LOCUS       sequence-145562-        8839 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GST fusion Mustela putorius furo Sacsin 94-1381.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8839)
  TITLE     Sacsin | SIRPT1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8839)
  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            "Unpublished"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-SIRPT1-Mustela putorius furo
FEATURES             Location/Qualifiers
     source          1..8839
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..941
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(4889..4911)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5056..5074)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5142..5246
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5247..5315,5316..6107)
                     /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(5465..5484)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6278..6866
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6767..6786
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7020..7037
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        7110..7187
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             7188..8270
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(7207..7226)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    8283..8304
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8319..8324,8325..8342,8343..8349)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8357..8373
                     /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     8381..8397
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8381..8397
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(8393..8839,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGITLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(8413..8430)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8413..8429)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8422..8444)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc catatgtttg
      961 gccaaacgac tcctcctctg gtagacttcc tgaaggacat cctgcgtcgt taccctgaag
     1021 gtggtcaaat cctgaaggaa ctgatccaaa acgcagaaga tgcaggtgca actgaagtca
     1081 agttcctgta cgacgagact caatacggta ccgagactct gtggtctaag gagatggcac
     1141 aataccaggg tccagcactg tacgtttaca acaacgctgt gttcacccct gaggactggc
     1201 atggtatcca agagatcgct cgttctcgta agaaggacga ccctctgaaa gtaggtcgtt
     1261 tcggtatcgg tttcaactcc gtataccaca tcaccgacgt cccttgtatc ttctctggtg
     1321 accaaatcgg tatgctggac cctcaccaaa ctctgttcgg tccacatgag tctggtcagt
     1381 gttggaacct gaaggacgat tctaaggaga tctccgagct gtccgatcag tttgcaccat
     1441 tcatcggtat cttcggttcc accaaggaga ctttcgtcaa cgctaacttc ccgggtactt
     1501 tcttccgctt cccactgcgt ctgcagccat ctcagctgtc ttctaacctg tacaacaagc
     1561 agaaggtcct ggaactgttc gactctttcc gtgcagacgc agatactgta ctgctgttcc
     1621 tgaagtctgt gcaggacgtt tctctgcacg tacgtgaggc agatggtact aagcgtctgg
     1681 ttttccgtgt aactgcatct gagaacaagg cactgaagca cgagcgtcca aactctgtaa
     1741 agatcctgga gacggcaatc tctaactact gcaagaagat cccgtccaac tccatcacgt
     1801 gcgtgaccta ccacatcaac atcgtcctgg aggacgagtc tactaaggac gctcagaaga
     1861 cttcttggct ggtatgtaac tccgtgggtg gtcgtggtat ttctggtaag ctggactctc
     1921 tggcagatga gctgaaattc gtcccaatca tcggtatcgc tatgagcctg tctcgtgacg
     1981 acgaagagaa aggtgctacc gcagatttct ctggtaaagc tttctgcttc ctgccactgc
     2041 caccaggtga agagtctaaa accggtctgc caatccacat ctctggtttc ttcggtctga
     2101 ctgacaaccg tcgttctatc aaatggcgcg agctggacca gtggcgtgat ccggcagctc
     2161 tgtggaatga atttctggta gtaaacgtcg ttccgaaagc ttacgcaact ctgatcctgg
     2221 actctatcaa acgcctggaa accgaacgtt cttccgactt cccgctgtct gtagacatca
     2281 tctacaaact gtggccggac gcaaacaaag tgaaagtcca ctggcagccg gttctggaac
     2341 cgctgtttaa cgaactgttc cagaacgctg tcctgtactc tatctccaac cactgggtca
     2401 aactggaaga agcttacttc tccgaactgg acgaatccct ggaatacacg aaaaccgtgc
     2461 tgtcctacct gcagagcagc ggtaaacaga tcgtcaaagt gccggctaac ctggctgccg
     2521 ccgtacagct ggctgcttct tctgctaaac cggtacgtaa agtaactccg gcttgggtac
     2581 gtcaggttct gcgtaaatct gttcagccgg gtgatgctca ggataaactg catctgctgg
     2641 aattcgttct gtctgaccag gcttattctg aactgctggg cctggaactg ctgcctctgc
     2701 agaatggcaa ttttgtaccg ttttctagca gcgtgagcga ccaggatgtt atctacatca
     2761 ccagcgaaga ctacccgcgt tctctgttcc cgggcctgga aggccgtttt attctggata
     2821 acctgaaacc gcatctgatc gcggctctga aagaagccgc gcagactcgt gcccaacagc
     2881 tgctgtgggg ctattctggc cgtccgtgta ctcaactgca gctgctgaat ccggaacgtt
     2941 ttgcgcgtct gattaaagaa gttatgaata ccttttggcc gggccgtgaa ctgattgttc
     3001 agtggtatcc gtttgatgaa gataaaaatc atccgtctgt tgcctggctg aaaatggttt
     3061 ggaaaaacct gtacatccac ttcagcgaag acctgacgct gttcgacgaa atgccgctga
     3121 tcccgcgtac gaccctggaa gaaggccaga aatgtgttga actgatccgc ctgcgcattc
     3181 cgtctctggt tattctggat gatgaatctg aagcgcagct gccggaattt ctggccgata
     3241 ttgttcagaa actgggcggc attgttctga aaaaactgga cgcctccatc cagcatccgc
     3301 tgattaaaaa atacatccac agcccgctgc cgtccgcggt tctgcagatt atggaaaaaa
     3361 tgccgctgca gaaactgtgt aaccagatcg cgtccctgct gccgacgcat aaagatgcgc
     3421 tgcgcaaatt tctggcgtct ctgaccgata tttccgaaaa agaaaaacgc atcatccagg
     3481 aactgccgat cttcaaacgc atcaaccacc tgtccgacca gggcctgagc agctatacca
     3541 aactgaaagg ctgtaaagtg ctgcaccaca ccgcgaaact gctgccggat ctgcgcctga
     3601 gcatttccgt tattgatagc agcgatgaag ccaccattcg cctggcgaat atgctgaaaa
     3661 ttgaaaaact gaaaaccacc aactgcctga aactgatcct gaaagacatc gaaaacgcgt
     3721 tctacagcca cgacgaagtg acccacctga tgctgtggac cctggaaaac ctgtcctccc
     3781 tgaaaaacga aaacccgaac gtgctggact ggctgatgcc gctgaaattc atccagatca
     3841 gccaggaaca gatggtgagc gcgggcgaac tgtttgatcc ggatattgaa gttctgaaag
     3901 acctgttcta cagcgaagag gagacctgct tcccgccgtc cgttttcacc agcccggata
     3961 ttctgcattc cctgcgccaa attggcctga aaaatgaagc cagcctgaaa gaaaacgacg
     4021 tggtgcaggt tgcgaaaaac atcgaagccc tgcaggttag ctcctgcccg aatcaggatg
     4081 ttctgctgaa aaaagcgaaa acgctgctgc tggttctgaa caaaaaccac accctgctgc
     4141 agagctccga aggcaaaatg accctgaaaa aaatcaaatg ggtgccggcg tgcaaagaac
     4201 gcccgccgaa ctatccgggc agcctggttt ggaaaggcga tgtttgcaac ctgtgcgcgc
     4261 cgccggatat gtgcgatggc gcccatgcga ttctggttgg cagctccctg ccgctggttg
     4321 aatccgttca tattaacctg gaaaaagcgc tgggcatctt taccaaaccg tccatcagcg
     4381 cggttctgaa acactttaaa atcgttgtgg attggtatac ctccaaaacc tttagcgatg
     4441 aagattatta tcagttccag cacatcctgc tggaaattta tggcttcatg cacgaccacc
     4501 tgaacgaagg caaagacgcg ttccgcgccc tgaaattccc gtgggtgtgg accggcaaaa
     4561 aattttgccc gctggcgcag gcggtgatta aaccgtttca cgatctggat ctgcagccgt
     4621 atctgcactc cgtgccgaaa accatggcga aattccacca gctgttcaaa gtgtgcggca
     4681 gcattgaaga actgacctcc gaccacatta gcatggtgat tcagaaagtg tatctgaaat
     4741 ccgaccagga cctgagcgat caggaatcca aacagaacct gcatctgatg ctgaacatta
     4801 ttcgctggct gtattccaac taactcgagc ggccgcatcg tgactgactg acgatctgcc
     4861 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     4921 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     4981 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtataatt
     5041 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     5101 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5161 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5221 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     5281 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5341 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     5401 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     5461 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc
     5521 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     5581 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5641 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     5701 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5761 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     5821 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5881 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5941 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6001 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6061 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6121 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6181 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6241 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6301 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6361 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6421 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6481 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6541 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6601 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6661 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6721 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6781 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6841 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6901 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6961 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     7021 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     7081 tgtgcggtat ttcacaccgc ataaattccg acaccatcga atggtgcaaa acctttcgcg
     7141 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
     7201 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     7261 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
     7321 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
     7381 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
     7441 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
     7501 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
     7561 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg
     7621 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag
     7681 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
     7741 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc
     7801 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg
     7861 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg
     7921 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg
     7981 gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc
     8041 cgtcaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
     8101 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga
     8161 aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     8221 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     8281 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     8341 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     8401 gattacggat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     8461 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     8521 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gctttgcctg
     8581 gtttccggca ccagaagcgg tgccggaaag ctggctggag tgcgatcttc ctgaggccga
     8641 tactgtcgtc gtcccctcaa actggcagat gcacggttac gatgcgccca tctacaccaa
     8701 cgtaacctat cccattacgg tcaatccgcc gtttgttccc acggagaatc cgacgggttg
     8761 ttactcgctc acatttaatg ttgatgaaag ctggctacag gaaggccaga cgcgaattat
     8821 ttttgatggc gttggaatt
//
