LOCUS       sequence-315685-        6378 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a near-infrared genetically encoded calcium indicator 
            iGECI with a nuclear export signal sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6378)
  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 6378)
  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-NES-N1
FEATURES             Location/Qualifiers
     source          1..6378
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      233..242
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1637..1714
                     /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"
     CDS             2648..2674
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     polyA_signal    2802..2923
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2839..2858)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2893..2912
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2930..3385)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3067..3088)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3279..3298
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3412..3516
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3518..3875
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3518..3709
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3541..3561)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3726..3861
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3910..4704
                     /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             3910..4704
                     /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(3964..3983)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4574..4593
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4892..4911)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4936..4983
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5312..5900
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5801..5820
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6075..6378
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgccaccat
      241 ggcggaaggc tccgtcgcca ggcagcctga cctcttgacc tgcgaacatg aagagatcca
      301 cctcgccggc tcgatccagc cgcatggcgc gcttctggtc gtcagcgaac atgatcatcg
      361 cgtcatccag gccagcgcca acgccgcgga atttctgaat ctcggaagcg tactcggcgt
      421 tccgctcgcc gagatcgacg gcgatctgtt gatcaagatc ctgccgcatc tcgatcccac
      481 cgccgaaggc atgccggtcg cggtgcgctg ccggatcggc aatccctcta cggagtactg
      541 cggtctgatg catcggcctc cggaaggcgg gctgatcatc gaactcgaac gtgccggccc
      601 gtcgatcgat ctgtcaggca cgctggcgcc ggcgctggag cggatccgca cggcgggttc
      661 actgcgcgcg ctgtgcgatg acaccgtgct gctgtttcag cagtgcaccg gctacgaccg
      721 ggtgatggtg tatcgtttcg atgagcaagg ccacggcctg gtattctccg agtgccatgt
      781 gcctgggctc gaatcctatt tcggcaaccg ctatccgtcg tcgactgtcc cgcagatggc
      841 gcggcagctg tacgtgcggc agcgcgtccg cgtgctggtc gacgtcacct atcagccggt
      901 gccgctggag ccgcggctgt cgccgctgac cgggcgcgat ctcgacatgt cgggctgctt
      961 cctgcgctcg atgtcgccgt gccatctgca gttcctgaag gacatgggcg tgcgcgccac
     1021 cctggcggtg tcgctggtgg tcggcggcaa gctgtggggc ctggttgtct gtcaccatta
     1081 tctgccgcgc ttcatccgtt tcgagctgcg ggcgatctgc aaacggctcg ccgaaaggat
     1141 cgcgacgcgg atcaccgcgc tttacaccca tgaccaactg acagaagagc agattgcaga
     1201 gttcaaagaa gccttctcat tattcgacaa ggatggggac ggcaccatca ccacaaagga
     1261 acttggcacc gttatgaggt cgcttggaca aaacccaacg gaagcagaat tgcaggatat
     1321 gatcaatgaa gtcgatgctg atggcaatgg aacgatttac tttcctgaat ttcttactat
     1381 gatggctaga aaaatgaagg acacagacag cgaagaggaa atccgagaag cattccgtgt
     1441 ttttgacaag gatgggaacg gctacatcag cgctgctgaa ttacgtcacg tcatgacaaa
     1501 cctcggggag aagttaacag atgaagaagt tgatgaaatg ataagggaag cagatatcga
     1561 tggtgatggc caagtaaact atgaagagtt tgtacaaatg atgacagcaa aggggggggg
     1621 tggcggaggt ggctccaaga ggcgctggaa gaaaaacttc attgccgtca gcgctgccaa
     1681 ccggttcaag aagatatcca gctccggggc actggtagta acctgcgacg atgagccgat
     1741 ccatatcccc ggtgccatcc aaccgcatgg actgctgctc gccctcgccg ccgacatgac
     1801 gatcgttgcc ggcagcgaca accttcccga actcaccgga ctggcgatcg gcgccctgat
     1861 cggccgctct gcggccgatg tcttcgactc ggagacgcac aaccgtctga cgatcgcctt
     1921 ggccgagccc ggggcggccg tcggagcacc gatcactgtc ggcttcacga tgcgaaagga
     1981 cgcaggcttc atcggctcct ggcatcgcca tgatcagctc atcttcctcg agctcgagcc
     2041 tccccagcgg gacgtcgccg agccgcaggc gttcttccgc cgcaccaaca gcgccatccg
     2101 ccgcctgcag gccgccgaaa ccttggaaag cgcctgcgcc gccgcggcgc aagaggtgcg
     2161 gaagattacc ggcttcgatc gggtgatgat ctatcgcttc gcctccgact tcagcgggtc
     2221 cgtgatcgca gaggatcggt gcgccgaggt cgagtcaaaa ctaggcctgc actatcctgc
     2281 ctcattcatc ccggcgcagg cccgtcggct ctataccatc aacccggtac ggatcattcc
     2341 cgatatcaat tatcggccgg tgccggtcac cccagacctc aatccggtca ccgggcggcc
     2401 gattgatctt agcttcgcca tcctgcgcag cgtctcgccc aaccatctgg agttcatgcg
     2461 caacataggc atgcacggca cgatgtcgat ctcgattttg cgcggcgagc gactgtgggg
     2521 attgatcgtt tgccatcacc gaacgccgta ctacgtcgat ctcgatggcc gccaagcctg
     2581 caagaggctc gccgagaggc tggccactca gatcggcgtg atggaagagg gaggtggaac
     2641 cctgcagctg cctcctctgg aacgcctgac tctggattga gcggccgcga ctctagatca
     2701 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     2761 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     2821 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2881 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     2941 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3001 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3061 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3121 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3181 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3241 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3301 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     3361 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3421 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3481 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     3541 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3601 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3661 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3721 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3781 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3841 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     3901 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3961 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4021 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4081 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4141 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4201 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4261 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4321 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4381 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     4441 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4501 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4561 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4621 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4681 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4741 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4801 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4861 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     4921 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     4981 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     5041 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     5101 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     5161 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5221 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5281 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5341 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5401 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5461 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5521 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5581 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5641 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5701 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5761 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5821 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5881 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5941 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6001 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     6061 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     6121 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     6181 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     6241 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     6301 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     6361 gtcatcgcta ttaccatg
//
