LOCUS       sequence-310688-        6330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cytoplasmic expression of a near-infrared genetically encoded 
            calcium indicator iGECI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6330)
  AUTHORS   Shemetov AA, Monakhov MV, Zhang Q, Canton-Josh JE, Kumar M, Chen M, 
            Matlashov ME, Li X, Yang W, Nie L, Shcherbakova DM, Kozorovitskiy Y,
            Yao J, Ji N, Verkhusha VV
  TITLE     A near-infrared genetically encoded calcium indicator for in vivo 
            imaging.
  JOURNAL   Nat Biotechnol. 2021 Mar;39(3):368-377. doi: 
            10.1038/s41587-020-0710-1. Epub 2020 Oct 26.
  PUBMED    33106681
REFERENCE   2  (bases 1 to 6330)
  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: 
            "10.1038/s41587-020-0710-1"; journalName: "Nat Biotechnol"; date: 
            "2021-03"; volume: "39"; issue: "3"; pages: "368-377"
COMMENT     Sequence Label: piGECI-N1
FEATURES             Location/Qualifiers
     source          1..6330
                     /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      597..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2001..2078
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    3118..3239
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3155..3174)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3209..3228
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3246..3701)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3383..3404)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3595..3614
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3728..3832
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3834..4191
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3834..4025
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3857..3877)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4042..4177
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4226..5020
                     /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             4226..5020
                     /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(4280..4299)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4890..4909
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5208..5227)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5252..5299
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5628..6216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6117..6136
                     /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 gctagcgcca
      601 ccatggcgga aggctccgtc gccaggcagc ctgacctctt gacctgcgaa catgaagaga
      661 tccacctcgc cggctcgatc cagccgcatg gcgcgcttct ggtcgtcagc gaacatgatc
      721 atcgcgtcat ccaggccagc gccaacgccg cggaatttct gaatctcgga agcgtactcg
      781 gcgttccgct cgccgagatc gacggcgatc tgttgatcaa gatcctgccg catctcgatc
      841 ccaccgccga aggcatgccg gtcgcggtgc gctgccggat cggcaatccc tctacggagt
      901 actgcggtct gatgcatcgg cctccggaag gcgggctgat catcgaactc gaacgtgccg
      961 gcccgtcgat cgatctgtca ggcacgctgg cgccggcgct ggagcggatc cgcacggcgg
     1021 gttcactgcg cgcgctgtgc gatgacaccg tgctgctgtt tcagcagtgc accggctacg
     1081 accgggtgat ggtgtatcgt ttcgatgagc aaggccacgg cctggtattc tccgagtgcc
     1141 atgtgcctgg gctcgaatcc tatttcggca accgctatcc gtcgtcgact gtcccgcaga
     1201 tggcgcggca gctgtacgtg cggcagcgcg tccgcgtgct ggtcgacgtc acctatcagc
     1261 cggtgccgct ggagccgcgg ctgtcgccgc tgaccgggcg cgatctcgac atgtcgggct
     1321 gcttcctgcg ctcgatgtcg ccgtgccatc tgcagttcct gaaggacatg ggcgtgcgcg
     1381 ccaccctggc ggtgtcgctg gtggtcggcg gcaagctgtg gggcctggtt gtctgtcacc
     1441 attatctgcc gcgcttcatc cgtttcgagc tgcgggcgat ctgcaaacgg ctcgccgaaa
     1501 ggatcgcgac gcggatcacc gcgctttaca cccatgacca actgacagaa gagcagattg
     1561 cagagttcaa agaagccttc tcattattcg acaaggatgg ggacggcacc atcaccacaa
     1621 aggaacttgg caccgttatg aggtcgcttg gacaaaaccc aacggaagca gaattgcagg
     1681 atatgatcaa tgaagtcgat gctgatggca atggaacgat ttactttcct gaatttctta
     1741 ctatgatggc tagaaaaatg aaggacacag acagcgaaga ggaaatccga gaagcattcc
     1801 gtgtttttga caaggatggg aacggctaca tcagcgctgc tgaattacgt cacgtcatga
     1861 caaacctcgg ggagaagtta acagatgaag aagttgatga aatgataagg gaagcagata
     1921 tcgatggtga tggccaagta aactatgaag agtttgtaca aatgatgaca gcaaaggggg
     1981 ggggtggcgg aggtggctcc aagaggcgct ggaagaaaaa cttcattgcc gtcagcgctg
     2041 ccaaccggtt caagaagata tccagctccg gggcactggt agtaacctgc gacgatgagc
     2101 cgatccatat ccccggtgcc atccaaccgc atggactgct gctcgccctc gccgccgaca
     2161 tgacgatcgt tgccggcagc gacaaccttc ccgaactcac cggactggcg atcggcgccc
     2221 tgatcggccg ctctgcggcc gatgtcttcg actcggagac gcacaaccgt ctgacgatcg
     2281 ccttggccga gcccggggcg gccgtcggag caccgatcac tgtcggcttc acgatgcgaa
     2341 aggacgcagg cttcatcggc tcctggcatc gccatgatca gctcatcttc ctcgagctcg
     2401 agcctcccca gcgggacgtc gccgagccgc aggcgttctt ccgccgcacc aacagcgcca
     2461 tccgccgcct gcaggccgcc gaaaccttgg aaagcgcctg cgccgccgcg gcgcaagagg
     2521 tgcggaagat taccggcttc gatcgggtga tgatctatcg cttcgcctcc gacttcagcg
     2581 ggtccgtgat cgcagaggat cggtgcgccg aggtcgagtc aaaactaggc ctgcactatc
     2641 ctgcctcatt catcccggcg caggcccgtc ggctctatac catcaacccg gtacggatca
     2701 ttcccgatat caattatcgg ccggtgccgg tcaccccaga cctcaatccg gtcaccgggc
     2761 ggccgattga tcttagcttc gccatcctgc gcagcgtctc gcccaaccat ctggagttca
     2821 tgcgcaacat aggcatgcac ggcacgatgt cgatctcgat tttgcgcggc gagcgactgt
     2881 ggggattgat cgtttgccat caccgaacgc cgtactacgt cgatctcgat ggccgccaag
     2941 cctgcaagag gctcgccgag aggctggcca ctcagatcgg cgtgatggaa gagtgagcgg
     3001 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     3061 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     3121 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3181 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     3241 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3301 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3361 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3421 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3481 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3541 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3601 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3661 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     3721 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3781 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     3841 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3901 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3961 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4021 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4081 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4141 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     4201 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4261 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4321 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4381 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     4441 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4501 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4561 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4621 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4681 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4741 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4801 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4861 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4921 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4981 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5041 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5101 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5161 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     5221 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     5281 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     5341 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     5401 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     5461 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5521 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5581 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5641 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5701 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5761 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5821 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5881 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5941 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6001 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6061 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6121 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6181 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6241 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6301 tctgtggata accgtattac cgccatgcat
//
