LOCUS       sequence-350822-        6821 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Red intensiometric genetically encoded Ca2+-indicators for optical 
            imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6821)
  AUTHORS   Wu J, Liu L, Matsuda T, Zhao Y, Rebane A, Drobizhev M, Chang YF, 
            Araki S, Arai Y, March K, Hughes TE, Sagou K, Miyata T, Nagai T, Li 
            WH, Campbell RE
  TITLE     Improved Orange and Red Ca Indicators and Photophysical 
            Considerations for Optogenetic Applications.
  JOURNAL   ACS Chem Neurosci. 2013 Mar 19.
  PUBMED    23452507
REFERENCE   2  (bases 1 to 6821)
  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: "ACS Chem 
            Neurosci. 2013 Mar 19."
COMMENT     Sequence Label: CMV-mito-R-GECO1
FEATURES             Location/Qualifiers
     source          1..6821
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(8..28)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        13..342
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      193..328
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     255..274
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             409..1203
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(463..482)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1073..1092
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1377..1498
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1414..1433)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1468..1487
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1547..1563)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1547..1563)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1560..1582)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1571..1587
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1595..1601,1602..1619,1620..1625))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1640..1661
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1778..1795)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1949..2537)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2029..2048)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2708..3499,3500..3568))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3331..3350
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3569..3673)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3748..3767)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3939..4318
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4319..4522
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4473..4493
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4567..4586
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4567..4585
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4632..4706
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             4728..4802
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             4851..4907
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     5603..5622
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             5646..6089
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTFDFPEFLTMMARKMNDTDSEEEIREAFRVFDKDGNGYIGAAEL
                     RHVMTDLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
     primer_bind     complement(6116..6133)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6122..6346
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6392..6820
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6479..6498)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6689..6710
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
       61 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
      121 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
      181 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
      241 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
      301 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
      361 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
      421 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
      481 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
      541 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
      601 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
      661 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
      721 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
      781 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
      841 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
      901 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
      961 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     1021 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     1081 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     1141 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     1201 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     1261 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     1321 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     1381 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     1441 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     1501 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     1561 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     1621 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     1681 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     1741 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     1801 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     1861 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     1921 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1981 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2041 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2101 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2161 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2221 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2281 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2341 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2401 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2461 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2521 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2581 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2641 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2701 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2761 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     2821 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     2881 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     2941 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3001 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3061 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3121 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3181 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3241 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3301 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3361 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3421 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3481 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3541 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3601 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3661 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     3721 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3781 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     3841 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     3901 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
     3961 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     4021 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     4081 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     4141 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     4201 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     4261 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     4321 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     4381 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     4441 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     4501 ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt actggcttat
     4561 cgaaattaat acgactcact atagggagac ccaagctggc tagcgtttaa acgggccctc
     4621 tagactcgag catgtctgtt ctgactcctc tgctgctccg gggtctcaca ggttccgcaa
     4681 gaagactccc cgtgcctagg gccaaaattc attcactggg ggaccccatg agcgtgctca
     4741 ccccactcct gctgcggggg ctgaccggca gcgctaggcg gctgccagtc cccagggcca
     4801 agatccacag tctcggcgat cccaaggatc caaccatggt cgactcttca cgtcgtaagt
     4861 ggaataaggc aggtcacgca gtcagagcta taggtcggct gagctcaccc gtggtttccg
     4921 agcggatgta ccccgaggac ggcgccctga agagcgagat caagaagggg ctgaggctga
     4981 aggacggcgg ccactacgcc gccgaggtca agaccaccta caaggccaag aagcccgtgc
     5041 agctgcccgg cgcctacatc gtcgacatca agttggacat cgtgtcccac aacgaggact
     5101 acaccatcgt ggaacagtgc gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     5161 agctgtacaa gggaggtaca ggcgggagtc tggtgagcaa gggcgaggag gataacatgg
     5221 ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg
     5281 agttcgagat cgagggcgag ggcgagggcc gcccctacga ggcctttcag accgctaagc
     5341 tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca
     5401 tgtacggctc caaggcctac attaagcacc cagccgacat ccccgactac ttcaagctgt
     5461 ccttccccga gggcttcagg tgggagcgcg tgatgaactt cgaggacggc ggcattattc
     5521 acgttaacca ggactcctcc ctgcaggacg gcgtattcat ctacaaggtg aagctgcgcg
     5581 gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcta
     5641 cgcgtgacca actgactgaa gagcagatcg cagaatttaa agaggctttc tccctatttg
     5701 acaaggacgg ggatgggacg ataacaacca aggagctggg gacggtgatg cggtctctgg
     5761 ggcagaaccc cacagaagca gagctgcagg acatgatcaa tgaagtagat gccgacggtg
     5821 acggcacatt cgacttccct gagttcctga cgatgatggc aagaaaaatg aatgacacag
     5881 acagtgaaga ggaaattaga gaagcgttcc gcgtgtttga taaggacggc aatggctaca
     5941 tcggcgcagc agagcttcgc cacgtgatga cagaccttgg agagaagtta acagatgagg
     6001 aggttgatga aatgatcagg gtagcagaca tcgatgggga tggtcaggta aactacgaag
     6061 agtttgtaca aatgatgaca gcgaagtaga agcttaagtt taaaccgctg atcagcctcg
     6121 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     6181 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     6241 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     6301 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     6361 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     6421 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     6481 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     6541 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     6601 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     6661 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     6721 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     6781 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt a
//
