LOCUS       sequence-386858-        5224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of ultrafast protein calcium sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5224)
  AUTHORS   Zhang Y, Rozsa M, Bushey D, Zheng J, Reep D, Broussard GJ, Tsang A, 
            Tsegaye G, Patel R, Narayan S, Li JX, Zhang R, Ahrens MB, Turner GC,
            Wang SS-H, Svoboda K, Korff W, Schreiter ER, Hasseman JP, Kolb I, 
            Looger LL
  TITLE     jGCaMP8 Fast Genetically Encoded Calcium Indicators
  JOURNAL   Janelia Research Campus, Online Resource (2020) 
            doi:10.25378/janelia.13148243.
REFERENCE   2  (bases 1 to 5224)
  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: "Janelia 
            Research Campus, Online Resource (2020) 
            doi:10.25378/janelia.13148243."
COMMENT     Sequence Label: pGP-CMV-jGCaMP8f
FEATURES             Location/Qualifiers
     source          1..5224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        97..400
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        401..604
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     555..575
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             661..678
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     960..981
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1087..1108)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1348..1367)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2048..2169
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2085..2104)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2139..2158
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2176..2631)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2313..2334)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2525..2544
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2658..2762
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2764..3121
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2764..2955
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2787..2807)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2972..3107
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3156..3950
                     /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             3156..3950
                     /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(3210..3229)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3820..3839
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4138..4157)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4182..4229
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4558..5146
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5047..5066
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
       61 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      121 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      181 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      241 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      301 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      361 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      421 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      481 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      541 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      601 agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct cgccaccatg
      661 catcatcacc atcatcacac gcgtcgcaag aagaccttca aggaggtggc caccgccgtg
      721 aagatcatcg cccgcctgat gggcctgaag atcaacgtct atatcaaggc cgacaagcag
      781 aagaacggca tcaaggcgaa cttccacatc cgccacaaca tcgaggacgg cggcgtgcag
      841 ctcgcctacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
      901 aaccactacc tgagcgtgga gtccaaactt tcgaaagacc ccaacgagaa gcgcgatcac
      961 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1021 aagggcggta ccggagggag catggtgagc aagggcgagg agctgttcac cggggtggtg
     1081 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1141 ggtgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1201 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     1261 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1321 atccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1381 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1441 gacggcaaca tcctggggca caagctggag tacaacctgc cggaccaact gactgaagag
     1501 cagatcgcag aatttaaaga ggctttctcc ctatttgaca aggacgggga tgggacaata
     1561 acaaccaagg agctggggac ggtgatgcgg tctctcggac tgaaccccac agaagcagag
     1621 ctgcaggaca tgatcaatga agtagatgcc gacggtgacg gcacaatcga cttccctgag
     1681 ttcctgacaa tgatggcaag aaaaatgaaa tacagggaca cggaagaaga aattagagaa
     1741 gcgttcggtg tgtttgataa ggatggcaat ggctacatca gtgcagcaga gcttcgccac
     1801 gtgatgacaa accttggaga gaagttaaca gatgaagagg ttgatgaaat gatcagggaa
     1861 gcagacatcg atggggatgg tcaggtaaac tacgaagagt ttgtacaaat gatgacagcg
     1921 aagtaagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     1981 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2041 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2101 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2161 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     2221 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     2281 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2341 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2401 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     2461 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     2521 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     2581 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     2641 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2701 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2761 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2821 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     2881 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2941 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3001 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3061 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3121 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3181 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3241 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3301 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     3361 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3421 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3481 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3541 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3601 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3661 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     3721 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3781 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3841 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3901 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     3961 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4021 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4081 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     4141 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     4201 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     4261 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     4321 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     4381 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     4441 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4501 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4561 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4621 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4681 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4741 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4801 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4861 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4921 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4981 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5041 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5101 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5161 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5221 atcc
//
