LOCUS       sequence-200271-        5322 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of APBA1 PDZ1 domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5322)
  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 5322)
  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: APBA1_PDZ_1
FEATURES             Location/Qualifiers
     source          1..5322
                     /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(1190..1212)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1357..1375)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1443..1547
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1548..1616,1617..2408)
                     /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(1766..1785)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2579..3167
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3068..3087
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3321..3338
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3411..3488
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3489..4571
                     /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(3508..3527)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4620..4625,4626..4643,4644..4650)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4658..4674
                     /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     4682..4698
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4682..4698
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4694..5221
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4714..4731)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4714..4730)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4723..4745)
                     /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 tgggatctct gctcaatacc
      781 caggacatgt acaacgatga cctgatccac ttctccaagt cggaaaactg taaagatgtt
      841 ttcatagaga agcagaaagg agaaatccta ggtgtggtga ttgtggagtc tggctgggga
      901 tccatcctcc ccaccgtgat cattgccaac atgatgcatg gtggccctgc ggagaaatct
      961 gggaagctga atatcggtga ccagatcatg tccattaatg gcaccagcct ggtgggcctg
     1021 cctctgtcca cctgccagag cattattaag ggcttaaaga atcagtcccg agtcaagctg
     1081 aatatcgtga gatgtcctcc ggtgaccacc gtgttatgag tcgactcgag cggccgcatc
     1141 gtgactgact gacgatctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     1201 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     1261 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
     1321 gcgatagcgg agtgtataat tcttgaagac gaaagggcct cgtgatacgc ctatttttat
     1381 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     1441 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1501 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     1561 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1621 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     1681 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     1741 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     1801 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     1861 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     1921 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     1981 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     2041 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     2101 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     2161 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     2221 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     2281 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     2341 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     2401 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     2461 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     2521 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2581 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2641 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2701 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     2761 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2821 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2881 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2941 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3001 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3061 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3121 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3181 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3241 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3301 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     3361 ggtattttct ccttacgcat ctgtgcggta tttcacaccg cataaattcc gacaccatcg
     3421 aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg
     3481 tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc
     3541 agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag
     3601 tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg
     3661 gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc
     3721 aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga
     3781 tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac
     3841 gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag
     3901 ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca
     3961 gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg
     4021 gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc
     4081 tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag
     4141 gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg
     4201 ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta
     4261 ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag
     4321 acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc
     4381 aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc
     4441 tgttgcccgt ctcactggtg aaaagaaaaa ccacgctagc gcccaatacg caaaccgcct
     4501 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     4561 gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc accccaggct
     4621 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     4681 acaggaaaca gctatgacca tgattacgga ttcactggcc gtcgttttac aacgtcgtga
     4741 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     4801 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     4861 tggcgaatgg cgctttgcct ggtttccggc accagaagcg gtgccggaaa gctggctgga
     4921 gtgcgatctt cctgaggccg atactgtcgt cgtcccctca aactggcaga tgcacggtta
     4981 cgatgcgccc atctacacca acgtgaccta tcccattacg gtcaatccgc cgtttgttcc
     5041 cacggagaat ccgacgggtt gttactcgct cacatttaat gttgatgaaa gctggctaca
     5101 ggaaggccag acgcgaatta tttttgatgg cgttggaatt agcttatcga ctgcacggtg
     5161 caccaatgct tctggcgtca ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta
     5221 aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat gttttttgcg
     5281 ccgacatcat aacggttctg gcaaatattc tgaaatgagc tg
//
