LOCUS       sequence-416763-        5224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for calcium imaging in mammalian cells - 
            jGCaMP8f with L27A mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5224)
  AUTHORS   Tran O, Hughes HJ, Carter T, Torok K
  TITLE     Development and characterization of novel jGCaMP8f calcium sensor 
            variants with improved kinetics and fluorescence response range.
  JOURNAL   Front Cell Neurosci. 2023 May 18;17:1155406. doi: 
            10.3389/fncel.2023.1155406. eCollection 2023.
  PUBMED    37275778
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: "Front Cell 
            Neurosci."; date: "2023-05-18"; volume: "17:1155406. doi"; pages: " 
            10.3389/fncel.2023.1155406. eCollection 2023"
COMMENT     Sequence Label: pGP-CMV-jGCaMP8f L27A
FEATURES             Location/Qualifiers
     source          1..5224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     498..517
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(816..835)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    860..907
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1236..1824
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1725..1744
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1999..2302
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2303..2506
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2457..2477
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2563..2580
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     2862..2883
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2989..3010)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3250..3269)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3950..4071
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3987..4006)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4041..4060
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4078..4533)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4215..4236)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4427..4446
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4560..4664
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4666..5023
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4666..4857
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4689..4709)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4874..5009
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(5058..5224,1..628)
                     /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             join(5058..5224,1..628)
                     /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(5112..5131)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
       61 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
      121 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
      181 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
      241 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
      301 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
      361 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
      421 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
      481 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
      541 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
      601 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
      661 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
      721 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
      781 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
      841 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
      901 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
      961 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     1021 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     1081 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1141 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     1201 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1261 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1321 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1381 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1441 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1501 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     1561 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     1621 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     1681 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     1741 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     1801 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     1861 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     1921 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
     1981 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     2041 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     2101 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     2161 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     2221 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     2281 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     2341 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     2401 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     2461 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
     2521 tcagatccgc tagcgctacc ggactcagat ctcgccacca tgcatcatca ccatcatcac
     2581 acgcgtcgca agaagacctt caaggaggtg gccaccgccg tgaagatcat cgcccgcgca
     2641 atgggcctga agatcaacgt ctatatcaag gccgacaagc agaagaacgg catcaaggcg
     2701 aacttccaca tccgccacaa catcgaggac ggcggcgtgc agctcgccta ccactaccag
     2761 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcgtg
     2821 gagtccaaac tttcgaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2881 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggcgg taccggaggg
     2941 agcatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     3001 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggtgaggg cgatgccacc
     3061 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     3121 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     3181 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acatccagga gcgcaccatc
     3241 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     3301 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     3361 cacaagctgg agtacaacct gccggaccaa ctgactgaag agcagatcgc agaatttaaa
     3421 gaggctttct ccctatttga caaggacggg gatgggacaa taacaaccaa ggagctgggg
     3481 acggtgatgc ggtctctcgg actgaacccc acagaagcag agctgcagga catgatcaat
     3541 gaagtagatg ccgacggtga cggcacaatc gacttccctg agttcctgac aatgatggca
     3601 agaaaaatga aatacaggga cacggaagaa gaaattagag aagcgttcgg tgtgtttgat
     3661 aaggatggca atggctacat cagtgcagca gagcttcgcc acgtgatgac aaaccttgga
     3721 gagaagttaa cagatgaaga ggttgatgaa atgatcaggg aagcagacat cgatggggat
     3781 ggtcaggtaa actacgaaga gtttgtacaa atgatgacag cgaagtaagc ggccgcgact
     3841 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     3901 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     3961 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4021 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     4081 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     4141 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     4201 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     4261 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     4321 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     4381 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     4441 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     4501 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     4561 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4621 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     4681 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4741 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4801 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4861 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4921 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4981 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     5041 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     5101 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     5161 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     5221 gtgc
//
