LOCUS       sequence-145586-        8815 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GST fusion Sarcophilus harrisii Sacsin 90-1369.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8815)
  TITLE     Sacsin | SIRPT1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8815)
  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            "Unpublished"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-Sarcophilus harrisii
FEATURES             Location/Qualifiers
     source          1..8815
                     /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(4865..4887)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5032..5050)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5118..5222
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5223..5291,5292..6083)
                     /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(5441..5460)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6254..6842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6743..6762
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6996..7013
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        7086..7163
                     /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             7164..8246
                     /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(7183..7202)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    8259..8280
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8295..8300,8301..8318,8319..8325)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8333..8349
                     /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     8357..8373
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8357..8373
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(8369..8815,1..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(8389..8406)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8389..8405)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8398..8420)
                     /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 gtcagaccac gccacctctg gtagattttc tgaaggacat cctgcgtcgt tacccagaag
     1021 gtggtcaaat cctgaaggaa ctgatccaga acgctgaaga cgctggtgct actgaagtaa
     1081 agttcctgta cgacgaaact cagtacggta aggaaactct gtggtctaag gacatggaac
     1141 agtaccaggg ttctgcactg tatgtataca acaacgcggt attcacgcct gaagactggc
     1201 atggtatcca agagatcgca cgtagccgta agaaggacga ccctctgaaa gtaggtcgtt
     1261 tcggtatcgg tttcaactct gtgtaccaca tcaccgacgt cccttgtatc ttctctggtg
     1321 accaaatcgg tatgctggac cctcatcaaa ccctgttcgg tcctcatgaa tctggtcaat
     1381 gctggaacct gaaggacgac tctaaggaaa tcagcgagtt cagcgaccaa ttcgcacctt
     1441 tcatcggtgt gttcggttct acgaaggaga ctttcctgaa cggtaacttc ccgggtactt
     1501 tcttccgctt ccctctgcgt ctgcaaccat ctcaactgtc taacaacctg tacaacaagc
     1561 agaagatcct ggagctgttc gacagcttca aggcagacgc tgacactgtc ctgctgttcc
     1621 tgaagtctgt acaagacgta agcctgcatg tacgtgaagc agacggtact gaaaagctga
     1681 tcttccgcgt cactgcatct gaaaacaagg cgctgaagca tgagcgtcca aactctatca
     1741 agatcctggg tacggcaatc tctaactact gcaagaaggt cccgagcaac agcatcacgt
     1801 gtgtcactta ccacatcaac atcgtcctgg aggacgaaag cactaaggac gctcaaaaga
     1861 cctcctggct ggtctgtaac tctgtcggtg gtcgtggtat ttgtaacgag ctggactgtc
     1921 tggcagatga tctgaaattc gtgccaacta tcggtatcgc aatgtccctg tcctctagcg
     1981 aagagcagac tggtgcaatc gcagatttta gcggtaaagc cttctgcttc ctgccactgc
     2041 caccaggtga agaatctaaa accggtctgc cagttcatgt ctctggtttc ttcggtctga
     2101 ctgacaaccg tcgttctatc aaatggcgtg agctggacca gtggcgtgat ccagcagcac
     2161 tgtggaatga actgctgatt gttaacgtcg ttccaaaagc ttacgcaact ctggtactgg
     2221 actctatcaa acgcctggag actgagaaaa actccgactt cccgctgtct gtagagatca
     2281 tctacaaact gtggccggag ctgtctcgtg tgaaagtgca ttggcagccg atcatcgaac
     2341 cgctgtttaa agagctgttc cagaacgcag tgatctacag catcagcaac cactggatca
     2401 aactggagca ggcatacttc tccgagctgg acgaatctct ggaatacacg aaagccgtgc
     2461 tgaactacct gcagtcttct ggcaaacaga tcgtgaaagt gccgactaac atctccaacg
     2521 ctgtgcagct gactgcttct tctgctaaac cggtgaaaaa agtgaccccg gctctggttc
     2581 gtcaggtact gaaaaaacac ggccattctg gcaccgctgc tgaaaaactg catctgctgg
     2641 aattcgtgct gtctgatgct gcttaccatg aactgctggg cctggatctg ctgccgctgc
     2701 agaatggcaa ttttattccg ttttctagca gcgtgtctga ccaggacgtt atctacatca
     2761 ccagcgaaga ctacccgcgt tctctgttcc cgggcctgga aggccgtatt attctggacg
     2821 atctgaaacc gcagattctg gcttctctga aagaagctgc tcagacccgt ggccgtccgt
     2881 gtacccaact gcagctgctg aattctgaac gttttgctcg tctgattaaa gaagttatga
     2941 atgcgttttg gccgggccgt gaactggttg ttcagtggca tccgtttgcg gaagattctc
     3001 atcacccgtc tgttgtttgg ctgaaaatgg tttggaaaaa cctgtacatc cacttctccg
     3061 aagacctgtc tctgttcgac gaaatgccgc tgatcccgaa aaccatcctg gaaaaatccc
     3121 agaccttcat ggaactgatc cgtctgcgta tcccgtctcc ggttattctg gatgacgaat
     3181 ctgaagcgca gctgccggaa tttctggccg atattgtaca gaacctgggc ggcattgttc
     3241 tgaaaaaact ggacccgtct atccagcacc cgctgattaa aaaatacgtg cacgcgccgc
     3301 tgccgtctgc ggttctgcag attatggaaa aaatgtctct gcagaaactg tgtaaccagg
     3361 tggcgtctct gctgccgacc cacaaagatg ccctgcgcaa atttctggcg agcctgacgg
     3421 atattaatga aaaagaaaaa cgcatcatcc aggaactgac catcttcaaa cgcatcaacc
     3481 acagctcctc ccagggcatt agctccttta ccaaactgaa aggctgcaaa gtgctgcacc
     3541 acaccgcgaa actgccgccg gatctgcgcc tgtccattcc ggtaattgat aattccgatg
     3601 aagccaccat tcgcctggcc aacatgctga aagtagaaca gctgaaaagc accggctgcc
     3661 tgaaatttat cctgaaagat atcgaaaacg gcttctatag ccacgaagag accacccagc
     3721 tgatgctgtg gattctggaa cacctgtcca gcctgaaaaa tgaaaacccg cgcgttattg
     3781 actgggttat gccgctgaaa ttcatccaga tctcccaggg ccacattgtt tccgccaatg
     3841 aactgttcga ctccgatatc tccattctgc aggacctgtt ctatgccgaa gaagagatct
     3901 ccttcccgcc gtccattttc accagcagcc tggatattct gcactccctg cgccagattg
     3961 gcctgaaaaa cgaaacctcc ctgaaagaaa aagatatcat ccaggtggcg aaaaaaatcg
     4021 aggagctgca ggtgtccagc agcccgaacc tggatgccct gctgaaaaaa gcgaaaaccc
     4081 tgctgatgat tctgaacaaa aaccacaccc tgctgcagtc cccggaagcc aaagcgaccc
     4141 tgaaaaaaat taaatgggtg ccgacctgca aagaacgccc gccgaactat ccgggcagcc
     4201 tgatttggaa aggcgatctg tgcaacctgt gctccccgcc ggaaatgtgc gatgtatccc
     4261 acgcgaccct ggttggctcc agcgttccgc tggttgaatc cattcacgcg aacctggaaa
     4321 aagcgctggg catttttacc aaaccgtcca ttaacgcggt tctgaaacac tttaaaattg
     4381 ttgttgattg gcactcctcc aaaacgtttt ccgatgaaga ttattatcag tttcagcaca
     4441 tcctgctgga aatctatggc tttatgcacg atcacctgga tgaaggcaaa gaagccttca
     4501 aagcgctgaa attcccgtgg gtttggaccg gcaaaaaatt ctgcccgctg gcgcaggcgg
     4561 ttatcaaacc gatccacgat ctggaactgc agccgtatct gcactccgtt ccgaaaacca
     4621 tggcgaaatt ccaccagctg ttcaaagttt gcggcagcat cgaagaactg accagcgatc
     4681 acatcagcat ggttatccag aaagtttatc tgaaaagcga tcaggaactg tccgaagagg
     4741 agtccaaaca gaacctgcac ctgatgctga acatcatccg ctggctgtat agctcctaac
     4801 tcgagcggcc gcatcgtgac tgactgacga tctgcctcgc gcgtttcggt gatgacggtg
     4861 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     4921 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
     4981 tgacccagtc acgtagcgat agcggagtgt ataattcttg aagacgaaag ggcctcgtga
     5041 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     5101 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5161 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5221 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5281 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5341 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5401 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5461 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5521 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5581 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5641 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5701 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5761 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5821 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5881 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5941 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6001 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6061 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     6121 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6181 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6241 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6301 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6361 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     6421 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6481 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6541 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6601 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6661 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6721 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6781 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6841 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6901 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6961 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     7021 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcataa
     7081 attccgacac catcgaatgg tgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
     7141 agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg
     7201 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
     7261 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
     7321 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
     7381 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
     7441 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
     7501 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
     7561 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
     7621 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
     7681 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
     7741 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
     7801 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
     7861 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
     7921 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
     7981 acgacgatac cgaagacagc tcatgttata tcccgccgtc aaccaccatc aaacaggatt
     8041 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
     8101 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca
     8161 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     8221 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     8281 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     8341 ggataacaat ttcacacagg aaacagctat gaccatgatt acggattcac tggccgtcgt
     8401 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     8461 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     8521 gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt ccggcaccag aagcggtgcc
     8581 ggaaagctgg ctggagtgcg atcttcctga ggccgatact gtcgtcgtcc cctcaaactg
     8641 gcagatgcac ggttacgatg cgcccatcta caccaacgta acctatccca ttacggtcaa
     8701 tccgccgttt gttcccacgg agaatccgac gggttgttac tcgctcacat ttaatgttga
     8761 tgaaagctgg ctacaggaag gccagacgcg aattattttt gatggcgttg gaatt
//
