LOCUS       sequence-151411-        7721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses calpain1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7721)
  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 7721)
  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-CAPN1
FEATURES             Location/Qualifiers
     source          1..7721
                     /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             3555..3620
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    3631..4253
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3830..3849)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4277..4297)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4295..4592
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4646..5440
                     /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(4700..4719)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5310..5329
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5969..6557)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6049..6068)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6728..7519,7520..7588))
                     /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     7351..7370
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7589..7693)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(7720..7721,1..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 cttgcggccg cgaattcacc atgtcggagg agatcatcac gccggtgtac tgcactgggg
     1441 tgtcagccca agtgcagaag cagcgggcca gggagctggg cctgggccgc catgagaatg
     1501 ccatcaagta cctgggccag gattatgagc agctgcgggt gcgatgcctg cagagtggga
     1561 ccctcttccg tgatgaggcc ttccccccgg taccccagag cctgggttac aaggacctgg
     1621 gtcccaattc ctccaagacc tatggcatca agtggaagcg tcccacggaa ctgctgtcaa
     1681 acccccagtt cattgtggat ggagctaccc gcacagacat ctgccaggga gcactggggg
     1741 actgctggct cttggcggcc attgcctccc tcactctcaa cgacaccctc ctgcaccgag
     1801 tggttccgca cggccagagc ttccagaatg gctatgccgg catcttccat ttccagctgt
     1861 ggcaatttgg ggagtgggtg gacgtggtcg tggatgacct gctgcccatc aaggacggga
     1921 agctagtgtt cgtgcactct gccgaaggca acgagttctg gagcgccctg cttgagaagg
     1981 cctatgccaa ggtaaatggc agctacgagg ccctgtcagg gggcagcacc tcagagggct
     2041 ttgaggactt cacaggcggg gttaccgagt ggtacgagtt gcgcaaggct cccagtgacc
     2101 tctaccagat catcctcaag gcgctggagc ggggctccct gctgggctgc tccatagaca
     2161 tctccagcgt tctagacatg gaggccatca ctttcaagaa gttggtgaag ggccatgcct
     2221 actctgtgac cggggccaag caggtgaact accgaggcca ggtggtgagc ctgatccgga
     2281 tgcggaaccc ctggggcgag gtggagtgga cgggagcctg gagcgacagc tcctcagagt
     2341 ggaacaacgt ggacccatat gaacgggacc agctccgggt caagatggag gacggggagt
     2401 tctggatgtc attccgagac ttcatgcggg agttcacccg cctggagatc tgcaacctca
     2461 cacccgacgc cctcaagagc cggaccatcc gcaaatggaa caccacactc tacgaaggca
     2521 cctggcggcg ggggagcacc gcggggggct gccgaaacta cccagccacc ttctgggtga
     2581 accctcagtt caagatccgg ctggatgaga cggatgaccc ggacgactac ggggaccgcg
     2641 agtcaggctg cagcttcgtg ctcgccctta tgcagaagca ccgtcgccgc gagcgccgct
     2701 tcggccgcga catggagact attggcttcg cggtctacga ggtccctccg gagctggtgg
     2761 gccagccggc cgtacacttg aagcgtgact tcttcctggc caatgcgtct cgggcgcgct
     2821 cagagcagtt catcaacctg cgagaggtca gcacccgctt ccgcctgcca cccggggagt
     2881 atgtggtggt gccctccacc ttcgagccca acaaggaggg cgacttcgtg ctgcgcttct
     2941 tctcagagaa gagtgctggg actgtggagc tggatgacca gatccaggcc aatctccccg
     3001 atgagcaagt gctctcagaa gaggagattg acgagaactt caaggccctc ttcaggcagc
     3061 tggcagggga ggacatggag atcagcgtga aggagttgcg gacaatcctc aataggatca
     3121 tcagcaaaca caaagacctg cggaccaagg gcttcagcct agagtcgtgc cgcagcatgg
     3181 tgaacctcat ggatcgtgat ggcaatggga agctgggcct ggtggagttc aacatcctgt
     3241 ggaaccgcat ccggaattac ctgtccatct tccggaagtt tgacctggac aagtcgggca
     3301 gcatgagtgc ctacgagatg cggatggcca ttgagtcggc aggcttcaag ctcaacaaga
     3361 agctgtacga gctcatcatc acccgctact cggagcccga cctggcggtc gactttgaca
     3421 atttcgtttg ctgcctggtg cggctagaga ccatgttccg atttttcaaa actctggaca
     3481 cagatctgga tggagttgtg acctttgact tgtttaagtg gttgcagctg accatgtttg
     3541 caggatcccg ggctgactac aaagaccatg acggtgatta taaagatcat gacatcgact
     3601 acaaggatga cgatgacaag tagtgatccc gggtggcatc cctgtgaccc ctccccagtg
     3661 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     3721 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     3781 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     3841 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     3901 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     3961 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     4021 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     4081 tgaaccactg ctcccttccc tgtccttctg attttaaaat aactatacca gcaggaggac
     4141 gtccagacac agcataggct acctggccat gcccaaccgg tgggacattt gagttgcttg
     4201 cttggcactg tcctctcatg cgttgggtcc actcagtaga tgcctgttga attgggtacg
     4261 cggccagctt ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4321 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4381 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4441 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4501 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4561 ctattccaga agtagtgagg aggctttttt ggaggaattg atcagcttgg gatctgatca
     4621 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4681 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4741 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4801 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     4861 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4921 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4981 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5041 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5101 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5161 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5221 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5281 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5341 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5401 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5461 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5521 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5581 ggggatctca tgctggagtt cttcgcccac cccgggctcg atcccctcgc gagttggttc
     5641 agctgctgcc tgaggctgga cgacctcgcg gagttctacc ggcagtgcaa atccgtcggc
     5701 atccaggaaa ccagcagcgg ctatccgcgc atccatgccc ccgaactgca ggagtgggga
     5761 ggcacgatgg ccgctttggt cgacccggac gggacgctcc tgcgcctgat acagaacgaa
     5821 ttgcttgcag gcatctcatg agtgtgtctt cccgttttcc gcctgaggtc actgcgtgga
     5881 tggagcgctg gcgcctgctg cgcgacggcg agctgctcac cacccactcg ccaagctgga
     5941 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6001 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6061 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6121 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6181 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6241 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6301 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6361 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6421 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6481 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6541 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6601 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6661 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6721 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6781 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6841 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6901 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6961 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7021 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7081 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7141 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7201 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7261 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7321 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7381 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7441 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7501 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7561 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7621 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7681 taggggttcc gcgcacattt ccccgaaaag tgccacctga c
//
