LOCUS       sequence-366069-        5500 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of small FRET-based near-infrared genetically-encoded 
            calcium indicator in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5500)
  AUTHORS   Matlashov ME, Vera J, Kasatkina LA, Khodakhah K, Verkhusha VV
  TITLE     Design and Initial Characterization of a Small Near-Infrared 
            Fluorescent Calcium Indicator.
  JOURNAL   Front Cell Dev Biol. 2022 Jun 29;10:880107. doi: 
            10.3389/fcell.2022.880107. eCollection 2022.
  PUBMED    35846350
REFERENCE   2  (bases 1 to 5500)
  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 
            Dev Biol."; date: "2022-06-29"; volume: "10:880107. doi"; pages: " 
            10.3389/fcell.2022.880107. eCollection 2022"
COMMENT     Sequence Label: piGECInano-N1
FEATURES             Location/Qualifiers
     source          1..5500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        71..374
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        375..578
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     529..549
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      605..614
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2298..2419
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2335..2354)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2389..2408
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2426..2881)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2563..2584)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2775..2794
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2908..3012
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3014..3371
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3014..3205
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3037..3057)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3222..3357
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3406..4200
                     /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             3406..4200
                     /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(3460..3479)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4070..4089
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4388..4407)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4432..4479
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4808..5396
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5297..5316
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
       61 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      121 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      181 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      241 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      301 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      361 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      421 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      481 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      541 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
      601 gctagccacc atggccaatc tcgataaaat gctcaataca acagtgaccg aggtgcgcaa
      661 gttcctgcaa gcggacagag tgtgcgtgtt caagttcgag gaagattact ccggcaccgt
      721 ctcgcacgaa gccgtggacg acagatggat tagcatcctg aagacccagg tgcaggacag
      781 atacttcatg gaaaccagag gcgaggaata cgtccacggc agataccagg ccatcgccga
      841 catctacaca gccaatctgg tcgagtgcta cagagacctg ctgatcgagt ttcaggtgcg
      901 ggccattctg gctgtcccca tcctgcaagg caagaagctg tggggcctgc tggtggccca
      961 ccaactggcc ggccctcggg agtggcagac ctgggaaatc gacttcctga aacagcaagc
     1021 tgtcgtcatg ggaattgcta ttcaacagcc gccaagcgaa gaggaactga gcgagtgctt
     1081 cagaatcttc gacttcgacg gcaacggctt catcgacaga gaggagtttg gcgacatcat
     1141 cagactgacc ggcgagcagc tgaccgacga ggacgtggac gagatcttcg gcgactccga
     1201 caccgacaag aacggcagaa tcgatttcga cgagttcctg aagatggtgg aaaacgtgca
     1261 gccgacctgc gacgatgagc cgatccatat ccccggtgcc atccaaccgc atggactgct
     1321 gctcgccctc gccgccgaca tgacgatcgt tgccggcagc gacaaccttc ccgaactcac
     1381 cggactggcg atcggcgccc tgatcggccg ctctgcggcc gatgtcttcg actcggagac
     1441 gcacaaccgt ctgacgatcg ccttggccga gcccggggcg gccgtcggag caccgatcac
     1501 tgtcggcttc acgatgcgaa aggacgcagg cttcatcggc tcctggcatc gccatgatca
     1561 gctcatcttc ctggagctgg agcctcccca gcgggacgtc gccgagccgc aggcgttctt
     1621 ccgccgcacc aacagcgcca tccgccgcct gcaggccgcc gaaaccttgg aaagcgcctg
     1681 cgccgccgcg gcgcaagagg tgcggaagat taccggcttc gatcgggtga tgatctatcg
     1741 cttcgcctcc gacttcagcg ggtccgtgat cgcagaggat cggtgcgccg aggtcgagtc
     1801 aaaactaggc ctgcactatc ctgcctcatt catcccggcg caggcccgtc ggctctatac
     1861 catcaacccg gtacggatca ttcccgacat caattatcgg ccggtgccgg tcaccccaga
     1921 cctcaatccg gtcaccgggc ggccgattga tcttagcttc gccatcctgc gcagcgtctc
     1981 gcccaaccat ctggagttca tgcgcaacat aggcatgcac ggcacgatgt cgatctcgat
     2041 tttgcgcggc gagcgactgt ggggattgat cgtttgccat caccgaacgc cgtactacgt
     2101 cgatctcgat ggccgccaag cctgcaagag ggtcgccgag aggctggcca ctcagatcgg
     2161 cgtgatggaa gagtgagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     2221 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     2281 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     2341 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2401 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     2461 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     2521 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     2581 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2641 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2701 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2761 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2821 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2881 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2941 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3001 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     3061 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3121 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3181 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3241 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3301 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3361 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3421 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3481 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3541 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     3601 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3661 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3721 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3781 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3841 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3901 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     3961 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4021 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4081 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4141 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4201 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4261 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4321 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     4381 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     4441 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     4501 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     4561 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     4621 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     4681 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4741 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4801 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4861 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4921 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4981 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5041 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5101 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5161 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5221 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5281 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5341 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5401 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5461 ttcctgcgtt atcccctgat tctgtggata accgtattac
//
