LOCUS       sequence-214708-        5510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cwc25 with C-term GST tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5510)
  TITLE     Cwc25 plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5510)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pGEX-Cwc25-GST
FEATURES             Location/Qualifiers
     source          1..5510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(48..53,54..69,70..76)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    84..100
                     /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             123..776
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     734..756
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             783..806
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1425..1447)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1592..1610)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1678..1782
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1783..1851,1852..2643)
                     /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(2001..2020)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2814..3402
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3303..3322
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3556..3573
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3646..3723
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3724..4806
                     /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(3743..3762)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4855..4860,4861..4878,4879..4885)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4893..4909
                     /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     4917..4933
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4917..4933
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4929..5456
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4949..4966)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4949..4965)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4958..4980)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgcgccgac atcataacgg ttctggcaaa tattctgaaa tgagctgttg acaattaatc
       61 atcggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag gaaacagtat
      121 tcatgtcccc tatactaggt tattggaaaa ttaagggcct tgtgcaaccc actcgacttc
      181 ttttggaata tcttgaagaa aaatatgaag agcatttgta tgagcgcgat gaaggtgata
      241 aatggcgaaa caaaaagttt gaattgggtt tggagtttcc caatcttcct tattatattg
      301 atggtgatgt taaattaaca cagtctatgg ccatcatacg ttatatagct gacaagcaca
      361 acatgttggg tggttgtcca aaagagcgtg cagagatttc aatgcttgaa ggagcggttt
      421 tggatattag atacggtgtt tcgagaattg catatagtaa agactttgaa actctcaaag
      481 ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt cgaagatcgt ttatgtcata
      541 aaacatattt aaatggtgat catgtaaccc atcctgactt catgttgtat gacgctcttg
      601 atgttgtttt atacatggac ccaatgtgcc tggatgcgtt cccaaaatta gtttgtttta
      661 aaaaacgtat tgaagctatc ccacaaattg ataagtactt gaaatccagc aagtatatag
      721 catggccttt gcagggctgg caagccacgt ttggtggtgg cgaccatcct ccaaaatcgg
      781 atctggaagt tctgttccag gggcccctgg gatccccgga attcatgggg tcgggcgatt
      841 tgaatctgtt gaaatcatgg aatcccaagc ttatgaaaaa tagaaaaaaa gtatgggaga
      901 ccgaacagga tctaattacc gaacagcaga agcttaacac gcgattgaaa gaaattgaaa
      961 aagaacgaga actaaatgaa ctgctgaacg aatcgagcaa ggataagccg gagactctca
     1021 agaatgactt ggctttgaag aagtcgggct tggaatggat gtatcaggat gcaaaactga
     1081 gtgacgaaaa ggaagactat ttattaggta agaaaaaact agattcatct atactaaacc
     1141 aacctgcaac accgcctgta agggcggcca ctactatttc cgcttctggt gccgcaacat
     1201 ccataagttc gcagaagaag aagtcaaaat tgcttaaaga tgatccaatg agtaagttta
     1261 aggtaactaa acaacaaaga agaactccag actcaacaaa aaaaagagca atgtcgcaac
     1321 gagggaaacc gttgtccaag cccgctccgg acctagacta ctgagcggcc gcatcgtgac
     1381 tgactgacga tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     1441 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     1501 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
     1561 agcggagtgt ataattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     1621 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     1681 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1741 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1801 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1861 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1921 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1981 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     2041 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     2101 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     2161 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     2221 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2281 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     2341 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2401 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2461 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2521 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     2581 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     2641 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2701 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2761 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2821 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2881 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2941 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     3001 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3061 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3121 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3181 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3241 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3301 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3361 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3421 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3481 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3541 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     3601 tttctcctta cgcatctgtg cggtatttca caccgcataa attccgacac catcgaatgg
     3661 cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
     3721 aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc
     3781 gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa
     3841 gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa
     3901 cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt
     3961 gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta
     4021 gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc
     4081 agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc
     4141 tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt
     4201 attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac
     4261 cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct
     4321 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
     4381 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
     4441 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
     4501 tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc
     4561 tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
     4621 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
     4681 cccgtctcac tggtgaaaag aaaaaccacg ctagcgccca atacgcaaac cgcctctccc
     4741 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     4801 cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcacccc aggctttaca
     4861 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     4921 aaacagctat gaccatgatt acggattcac tggccgtcgt tttacaacgt cgtgactggg
     4981 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     5041 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     5101 aatggcgctt tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg
     5161 atcttcctga ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg
     5221 cgcccatcta caccaacgtg acctatccca ttacggtcaa tccgccgttt gttcccacgg
     5281 agaatccgac gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag
     5341 gccagacgcg aattattttt gatggcgttg gaattagctt atcgactgca cggtgcacca
     5401 atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca
     5461 ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg ataatgtttt
//
