LOCUS       sequence-45113-e        5423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control for pCMV-calpainsensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5423)
  AUTHORS   Stockholm D, Bartoli M, Sillon G, Bourg N, Davoust J, Richard I
  TITLE     Imaging calpain protease activity by multiphoton FRET in living 
            mice.
  JOURNAL   J Mol Biol. 2005 Feb 11;346(1):215-22. Epub 2004 Dec 16.
  PUBMED    15663939
REFERENCE   2  (bases 1 to 5423)
  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: "J Mol 
            Biol."; date: "2005-02-11"; volume: "346(1)"; pages: "215-22. Epub 
            2004 Dec 16"
COMMENT     Sequence Label: pCMV_eYFP-eCFP
FEATURES             Location/Qualifiers
     source          1..5423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    422..543
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(459..478)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     513..532
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(550..1005)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(687..708)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     899..918
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1032..1136
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1138..1495
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1138..1329
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1161..1181)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1346..1481
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1530..2324
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1530..2324
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1584..1603)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2194..2213
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2512..2531)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2556..2603
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2932..3520
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3421..3440
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3695..3998
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3999..4202
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4153..4173
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4241..4250
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4247..4249,4250..4252,4253..4963)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      4997..5006
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5003..5005,5006..5008,5009..5423,1..299)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
ORIGIN
        1 agctggagta caactacatc agccacaacg tctatatcac cgccgacaag cagaagaacg
       61 gcatcaaggc caacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
      121 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
      181 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
      241 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa
      301 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
      361 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
      421 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
      481 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
      541 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
      601 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
      661 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
      721 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
      781 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
      841 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
      901 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
      961 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     1021 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1081 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     1141 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     1201 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     1261 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     1321 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     1381 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     1441 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     1501 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     1561 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     1621 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     1681 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     1741 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     1801 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     1861 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     1921 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     1981 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     2041 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     2101 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     2161 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     2221 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     2281 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     2341 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     2401 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     2461 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     2521 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     2581 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     2641 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     2701 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     2761 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     2821 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     2881 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2941 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3001 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3061 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3121 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3181 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3241 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3301 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3361 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3421 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3481 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3541 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     3601 tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg
     3661 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     3721 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     3781 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     3841 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     3901 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     3961 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     4021 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     4081 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     4141 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     4201 ctggtttagt gaaccgtcag atccgctagc gctaccggtc gccaccatgg tgagcaaggg
     4261 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     4321 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     4381 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt
     4441 cggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     4501 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     4561 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     4621 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     4681 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     4741 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     4801 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca
     4861 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     4921 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtccggac tcagatctcg
     4981 aggggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     5041 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     5101 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     5161 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tggggcgtgc agtgcttcag
     5221 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     5281 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     5341 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     5401 ggacggcaac atcctggggc aca
//
