LOCUS       sequence-151420-        7671 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses calpain2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7671)
  AUTHORS   Wang Y, Zhang Y
  TITLE     Regulation of TET protein stability by calpains.
  JOURNAL   Cell Rep. 2014 Jan 30;6(2):278-84. doi: 
            10.1016/j.celrep.2013.12.031. Epub 2014 Jan 9.
  PUBMED    24412366
REFERENCE   2  (bases 1 to 7671)
  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"; doi: 
            "10.1016/j.celrep.2013.12.031"; journalName: "Cell Rep"; date: 
            "2014-01-30- 30"; volume: "6"; issue: "2"; pages: "278-84"
COMMENT     Sequence Label: p3XFlag-CAPN2
FEATURES             Location/Qualifiers
     source          1..7671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(86..105)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     296..317
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     580..602
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     594..611
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     595..611
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        772..1151
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1152..1355
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1306..1326
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3505..3570
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    3581..4203
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3780..3799)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4227..4247)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4245..4542
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4596..5390
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4650..4669)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5260..5279
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5919..6507)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5999..6018)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6678..7469,7470..7538))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7301..7320
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7539..7643)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(7670..7671,1..454)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
       61 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
      121 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
      181 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
      241 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
      301 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
      361 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
      421 gcgaatttta acaaaatatt aacgcttaca atttccattc gccattcagg ctgcgcaact
      481 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
      541 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
      601 cgacggccag tgccaagctg atctatacat tgaatcaata ttggcaatta gccatattag
      661 tcattggtta tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat
      721 atcataatat gtacatttat attggctcat gtccaatatg accgccatgt tgacattgat
      781 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      841 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      901 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      961 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     1021 atatgccaag tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     1081 cccagtacat gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg
     1141 ctattaccat ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact
     1201 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     1261 atcaacggga ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta
     1321 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagaattaag
     1381 cttatggcgg gcatcgcggc caagctggcg aaggaccggg aggcggccga ggggctgggc
     1441 tcccacgaga gggccatcaa gtacctcaac caggactacg aggcgctgcg gaacgagtgc
     1501 ctggaggccg ggacgctctt ccaggacccg tccttcccgg ccatcccctc ggccctgggc
     1561 ttcaaggagt tggggcccta ctccagcaaa acccggggca tcgagtggaa gcgccccacg
     1621 gagatctgcg ctgaccccca gtttatcatt ggaggagcca cccgcacaga catctgccaa
     1681 ggagccctag gtgactgctg gctgctggca gccattgcct ccctcacctt gaatgaagaa
     1741 atcctggctc gagtcgtccc cctaaaccag agcttccagg aaaactatgc agggatcttt
     1801 cacttccagt tctggcaata cggcgagtgg gtggaggtgg tggtggatga caggctgccc
     1861 accaaggacg gggagctgct ctttgtgcat tcagccgaag ggagcgagtt ctggagcgcc
     1921 ctgctggaga aggcatacgc caagatcaac ggatgctatg aagcactatc agggggtgcc
     1981 accactgagg gcttcgaaga cttcaccgga ggcattgctg agtggtatga gttgaagaag
     2041 ccccctccca acctgttcaa gatcatccag aaagctctgc aaaaaggctc tctccttggc
     2101 tgctccatcg acatcaccag cgccgcggac tcggaggcca tcacgtttca gaagctggtg
     2161 aaggggcacg cgtactcggt caccggagcc gaggaggttg aaagtaacgg aagcctacag
     2221 aaactgatcc gcatccgaaa tccctgggga gaagtggagt ggacagggcg gtggaatgac
     2281 aactgcccaa gctggaacac tatagaccca gaggagaggg aaaggctgac cagacggcat
     2341 gaagatggag aattctggat gtctttcagt gacttcctga ggcactattc ccgcctggag
     2401 atctgtaacc tgaccccaga cactctcacc agcgatacct acaagaagtg gaaactcacc
     2461 aaaatggatg ggaactggag gcggggctcc accgcgggag gttgcaggaa ctacccgaac
     2521 acattctgga tgaaccctca gtacctgatc aagctggagg aggaggatga ggacgaggag
     2581 gatggggaga gcggctgcac cttcctggtg gggctcattc agaagcaccg acggcggcag
     2641 aggaagatgg gcgaggacat gcacaccatc ggctttggca tctatgaggt tccagaggag
     2701 ttaagtgggc agaccaacat ccacctcagc aaaaacttct tcctgacgaa tcgcgccagg
     2761 gagcgctcag acaccttcat caacctccgg gaggtgctca accgcttcaa gctgccgcca
     2821 ggagagtaca ttctcgtgcc ttccaccttc gaacccaaca aggatgggga tttctgcatc
     2881 cgggtctttt ctgaaaagaa agctgactac caagctgtcg atgatgaaat cgaggccaat
     2941 cttgaagagt tcgacatcag cgaggatgac attgatgatg gattcaggag actgtttgcc
     3001 cagttggcag gagaggatgc ggagatctct gcctttgagc tgcagaccat cctgagaagg
     3061 gttctagcaa agcgccaaga tatcaagtca gatggcttca gcatcgagac atgcaaaatt
     3121 atggttgaca tgctagattc ggacgggagt ggcaagctgg ggctgaagga gttctacatt
     3181 ctctggacga agattcaaaa ataccaaaaa atttaccgag aaatcgacgt tgacaggtct
     3241 ggtaccatga attcctatga aatgcggaag gcattagaag aagcaggttt caagatgccc
     3301 tgtcaactcc accaagtcat cgttgctcgg tttgcagatg accagctcat catcgatttt
     3361 gataattttg ttcggtgttt ggttcggctg gaaacgctat tcaagatatt taagcagctg
     3421 gatcccgaga atactggaac aatagagctc gaccttatct cttggctctg tttctcagta
     3481 ctttgctcta gaggatcccg ggctgactac aaagaccatg acggtgatta taaagatcat
     3541 gacatcgact acaaggatga cgatgacaag tagtgatccc gggtggcatc cctgtgaccc
     3601 ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct
     3661 aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg
     3721 gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt
     3781 ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc
     3841 ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat
     3901 gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag
     3961 gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg
     4021 attacaggcg tgaaccactg ctcccttccc tgtccttctg attttaaaat aactatacca
     4081 gcaggaggac gtccagacac agcataggct acctggccat gcccaaccgg tgggacattt
     4141 gagttgcttg cttggcactg tcctctcatg cgttgggtcc actcagtaga tgcctgttga
     4201 attgggtacg cggccagctt ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4261 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4321 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4381 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4441 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4501 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggaattg atcagcttgg
     4561 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4621 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4681 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4741 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     4801 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4861 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4921 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4981 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5041 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5101 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     5161 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5221 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5281 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5341 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5401 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5461 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5521 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccgggctcg atcccctcgc
     5581 gagttggttc agctgctgcc tgaggctgga cgacctcgcg gagttctacc ggcagtgcaa
     5641 atccgtcggc atccaggaaa ccagcagcgg ctatccgcgc atccatgccc ccgaactgca
     5701 ggagtgggga ggcacgatgg ccgctttggt cgacccggac gggacgctcc tgcgcctgat
     5761 acagaacgaa ttgcttgcag gcatctcatg agtgtgtctt cccgttttcc gcctgaggtc
     5821 actgcgtgga tggagcgctg gcgcctgctg cgcgacggcg agctgctcac cacccactcg
     5881 ccaagctgga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5941 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6001 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6061 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6121 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6181 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6241 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6301 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6361 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6421 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6481 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6541 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6601 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6661 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6721 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6781 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6841 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6901 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6961 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7021 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7081 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7141 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7201 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7261 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7321 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7381 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7441 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7501 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7561 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7621 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga c
//
