LOCUS Exported 6211 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pGEX p85b(NC)-SH2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6211) 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 6211) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6211 /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" protein_bind 1845..1856 /label=ZF binding site /bound_moiety="CCR5TC zinc-finger domain" /note="target sequence for the CCR5TC zinc-finger domain from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 2008; Gross et al., 2013)" primer_bind complement(2261..2283) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(2428..2446) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 2514..2618 /gene="bla" /label=AmpR promoter CDS 2619..3479 /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(2837..2856) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3650..4238 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4139..4158 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4392..4409 /label=L4440 /note="L4440 vector, forward primer" promoter 4482..4559 /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 4560..5642 /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(4579..4598) /label=LacI-R /note="LacI, reverse primer" protein_bind 5655..5676 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5691..5721 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5729..5745 /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 5753..5769 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5753..5769 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 5765..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(5785..5802) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(5785..5801) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5794..5816) /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 ccggaattcg 961 gcctggccaa tgggggcagc ccaccctccc tgcaggacgc cgagtggtac tggggtgaca 1021 tctccaggga ggaggtaaac gagaaactgc gggacacgcc cgatggcacc ttcctggtcc 1081 gagatgcctc cagcaagatc cagggagagt acacgctgac cctcaggaaa ggcggaaaca 1141 acaagctgat caaggtcttc caccgagatg ggcactacgg cttctcggag ccgctcacct 1201 tctgctctgt cgtggacctc attacccact accgccatga gtcactggcc cagtacaacg 1261 ccaagctgga cacgcggctc ctctacccag tgtccaagta ccaacaggac cagatcgtca 1321 aggaggacag cgtggaggcg gtgggtgccc agctcaaggt ctaccaccag cagtaccagg 1381 acaagagccg cgagtacgac cagctctacg aggagtacac acgcacctcc caggaactgc 1441 agatgaagcg cacagccatc gaagccttca atgagaccat caagatcttt gaagagcagg 1501 gccagacgca ggagaagtgc agcaaggaat acctggagcg cttccgacgt gagggcaatg 1561 agaaagaaat gcagaggatc ctgctaaact cggagcggct caagtctcgc attgccgaga 1621 tccacgagag tcgcacaaag ctggagcagg agctgcgggc acaggcctct gacaaccgag 1681 agatcgacaa gcgcatgaac agcctcaagc cagacctcat gcagttgcgc aagatccgag 1741 accagtacct tgtgtggctc actcagaagg gtgcccggca gaagaaaatc aacgaatggt 1801 tggggatcaa gaacgagact gaggaccagt actcactgat ggaggatgag gatgacctcc 1861 cccaccacga ggaacgcacc tggtatgtag gcaagatcaa ccgcacccaa gctgaggaaa 1921 tgctgagtgg caagcgggac ggtaccttcc tcatccgtga gagcagccag cggggttgct 1981 acgcctgttc tgtggtggtg gacggggata ccaagcactg cgtcatctac cgcacggcca 2041 ccggcttcgg cttcgcggag ccctacaacc tgtacgggtc cctgaaggag ctcgtcctcc 2101 actaccagca cgcctcgctg gtgcagcaca acgacgcgct gaccgtcacg ctcgcccacc 2161 ccgtgcgcgc ccccggccct gaattcccgg gtcgactcga gcggccgcat cgtgactgac 2221 tgacgatctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc 2281 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg 2341 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg 2401 gagtgtataa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg 2461 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa 2521 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 2581 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg 2641 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc 2701 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg 2761 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga 2821 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc 2881 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag 2941 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga 3001 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg 3061 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga 3121 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt 3181 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact 3241 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt 3301 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg 3361 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta 3421 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac 3481 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta 3541 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt 3601 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt 3661 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 3721 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc 3781 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg 3841 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg 3901 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 3961 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac 4021 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg 4081 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg 4141 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat 4201 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt 4261 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg 4321 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa 4381 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc 4441 tccttacgca tctgtgcggt atttcacacc gcataaattc cgacaccatc gaatggtgca 4501 aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg 4561 tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt 4621 cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg 4681 cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt 4741 cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg 4801 cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac 4861 gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg 4921 ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca 4981 ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt 5041 tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc 5101 aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct 5161 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga 5221 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg 5281 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg 5341 ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat 5401 gttatatccc gccgtcaacc accatcaaac aggattttcg cctgctgggg caaaccagcg 5461 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg 5521 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg 5581 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt 5641 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt 5701 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac 5761 agctatgacc atgattacgg attcactggc cgtcgtttta caacgtcgtg actgggaaaa 5821 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa 5881 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg 5941 gcgctttgcc tggtttccgg caccagaagc ggtgccggaa agctggctgg agtgcgatct 6001 tcctgaggcc gatactgtcg tcgtcccctc aaactggcag atgcacggtt acgatgcgcc 6061 catctacacc aacgtaacct atcccattac ggtcaatccg ccgtttgttc ccacggagaa 6121 tccgacgggt tgttactcgc tcacatttaa tgttgatgaa agctggctac aggaaggcca 6181 gacgcgaatt atttttgatg gcgttggaat t //