg ttaccagatg gaaactgttc gctcagacaa tataattttc cagtttacta 1261 aatgagcggc cgcatcgtga ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt 1321 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc 1381 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc 1441 atgacccagt cacgtagcga tagcggagtg tataattctt gaagacgaaa gggcctcgtg 1501 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc 1561 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat 1621 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag 1681 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt 1741 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt 1801 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc 1861 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta 1921 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac 1981 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa 2041 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg 2101 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc 2161 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg 2221 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta 2281 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg 2341 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg 2401 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc 2461 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt 2521 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt 2581 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc 2641 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag 2701 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa 2761 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg 2821 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag 2881 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg 2941 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga 3001 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc 3061 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc 3121 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga 3181 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt 3241 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg 3301 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac 3361 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga 3421 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg 3481 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata 3541 aattccgaca ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa 3601 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat 3661 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg 3721 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg 3781 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc 3841 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc 3901 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac 3961 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat 4021 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac 4081 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag 4141 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc 4201 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg 4261 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg 4321 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc 4381 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga 4441 tacgacgata ccgaagacag ctcatgttat atcccgccgt caaccaccat caaacaggat 4501 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg 4561 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc 4621 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag 4681 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca 4741 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag 4801 cggataacaa tttcacacag gaaacagcta tgaccatgat tacggattca ctggccgtcg 4861 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac 4921 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac 4981 agttgcgcag cctgaatggc gaatggcgct ttgcctggtt tccggcacca gaagcggtgc 5041 cggaaagctg gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc ccctcaaact 5101 ggcagatgca cggttacgat gcgcccatct acaccaacgt aacctatccc attacggtca 5161 atccgccgtt tgttcccacg gagaatccga cgggttgtta ctcgctcaca tttaatgttg 5221 atgaaagctg gctacaggaa ggccagacgc gaattatttt tgatggcgtt ggaatt //