LOCUS Exported 5223 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Protein expression and purification of SLC9A3R1 PDZ2 domain. ACCESSION . VERSION . KEYWORDS SLC9A3R1_PDZ_2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5223) AUTHORS Tonikian R, Zhang Y, Sazinsky SL, Currell B, Yeh JH, Reva B, Held HA, Appleton BA, Evangelista M, Wu Y, Xin X, Chan AC, Seshagiri S, Lasky LA, Sander C, Boone C, Bader GD, Sidhu SS TITLE A specificity map for the PDZ domain family. JOURNAL PLoS Biol. 2008 Sep 30;6(9):e239. doi: 10.1371/journal.pbio.0060239. PUBMED 18828675 REFERENCE 2 (bases 1 to 5223) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5223 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..29 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 37..53 /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 76..729 /codon_start=1 /product="glutathione S-transferase from Schistosoma japonicum" /label=GST /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK" primer_bind 687..709 /label=pGEX 5' /note="pGEX vectors, Glutathione-S-transferase, forward primer" CDS 736..759 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" primer_bind complement(1091..1113) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(1258..1276) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1344..1448 /gene="bla" /label=AmpR promoter CDS 1449..2309 /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(1667..1686) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2480..3068 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2969..2988 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3222..3239 /label=L4440 /note="L4440 vector, forward primer" promoter 3312..3389 /gene="lacI" /label=lacI promoter CDS 3390..4472 /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 YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind complement(3409..3428) /label=LacI-R /note="LacI, reverse primer" promoter 4521..4551 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4559..4575 /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 4583..4599 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4583..4599 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 4595..5122 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR QPSEAVVWLCRS" primer_bind complement(4615..4632) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4615..4631) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4624..4646) /label=M13/pUC Forward /note="In lacZ gene" ORIGIN 1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca 61 caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa 121 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc 181 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt 241 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata 301 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt 361 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt 421 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat 481 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg 541 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa 601 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc 661 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat 721 cctccaaaat cggatctgga agttctgttc caggggcccc tgggatcccg gctctgtacc 781 atgaagaagg gccccagtgg ctatggcttc aacctgcaca gcgacaagtc caagccaggc 841 cagttcatcc ggtcagtgga cccagactcc ccggctgagg cttcagggct ccgggcccag 901 gatcgcattg tggaggtgaa cggggtctgc atggagggga agcagcatgg ggacgtggtg 961 tccgccatca gggctggcgg ggacgagacc aagctgctgg tggtggacag ggaaacttga 1021 gtcgactcga gcggccgcat cgtgactgac tgacgatctg cctcgcgcgt ttcggtgatg 1081 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg 1141 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg 1201 cagccatgac ccagtcacgt agcgatagcg gagtgtataa ttcttgaaga cgaaagggcc 1261 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag 1321 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt 1381 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa 1441 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt 1501 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt 1561 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt 1621 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg 1681 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga 1741 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa 1801 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga 1861 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa 1921 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca 1981 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta 2041 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac 2101 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc 2161 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag 2221 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga 2281 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt 2341 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata 2401 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag 2461 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 2521 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 2581 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc 2641 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 2701 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 2761 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc 2821 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa 2881 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 2941 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 3001 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 3061 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg 3121 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg 3181 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg 3241 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc 3301 gcataaattc cgacaccatc gaatggcgca aaacctttcg cggtatggca tgatagcgcc 3361 cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag 3421 agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt 3481 ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc 3541 gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc 3601 tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg 3661 gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg 3721 tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc 3781 aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct 3841 ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg 3901 tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt 3961 ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc 4021 agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc 4081 aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc 4141 tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag 4201 tgggatacga cgataccgaa gacagctcat gttatatccc gccgttaacc accatcaaac 4261 aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc 4321 aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa accacgctag 4381 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac 4441 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt aagttagctc 4501 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt 4561 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgg attcactggc 4621 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc 4681 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc 4741 ccaacagttg cgcagcctga atggcgaatg gcgctttgcc tggtttccgg caccagaagc 4801 ggtgccggaa agctggctgg agtgcgatct tcctgaggcc gatactgtcg tcgtcccctc 4861 aaactggcag atgcacggtt acgatgcgcc catctacacc aacgtgacct atcccattac 4921 ggtcaatccg ccgtttgttc ccacggagaa tccgacgggt tgttactcgc tcacatttaa 4981 tgttgatgaa agctggctac aggaaggcca gacgcgaatt atttttgatg gcgttggaat 5041 tagcttatcg actgcacggt gcaccaatgc ttctggcgtc aggcagccat cggaagctgt 5101 ggtatggctg tgcaggtcgt aaatcactgc ataattcgtg tcgctcaagg cgcactcccg 5161 ttctggataa tgttttttgc gccgacatca taacggttct ggcaaatatt ctgaaatgag 5221 ctg //