LOCUS       sequence-372990-        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-jGCaMP8s
FEATURES             Location/Qualifiers
     source          1..5224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        127..318
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     275..295
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(320..424)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      451..906
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(538..557)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     748..769
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    913..1034
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(924..943)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     978..997
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1715..1734
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1974..1995
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2101..2122)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(2404..2421)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(2478..2681)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2507..2527)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2682..2985
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3160..3748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3240..3259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4077..4124
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4149..4168
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4356..5150)
                     /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             complement(4356..5150)
                     /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(4467..4486)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5077..5096
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5185..5224,1..318))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5199..5224,1..110)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt
       61 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt
      121 aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct
      181 ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata
      241 cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac attccacagc
      301 tggttctttc cgcctcagga ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
      361 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
      421 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
      481 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
      541 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
      601 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
      661 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
      721 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
      781 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
      841 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
      901 caatttacgc cttaagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
      961 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     1021 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     1081 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc
     1141 tagagtcgcg gccgcttact tcgctgtcat catttgtaca aactcttcgt agtttacctg
     1201 accatcccca tcgatgtctg cttccctgat catttcatca acctcttcat ctgttaactt
     1261 ctctccaagg tttgtcatca cgtggcgaag ctctgctgca ctgatgtagc cattgccatc
     1321 cttatcaaac acaccgaacg cttctctaat ttcttcttcc gtgtccctgt atttcatttt
     1381 tcttgccatc attgtcagga actcagggaa gtcgattgtg ccgtcaccgt cggcatctac
     1441 ttcattgatc atgtcctgca gctctgcttc tgtggggttg tgtccgagag accgcatcac
     1501 cgtccccagc tccttggttg ttattgtccc atccccgtcc ttgtcaaata gggagaaagc
     1561 ctctttgtat tctgcgatct gctcttcagt cagttggtcc ggcaggttgt actccagctt
     1621 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
     1681 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa
     1741 gaagatggtg cgctcctgga tgtagccttc gggcatggcg gacttgaaga agtcgtgctg
     1801 cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg tggtcacgag
     1861 ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc
     1921 gtaggtggca tcgccctcac cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc
     1981 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat
     2041 gctccctccg gtaccgccct tgtacagctc gtccatgccg agagtgatcc cggcggcggt
     2101 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttcg aaagtttgga
     2161 ctccacgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
     2221 ctgctggtag tggtaggcga gctgcacgcc gccgtcctcg atgttgtggc ggatgtggaa
     2281 gttcgccttg atgccgttct tctgcttgtc ggccttgata tagacgttga tcttcaggcc
     2341 catcagcatg gcgatgatct tcacggcggt ggccacctcc ttgaaggtct tcttgcgacg
     2401 cgtgtgatga tggtgatgat gcatggtggc gagatctgag tccggtagcg ctagcggatc
     2461 tgacggttca ctaaaccagc tctgcttata tagacctccc accgtacacg cctaccgccc
     2521 atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc
     2581 caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac
     2641 cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact
     2701 aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc
     2761 caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact
     2821 tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa
     2881 gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg
     2941 ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat
     3001 gggctatgaa ctaatgaccc cgtaattgat tactattaat aactaatgca tggcggtaat
     3061 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3121 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3181 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3241 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3301 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3361 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3421 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3481 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3541 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3601 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3661 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3721 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3781 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3841 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3901 gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc
     3961 acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg
     4021 gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc gtttatgaac
     4081 aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag cgcgggttcc
     4141 ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg cgaagaactc
     4201 cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa
     4261 gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt
     4321 cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga
     4381 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
     4441 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
     4501 cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
     4561 atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg
     4621 ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
     4681 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
     4741 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
     4801 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
     4861 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg
     4921 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc
     4981 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
     5041 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
     5101 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat
     5161 cctcatcctg tctcttgatc gatctttgca aaagcctagg cctccaaaaa agcctcctca
     5221 ctac
//
