LOCUS       sequence-193051-        5620 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of GST fused Nix-S212D protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5620)
  TITLE     Gordon Lab plasmids
REFERENCE   2  (bases 1 to 5620)
  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"
COMMENT     Sequence Label: pGEX4T-1-Nix-S212D
FEATURES             Location/Qualifiers
     source          1..5620
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      13..601
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     502..521
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     755..772
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        845..922
                     /gene="lacI"
                     /label=lacI promoter
     CDS             923..2005
                     /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(942..961)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2054..2059,2060..2077,2078..2084)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2092..2108
                     /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     2116..2132
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2116..2132
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2128..2655
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(2148..2165)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2148..2164)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2157..2179)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2757..2762,2763..2778,2779..2785)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2793..2809
                     /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             2832..3485
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3443..3465
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             3492..3509
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(4244..4266)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4411..4429)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4497..4601
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4602..4670,4671..5462)
                     /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(4820..4839)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
       61 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
      121 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
      181 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
      241 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
      301 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
      361 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
      421 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
      481 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
      541 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
      601 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
      661 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
      721 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
      781 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataaa
      841 ttccgacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga
      901 gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc
      961 cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa
     1021 aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc
     1081 acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct
     1141 gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag
     1201 cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa
     1261 tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc
     1321 cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca
     1381 gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca
     1441 tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc
     1501 ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat
     1561 agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct
     1621 gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc
     1681 aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata
     1741 cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca aacaggattt
     1801 tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt
     1861 gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa
     1921 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     1981 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt
     2041 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     2101 gataacaatt tcacacagga aacagctatg accatgatta cggattcact ggccgtcgtt
     2161 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     2221 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     2281 ttgcgcagcc tgaatggcga atggcgcttt gcctggtttc cggcaccaga agcggtgccg
     2341 gaaagctggc tggagtgcga tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg
     2401 cagatgcacg gttacgatgc gcccatctac accaacgtga cctatcccat tacggtcaat
     2461 ccgccgtttg ttcccacgga gaatccgacg ggttgttact cgctcacatt taatgttgat
     2521 gaaagctggc tacaggaagg ccagacgcga attatttttg atggcgttgg aattagctta
     2581 tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc tgtggtatgg
     2641 ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca aggcgcactc ccgttctgga
     2701 taatgttttt tgcgccgaca tcataacggt tctggcaaat attctgaaat gagctgttga
     2761 caattaatca tcggctcgta taatgtgtgg aattgtgagc ggataacaat ttcacacagg
     2821 aaacagtatt catgtcccct atactaggtt attggaaaat taagggcctt gtgcaaccca
     2881 ctcgacttct tttggaatat cttgaagaaa aatatgaaga gcatttgtat gagcgcgatg
     2941 aaggtgataa atggcgaaac aaaaagtttg aattgggttt ggagtttccc aatcttcctt
     3001 attatattga tggtgatgtt aaattaacac agtctatggc catcatacgt tatatagctg
     3061 acaagcacaa catgttgggt ggttgtccaa aagagcgtgc agagatttca atgcttgaag
     3121 gagcggtttt ggatattaga tacggtgttt cgagaattgc atatagtaaa gactttgaaa
     3181 ctctcaaagt tgattttctt agcaagctac ctgaaatgct gaaaatgttc gaagatcgtt
     3241 tatgtcataa aacatattta aatggtgatc atgtaaccca tcctgacttc atgttgtatg
     3301 acgctcttga tgttgtttta tacatggacc caatgtgcct ggatgcgttc ccaaaattag
     3361 tttgttttaa aaaacgtatt gaagctatcc cacaaattga taagtacttg aaatccagca
     3421 agtatatagc atggcctttg cagggctggc aagccacgtt tggtggtggc gaccatcctc
     3481 caaaatcgga tctggttccg cgtggatccc cggaattcat gtcgtcccac ctagtcgagc
     3541 cgccgccgcc cctgcacaac aacaacaaca actgcgagga aaatgagcag tctctgcccc
     3601 cgccggccgg cctcaacagt tcctgggtgg agctacccat gaacagcagc aatggcaatg
     3661 ataatggcaa tgggaaaaat ggggggctgg aacacgtacc atcctcatcc tccatccaca
     3721 atggagacat ggagaagatt cttttggatg cacaacatga atcaggacag agtagttcca
     3781 gaggcagttc tcactgtgac agcccttcgc cacaagaaga tgggcagatc atgtttgatg
     3841 tggaaatgca caccagcagg gaccatagct ctcagtcaga agaagaagtt gtagaaggag
     3901 agaaggaagt cgaggctttg aagaaaagtg cggactgggt atcagactgg tccagtagac
     3961 ccgaaaacat tccacccaag gagttccact tcagacaccc taaacgttct gtgtctttaa
     4021 gcatgaggaa aagtggagcc atgaagaaag ggggtatttt ctccgcagaa tttctgaagg
     4081 tgttcattcc atctctcttc ctttctcatg ttttggcttt ggggctaggc atctatattg
     4141 gaaagcgact ggacacaccc tctgccagca cctactgact cgagcggccg catcgtgact
     4201 gactgacgat ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     4261 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     4321 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
     4381 gcggagtgta taattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4441 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4501 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4561 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4621 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4681 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4741 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4801 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     4861 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4921 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4981 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5041 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5101 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     5161 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5221 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5281 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5341 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5401 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5461 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5521 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5581 agttttcgtt ccactgagcg tcagaccccg tagaaaagat
//
