LOCUS       sequence-199756-        5319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of Multiple PDZ protein PDZ12 
            domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5319)
  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 5319)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS 
            Biol."; date: "2008-09-30"; volume: "6(9):e239. doi"; pages: " 
            10.1371/journal.pbio.0060239"
COMMENT     Sequence Label: MPDZ_PDZ_12
FEATURES             Location/Qualifiers
     source          1..5319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..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(1187..1209)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1354..1372)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1440..1544
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1545..1613,1614..2405)
                     /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(1763..1782)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2576..3164
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3065..3084
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3318..3335
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3408..3485
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3486..4568
                     /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(3505..3524)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4617..4622,4623..4640,4641..4647)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4655..4671
                     /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     4679..4695
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4679..4695
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4691..5218
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4711..4728)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4711..4727)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4720..4742)
                     /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 tgggatccag ctcaaagaag
      781 aatgcattgg catctgaaat acagggatta agaacagtcg aaatgaaaaa gggccctact
      841 gactcactgg gaatcagcat cgctggagga gtaggcagcc cacttggtga tgtgcctata
      901 tttattgcaa tgatgcaccc aactggagtt gcagcacaga cccaaaaact cagagttggg
      961 gataggattg tcaccatctg tggcacatcc actgagggca tgactcacac ccaagcagtt
     1021 aacctactga aaaatgcatc tggctccatt gaaatgcagg tggttgctgg aggagacgtg
     1081 agtgtggtca caggtcatca gcaggagcct gcatgagtcg actcgagcgg ccgcatcgtg
     1141 actgactgac gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     1201 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     1261 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
     1321 atagcggagt gtataattct tgaagacgaa agggcctcgt gatacgccta tttttatagg
     1381 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     1441 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1501 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     1561 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     1621 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     1681 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     1741 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
     1801 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     1861 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     1921 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     1981 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2041 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     2101 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2161 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2221 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2281 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2341 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2401 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     2461 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     2521 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2581 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2641 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2701 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     2761 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2821 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2881 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2941 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3001 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3061 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3121 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3181 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3241 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3301 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     3361 attttctcct tacgcatctg tgcggtattt cacaccgcat aaattccgac accatcgaat
     3421 ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg
     3481 tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga
     3541 ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg
     3601 aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca
     3661 aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa
     3721 ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg
     3781 tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg
     3841 tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg
     3901 cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta
     3961 ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc
     4021 accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg
     4081 ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg
     4141 actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc
     4201 ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg
     4261 agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca
     4321 gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa
     4381 ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt
     4441 tgcccgtctc actggtgaaa agaaaaacca cgctagcgcc caatacgcaa accgcctctc
     4501 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     4561 ggcagtgagc gcaacgcaat taatgtaagt tagctcactc attaggcacc ccaggcttta
     4621 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     4681 ggaaacagct atgaccatga ttacggattc actggccgtc gttttacaac gtcgtgactg
     4741 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     4801 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     4861 cgaatggcgc tttgcctggt ttccggcacc agaagcggtg ccggaaagct ggctggagtg
     4921 cgatcttcct gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc acggttacga
     4981 tgcgcccatc tacaccaacg tgacctatcc cattacggtc aatccgccgt ttgttcccac
     5041 ggagaatccg acgggttgtt actcgctcac atttaatgtt gatgaaagct ggctacagga
     5101 aggccagacg cgaattattt ttgatggcgt tggaattagc ttatcgactg cacggtgcac
     5161 caatgcttct ggcgtcaggc agccatcgga agctgtggta tggctgtgca ggtcgtaaat
     5221 cactgcataa ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg
     5281 acatcataac ggttctggca aatattctga aatgagctg
//
