LOCUS       sequence-412141-        8037 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAPN2 protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8037)
  TITLE     TREAT-AD plasmids Jun 2023
REFERENCE   2  (bases 1 to 8037)
  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: CAPN2A-c002
FEATURES             Location/Qualifiers
     source          1..8037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(71..1153)
                     /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     1115..1134
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1154..1231)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1437..1456
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1540..1559
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1540..1558
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1559..1583
                     /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)."
     RBS             1598..1620
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1639..1656
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1666..1686
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3808..3852
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             4441..4458
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4511..4529)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4525..4572
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4609..5064
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4696..4715)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4906..4927
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5156..5971)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5156..5971)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5879..5898
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5963..5982
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6093..6681
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6582..6601
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6835..6852
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6867..7006
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6932..6951)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7092..7114
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7108..7299)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
       61 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
      121 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
      181 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
      241 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
      301 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac
      361 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
      421 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
      481 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
      541 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
      601 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
      661 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
      721 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
      781 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
      841 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
      901 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
      961 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     1021 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     1081 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     1141 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     1201 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
     1261 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
     1321 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     1381 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     1441 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     1501 cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg
     1561 aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga
     1621 tataccatgg gcagcagcca tcatcatcat catcacagca gcggcgagaa tctttatttt
     1681 cagggccata tggctggaat agcggcaaag ctagctaaag atcgtgaggc tgcggaaggt
     1741 ctgggttctc atgatcgtgc gatcaaatac ctgaaccagg actacgaagc tttgagaaac
     1801 gagtgcctgg aagctggtac attgtttcag gatccgtcct tcccggcaat cccgagcgcg
     1861 ctgggcttta aggagctggg cccatacagc agcaaaacca gaggtattga atggaagcgc
     1921 ccaactgaaa tttgcgctga tccgcaattt attattggcg gtgccacccg caccgatatc
     1981 tgccaaggtg cgcttgggga ttgttggctg ttggcagcca tcgcaagttt gaccctgaac
     2041 gaggagatcc tggctcgtgt ggtgccgctg aaccagtcct tccaagaaaa ttacgcgggg
     2101 atatttcact ttcagttttg gcagtatggc gaatgggttg aggtcgttgt ggacgaccgg
     2161 ctgccgacga aggacggtga actgttattc gtgcattccg cggagggctc tgagttctgg
     2221 tcagcccttc tggaaaaagc gtacgcgaaa atcaatggct gctatgaggc actgagcggt
     2281 ggtgcgacca ccgagggctt cgaggacttc acgggtggca tagcggagtg gtatgaactc
     2341 aagaagccac cgcctaatct ttttaagatc atccaaaaag cgttacaaaa gggctccctg
     2401 ctcgggtgca gcattgacat cactagcgcg gcggacagcg aagcgattac cttccaaaag
     2461 ttggttaaag gtcatgcata cagcgtcacc ggtgctgagg aggtggaaag caatggttct
     2521 ctgcaaaagc tgatccgaat tcgtaatccg tggggtgaag tggaatggac tggtcgttgg
     2581 aacgataact gcccgagctg gaacaccatt gacccggaag aacgcgagcg tttaacccgt
     2641 cgtcacgaag acggcgagtt ctggatgtcg ttcagcgact tcctgcgtca ttatagccgc
     2701 ctagagatct gcaacctgac tccggatacc ctcacgagcg acacctataa aaaatggaag
     2761 ttgacgaaga tggatggcaa ttggcgtcgt ggctctacgg ccggcggctg ccgtaactac
     2821 cccaacacct tttggatgaa cccgcagtac ctgattaagt tagaagaaga agacgaggac
     2881 gaagaagacg gcgagtccgg ctgcaccttc ctggtcggtc tgatccaaaa gcaccgtcgt
     2941 cgccagcgta aaatgggtga agacatgcac accatcggct tcggtattta tgaggtgccg
     3001 gaagagttga gtggtcagac caatattcat ctctctaaga acttcttttt gacgaaccgc
     3061 gcgagagagc gcagcgatac cttcatcaat ttgcgtgagg tcttgaaccg ctttaagcta
     3121 ccgccgggtg agtatattct ggttccgtcg accttcgaac cgaataaaga cggtgatttt
     3181 tgtatccgtg tattttctga aaaaaaggcg gattatcagg cagttgatga cgagatcgag
     3241 gctaatctgg aggagttcga tatcagcgag gacgatattg acgacggttt ccgccgcctg
     3301 tttgcacagc tggcgggtga ggatgcagag attagcgcgt ttgagctgca aactatcctg
     3361 cgccgtgttc tggccaaacg tcaggacatt aaatccgatg gcttctccat cgaaacctgt
     3421 aaaatcatgg ttgacatgct ggatagcgat ggcagcggta agctgggcct caaagaattc
     3481 tatatcctgt ggaccaagat tcaaaagtac cagaaaatct accgtgaaat tgatgttgat
     3541 cgctccggta cgatgaacag ctatgaaatg cgcaaagcgc tggaagaggc cggcttcaaa
     3601 atgccgtgtc agcttcacca ggttatcgtg gcgcgttttg ccgatgacca gctgattatc
     3661 gacttcgaca actttgttcg ttgcctggtg cgtttggaga ccctgtttaa aattttcaag
     3721 cagttggacc cggagaacac cggcaccatc gagctggacc tgattagctg gctgtgtttt
     3781 tcggtgctgg gctctaaggg tggttacggt ttgaacgata ttttcgaagc tcaaaaaatt
     3841 gaatggcacg agtaataata atttaaggag atatacatga gtcactattc aaacatagaa
     3901 gctaatgaga gcgaagaggt gcgccagttt agacgtttat tcgcccaatt ggcgggcgat
     3961 gatatggaag tcagcgcaac cgaactgatg aacatcctga ataaagtagt tacccgtcat
     4021 ccggatttga aaaccgacgg cttcggtatt gatacgtgcc gttctatggt tgctgtgatg
     4081 gacagcgata ctacgggtaa actgggtttc gaggagttca agtacctgtg gaataacatc
     4141 aaacgttggc aggcgattta taagcagttc gacaccgatc gttccggcac catctgttcc
     4201 tcggaactgc cgggtgcgtt tgaagcggcg ggcttccact tgaacgagca cctgtataac
     4261 atgattatcc gtcgctactc tgacgaaagc ggtaatatgg attttgacaa ctttatcagc
     4321 tgcctggttc gtctggacgc tatgttccgc gcatttaagt ccctcgacaa ggacggcacc
     4381 ggtcagattc aagtgaacat ccaagagtgg ctgcagttga ccatgtacag ctaactcgag
     4441 caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg
     4501 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     4561 aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta
     4621 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4681 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4741 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4801 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4861 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4921 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4981 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5041 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5101 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga
     5161 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     5221 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     5281 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     5341 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     5401 ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat
     5461 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     5521 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     5581 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     5641 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     5701 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     5761 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     5821 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     5881 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag
     5941 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     6001 cagacagttt tattgttcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6061 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6121 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6181 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     6241 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6301 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6361 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6421 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6481 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6541 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6601 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6661 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6721 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6781 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     6841 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     6901 ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     6961 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     7021 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     7081 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     7141 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     7201 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     7261 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     7321 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     7381 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     7441 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     7501 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     7561 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     7621 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     7681 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     7741 caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc
     7801 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     7861 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     7921 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     7981 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaa
//
