LOCUS       sequence-145578-        8911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GST fusion  Nelumbo nucifera Sacsin 1-1313.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8911)
  TITLE     Sacsin | SIRPT1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8911)
  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            "Unpublished"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-Nelumbo nucifera
FEATURES             Location/Qualifiers
     source          1..8911
                     /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(4961..4983)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5128..5146)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5214..5318
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5319..5387,5388..6179)
                     /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(5537..5556)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6350..6938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6839..6858
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7092..7109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        7182..7259
                     /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             7260..8342
                     /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(7279..7298)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    8355..8376
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8391..8396,8397..8414,8415..8421)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8429..8445
                     /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     8453..8469
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8453..8469
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(8465..8911,1..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(8485..8502)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8485..8501)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8494..8516)
                     /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 catatggagt
      961 ctagcggtac ggacagcttc ctgctggagg atttcggtca aaaggtcgac ctgactcgtc
     1021 gtatccgtga agttctggta aactaccctg agggtactac cgtcctgaag gaactgatcc
     1081 agaacgctga cgacgctggt gcaactaagg tctgtctgtg tctggaccgt cgtgttcatg
     1141 gtgttggttc tctgctgtct tctaagctgg cagaatggca aggtcctgct ctgctggctt
     1201 acaacaacgc agaattcacc gacgatgatt tcgtatccat cagccgtatc ggtgacagca
     1261 agaagcaagg tcaggcatgg aagactggtc gtttcggtgt aggtttcaac agcgtctacc
     1321 acctgactga cctgcctagc ttcgtaagcg gtaagtacgt cgtcctgttc gacccacaag
     1381 gtatctacct gcctaacatc tccgtcgcaa accctggtaa gcgtatcgaa tacgtcaaca
     1441 gcagcgcaat gagcctgtac aaggaccagt tcctgcctta ctgcactttc ggttgtgaca
     1501 tgaagcgccc tttccatggt accctgttcc gtttccctct gcgtaacgca gaccaagcag
     1561 caactagcaa gctgagccgt caagcatacc tggaagacga catcagcagc atgttcctgc
     1621 aactgtacga ggaaggtgtc ctgactctgc tgttcctgaa gaacgtgatc agcgtggaaa
     1681 tgtacctgtg ggactccggt gctctggaac cacagaagat ctacagctgt tccgtcaaca
     1741 gcgctaacga agacactatc tggcaccgtc aggcactgct gcgtctgtct aactctatcg
     1801 tgagcagcaa catcgaaatg gactccttca gcctggactt cctgagcgaa aaagtggccg
     1861 gtaactctct ggaaaaaaaa gtggacacct tccacatcgt gcaggctatg gcgccggctt
     1921 cttctaaaat cggtgccttc gcagcatctg cagcaaaaga gtacgacatc cacctgctgc
     1981 catgggcatc tgtggcagct tgtatctctg actctctgct ggaagacaac gtgctgaaac
     2041 acggtcgtgc attctgtttc ctgccactgc cagtgcgtac tggtatggca gtgcagatca
     2101 acggttactt cgaggtgtct tctaaccgcc gttctatctg gtacggtgac gacatggacc
     2161 gtggtggtaa actgcgttct gactggaata tgctgctgct ggaggacgtt gtgtctccag
     2221 ctttcactcg tctgctgctg ggtgttcgtt ctctgctggg tccagaaaaa ctgtactaca
     2281 acctgtggcc atctggttct ttcgaggagc catggaactt cctggtggaa tgcatctacc
     2341 gtaacgtggg taactcccca gtgctgtact ctgacgtggg tggcggcaaa tgggtgtctc
     2401 cggctgaagc tttcatccac gacgaactgt tcacccgttc taaagagctg ggtgaggctc
     2461 tgctgctggt tggtatgtct atcgtgtgtc tgccgaactt cctgtctgac atgctgctgc
     2521 gttactcttc ctctttcatc cagcgcgtga tcaccccgtc taccgttcgt tgctttctgc
     2581 gtgaatgtaa aaccctgatc tccctgtcca aattctaccg cctggtgctg ctggaatact
     2641 gcctggagga cctgatcgac gaagacgtgg gtcgtgatgc ttctgaactg ccgctgattc
     2701 cgctggcttc tggtgaattt gcttctttct ctgaggcttc taaaggcttc ccgtacttca
     2761 tctgctccga gctggaatac atgctgctgt accagatccc ggaccgtgtt atcgaccgta
     2821 acatcccgcc ggatatcctg acccgtattt ctgctatcgc taaatcctcc aactccaaca
     2881 tcgtgacctt caacgacaaa tacttcctgc agctgttccc gaaattcttc ccgtccgagt
     2941 ggaaatacaa aaaccgcgtt ctgtggaacc cggagtcctc cccgatccac ccgtcctcct
     3001 cttggttcct gctgttctgg cagtacctgc gtgaccagtg tgaaaacctg tctctgttct
     3061 gcgactggcc gattttcccg tctacgtctg gtcatctgta ccgtgctctg cgtcatctga
     3121 aactgatcga cgctgaaaaa ctgccgggta aaatgcgcac cctgctggct aaaatcggct
     3181 gtcgtatcct gtctccgaac tacggcgttg aacactggga actgtctcac tacgttcacg
     3241 gcgcggatgg cgcgggcgtt ctggaagcga tttttgatgt tgtttcttct aacgaagacc
     3301 tgaaacacct gttcaacgac aacctgggcg ttgacgaaaa agacgagctg cgtcactttc
     3361 tgctggaccc gaaatggtac atcggcgatt gcatcctgga gctgcacatc cagaaatgca
     3421 aaatgctgcc gatctataaa gttcacgatg gcgagtccat ccagaccttc cacttctccg
     3481 atctggagaa cccgatcaaa tatctgccgc cgtccgatat cccgaaatat ctgctgggcg
     3541 aggaatttat ctgcagcctg tccgataccg aagaaaaaat cctgctgggc tattatggca
     3601 tcgaacagat gggcaaagcc tgcttttata aacagcaggt tctgaaccgc atctgcgaac
     3661 tgcagccgga agtacgcgat cgtgttatgc tgtccattct gcaggatctg ccgcagctgt
     3721 gcgccgaaga aacgtccctg cgcgatagcc tgcgcaaact ggaatttgtt ccgacgctga
     3781 gcggcatcct gaaatgcccg gatgcgctgt atgatccgcg caacgaagaa ctgtatgcgc
     3841 tgctggaaga ttccgattcc tatccgtatg gcctgtttca ggaatccggc gccctggata
     3901 tgctgatcgg cctgggcctg cgtacctttg tttctccgga aaccattatt cagagcgccc
     3961 gtcagattga actgatgatg cacaaagatc agcagaaagc gcacgtaaaa ggcaaagcgc
     4021 tgctgtccta tctggaagta aacgcggtta aatggagctt taacctgctg aacgatggca
     4081 aacgccgcat gaaccgtctg tttagccagg ttgcgaccag ctttaaaccg cgcaattccg
     4141 aaattgatct ggaaaaattt tggaatgatc tgcgcatgat ttgctggtgc ccggttctgg
     4201 tagcggcgcc gtatccgtcc ctgccgtggc cgtccatttc cagcatggtt gcgccgccga
     4261 aactggttcg cctgccggcg gatatgtggc tggtatctgc gtctctgcgt attctggatg
     4321 gcgaatgctc cagcaccgcg ctgagctcca gcctgggctg gtccaccacg ccgggcggct
     4381 ccattctgag cgcccagctg ctggaactgg gtaaaaataa tgaactggtt caggataaag
     4441 ttctgcgtca ggaactggcc ctggcgatgc cgaaaattta ttccattctg acctccatga
     4501 ttggctccga tgaaatggat attgtaaaag cgattctgga aggctgtcgt tggatttggg
     4561 ttggcgatgg ctttgcgacc ctggatgaag tagtactgaa tggcccgctg catctggttc
     4621 cgtatattcg tgtaattccg gtagatctgg ccgtttttcg cgaactgttt ctggaactgg
     4681 gcattcgcga atttctgaaa ccggaagatt atgccggcat tctgcaccgc atggcggccc
     4741 gcaaaggctg caatccgctg gatgcgcatg aactgcgcgc ggcggtactg attgtacagc
     4801 atctggccga agcgcatttt caggataaac acaatgaaat ttatctgccg gatgttagca
     4861 gccgcctgtt tagcgcgacc gatctggttt attaactcga gcggccgcat cgtgactgac
     4921 tgacgatctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     4981 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     5041 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     5101 gagtgtataa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5161 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5221 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5281 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5341 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5401 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5461 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5521 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     5581 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5641 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5701 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5761 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5821 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt
     5881 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5941 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6001 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6061 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6121 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6181 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6241 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6301 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6361 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6421 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6481 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6541 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6601 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6661 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6721 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6781 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6841 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6901 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6961 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     7021 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     7081 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     7141 tccttacgca tctgtgcggt atttcacacc gcataaattc cgacaccatc gaatggtgca
     7201 aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg
     7261 tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt
     7321 cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg
     7381 cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt
     7441 cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg
     7501 cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac
     7561 gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg
     7621 ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca
     7681 ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt
     7741 tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc
     7801 aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct
     7861 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
     7921 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
     7981 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
     8041 ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat
     8101 gttatatccc gccgtcaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
     8161 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg
     8221 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
     8281 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     8341 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     8401 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     8461 agctatgacc atgattacgg attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     8521 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     8581 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     8641 gcgctttgcc tggtttccgg caccagaagc ggtgccggaa agctggctgg agtgcgatct
     8701 tcctgaggcc gatactgtcg tcgtcccctc aaactggcag atgcacggtt acgatgcgcc
     8761 catctacacc aacgtaacct atcccattac ggtcaatccg ccgtttgttc ccacggagaa
     8821 tccgacgggt tgttactcgc tcacatttaa tgttgatgaa agctggctac aggaaggcca
     8881 gacgcgaatt atttttgatg gcgttggaat t
//
