LOCUS       sequence-42429-e        5424 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a fusion between the Lck domain and cytosolic GCaMP5G. 
            This construct targets GCaMP5G to the plasma membrane.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5424)
  AUTHORS   Akerboom J, Chen TW, Wardill TJ, Tian L, Marvin JS, Mutlu S, 
            Calderon NC, Esposti F, Borghuis BG, Sun XR, Gordus A, Orger MB, 
            Portugues R, Engert F, Macklin JJ, Filosa A, Aggarwal A, Kerr RA, 
            Takagi R, Kracun S, Shigetomi E, Khakh BS, Baier H, Lagnado L, Wang 
            SS, Bargmann CI, Kimmel BE, Jayaraman V, Svoboda K, Kim DS, 
            Schreiter ER, Looger LL
  TITLE     Optimization of a GCaMP calcium indicator for neural activity 
            imaging.
  JOURNAL   J Neurosci. 2012 Oct 3;32(40):13819-40. doi: 
            10.1523/JNEUROSCI.2601-12.2012.
  PUBMED    23035093
REFERENCE   2  (bases 1 to 5424)
  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: 
            "10.1523/JNEUROSCI.2601-12"; journalName: "J Neurosci"; date: 
            "2012-10-3- 3"; volume: "32"; issue: "40"; pages: "13819-40"
COMMENT     Sequence Label: pN1 Lck-GCaMP5G
FEATURES             Location/Qualifiers
     source          1..5424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      615..624
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      732..741
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(738..746,747..764,765..767,768..800,801..803,804..827,
                     828..854,855..911,912..917,918..1187,1188..1205,1206..1640,
                     1641..1646,1647..2090)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             join(738..746,747..764,765..767,768..800,801..803,804..827,
                     828..854,855..911,912..917,918..1187,1188..1205,1206..1640,
                     1641..1646,1647..2090)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      834..843
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1124..1145
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1251..1272)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1512..1531)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2212..2333
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2249..2268)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2303..2322
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2340..2795)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2477..2498)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2689..2708
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2822..2926
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2928..3285
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2928..3119
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2951..2971)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3136..3271
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3320..4114
                     /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             3320..4114
                     /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(3374..3393)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3984..4003
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4302..4321)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4346..4393
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4722..5310
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5211..5230
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctgccacc atgggctgtg gctgcagctc aaaccctgaa gatgactgga
      661 tggagaacat tgacgtgtgt gagaactgcc attatccccc gccgaagctt cgaattgatc
      721 caccggatct cgccaccatg ggttctcatc atcatcatca tcatggtatg gctagcatga
      781 ctggtggaca gcaaatgggt cgggatctgt acgacgatga cgataaggat ctcgccacca
      841 tggtcgactc atcacgtcgt aagtggaata agacaggtca cgcagtcaga gctataggtc
      901 ggctgagctc actcgagaac gtctatatca aggccgacaa gcagaagaac ggcatcaagg
      961 cgaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc taccactacc
     1021 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagcg
     1081 tgcagtccaa actttcgaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1141 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc ggtaccggag
     1201 ggagcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1261 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggtgag ggcgatgcca
     1321 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     1381 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     1441 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacatccag gagcgcacca
     1501 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1561 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1621 ggcacaagct ggagtacaac ctgccggacc aactgactga agagcagatc gcagaattta
     1681 aagaggcttt ctccctattt gacaaggacg gggatgggac aataacaacc aaggagctgg
     1741 ggacggtgat gcggtctctg gggcagaacc ccacagaagc agagctgcag gacatgatca
     1801 atgaagtaga tgccgacggt gacggcacaa tcgacttccc tgagttcctg acaatgatgg
     1861 caagaaaaat gaaatacaca gacagtgaag aagaaattag agaagcgttc cgtgtgtttg
     1921 ataaggatgg caatggctac atcagtgcag cagagcttcg ccacgtgatg acaaaccttg
     1981 gagagaagtt aacagatgaa gaggttgatg aaatgatcag ggaagcagac atcgatgggg
     2041 atggtcaggt aaactacgaa gagtttgtac aaatgatgac agcgaagtaa gcggccgcga
     2101 ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2161 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2221 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2281 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     2341 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     2401 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2461 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2521 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     2581 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     2641 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2701 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     2761 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     2821 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2881 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     2941 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3001 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3061 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3121 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3181 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3241 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     3301 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3361 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3421 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3481 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3541 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3601 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3661 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3721 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     3781 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     3841 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     3901 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     3961 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4021 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4081 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4141 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4201 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4261 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     4321 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     4381 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     4441 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     4501 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     4561 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4621 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4681 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4741 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4801 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4861 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4921 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4981 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5041 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5101 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5161 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5221 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5281 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5341 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5401 gataaccgta ttaccgccat gcat
//
