LOCUS       sequence-315693-        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        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      537..546
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1941..2018
                     /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    3058..3179
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3095..3114)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3149..3168
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3186..3641)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3323..3344)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3535..3554
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3668..3772
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3774..4131
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3774..3965
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3797..3817)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3982..4117
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4166..4960
                     /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             4166..4960
                     /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(4220..4239)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4830..4849
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5148..5167)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5192..5239
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5568..6156
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6057..6076
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcca
      541 ccatggcgga aggctccgtc gccaggcagc ctgacctctt gacctgcgaa catgaagaga
      601 tccacctcgc cggctcgatc cagccgcatg gcgcgcttct ggtcgtcagc gaacatgatc
      661 atcgcgtcat ccaggccagc gccaacgccg cggaatttct gaatctcgga agcgtactcg
      721 gcgttccgct cgccgagatc gacggcgatc tgttgatcaa gatcctgccg catctcgatc
      781 ccaccgccga aggcatgccg gtcgcggtgc gctgccggat cggcaatccc tctacggagt
      841 actgcggtct gatgcatcgg cctccggaag gcgggctgat catcgaactc gaacgtgccg
      901 gcccgtcgat cgatctgtca ggcacgctgg cgccggcgct ggagcggatc cgcacggcgg
      961 gttcactgcg cgcgctgtgc gatgacaccg tgctgctgtt tcagcagtgc accggctacg
     1021 accgggtgat ggtgtatcgt ttcgatgagc aaggccacgg cctggtattc tccgagtgcc
     1081 atgtgcctgg gctcgaatcc tatttcggca accgctatcc gtcgtcgact gtcccgcaga
     1141 tggcgcggca gctgtacgtg cggcagcgcg tccgcgtgct ggtcgacgtc acctatcagc
     1201 cggtgccgct ggagccgcgg ctgtcgccgc tgaccgggcg cgatctcgac atgtcgggct
     1261 gcttcctgcg ctcgatgtcg ccgtgccatc tgcagttcct gaaggacatg ggcgtgcgcg
     1321 ccaccctggc ggtgtcgctg gtggtcggcg gcaagctgtg gggcctggtt gtctgtcacc
     1381 attatctgcc gcgcttcatc cgtttcgagc tgcgggcgat ctgcaaacgg ctcgccgaaa
     1441 ggatcgcgac gcggatcacc gcgctttaca cccatgacca actgacagaa gagcagattg
     1501 cagagttcaa agaagccttc tcattattcg acaaggatgg ggacggcacc atcaccacaa
     1561 aggaacttgg caccgttatg aggtcgcttg gacaaaaccc aacggaagca gaattgcagg
     1621 atatgatcaa tgaagtcgat gctgatggca atggaacgat ttactttcct gaatttctta
     1681 ctatgatggc tagaaaaatg aaggacacag acagcgaaga ggaaatccga gaagcattcc
     1741 gtgtttttga caaggatggg aacggctaca tcagcgctgc tgaattacgt cacgtcatga
     1801 caaacctcgg ggagaagtta acagatgaag aagttgatga aatgataagg gaagcagata
     1861 tcgatggtga tggccaagta aactatgaag agtttgtaca aatgatgaca gcaaaggggg
     1921 ggggtggcgg aggtggctcc aagaggcgct ggaagaaaaa cttcattgcc gtcagcgctg
     1981 ccaaccggtt caagaagata tccagctccg gggcactggt agtaacctgc gacgatgagc
     2041 cgatccatat ccccggtgcc atccaaccgc atggactgct gctcgccctc gccgccgaca
     2101 tgacgatcgt tgccggcagc gacaaccttc ccgaactcac cggactggcg atcggcgccc
     2161 tgatcggccg ctctgcggcc gatgtcttcg actcggagac gcacaaccgt ctgacgatcg
     2221 ccttggccga gcccggggcg gccgtcggag caccgatcac tgtcggcttc acgatgcgaa
     2281 aggacgcagg cttcatcggc tcctggcatc gccatgatca gctcatcttc ctcgagctcg
     2341 agcctcccca gcgggacgtc gccgagccgc aggcgttctt ccgccgcacc aacagcgcca
     2401 tccgccgcct gcaggccgcc gaaaccttgg aaagcgcctg cgccgccgcg gcgcaagagg
     2461 tgcggaagat taccggcttc gatcgggtga tgatctatcg cttcgcctcc gacttcagcg
     2521 ggtccgtgat cgcagaggat cggtgcgccg aggtcgagtc aaaactaggc ctgcactatc
     2581 ctgcctcatt catcccggcg caggcccgtc ggctctatac catcaacccg gtacggatca
     2641 ttcccgatat caattatcgg ccggtgccgg tcaccccaga cctcaatccg gtcaccgggc
     2701 ggccgattga tcttagcttc gccatcctgc gcagcgtctc gcccaaccat ctggagttca
     2761 tgcgcaacat aggcatgcac ggcacgatgt cgatctcgat tttgcgcggc gagcgactgt
     2821 ggggattgat cgtttgccat caccgaacgc cgtactacgt cgatctcgat ggccgccaag
     2881 cctgcaagag gctcgccgag aggctggcca ctcagatcgg cgtgatggaa gagtgagcgg
     2941 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     3001 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     3061 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3121 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     3181 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3241 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3301 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3361 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3421 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3481 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3541 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3601 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     3661 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3721 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     3781 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3841 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3901 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3961 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4021 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4081 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     4141 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4201 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4261 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4321 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     4381 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4441 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4501 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4561 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4621 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4681 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4741 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4801 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4861 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4921 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4981 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5041 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5101 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     5161 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     5221 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     5281 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     5341 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     5401 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5461 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5521 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5581 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5641 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5701 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5761 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5821 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5881 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5941 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6001 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6061 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6121 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6181 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6241 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
     6301 attagttcat agcccatata tggagttccg
//
