LOCUS CMV-mito-R-GECO1 6821 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Red intensiometric genetically encoded Ca2+-indicators for optical
imaging.
ACCESSION .
VERSION .
KEYWORDS CMV-mito-R-GECO1
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 May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6821
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 26..405
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 406..609
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 560..580
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 654..673
/label=T7
/note="T7 promoter, forward primer"
promoter 654..672
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 719..793
/codon_start=1
/product="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
/label=COX8 presequence
/translation="MSVLTPLLLRGLTGSARRLPVPRAK"
CDS 815..889
/codon_start=1
/product="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
/label=COX8 presequence
/translation="MSVLTPLLLRGLTGSARRLPVPRAK"
CDS 938..994
/codon_start=1
/product="calmodulin-binding peptide from avian smooth
muscle myosin light chain kinase"
/label=calmodulin-binding peptide
/translation="RRKWNKAGHAVRAIGRLSS"
primer_bind 1690..1709
/label=mCherry-F
/note="mCherry, forward primer"
CDS 1733..2176
/codon_start=1
/product="calcium-binding protein"
/label=calmodulin
/translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
ELQDMINEVDADGDGTFDFPEFLTMMARKMNDTDSEEEIREAFRVFDKDGNGYIGAAEL
RHVMTDLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
primer_bind complement(2203..2220)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2209..2433
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2479..2907
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2566..2585)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2776..2797
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2916..2936)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2921..3250
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3101..3236
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3163..3182
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3317..4111
/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(3371..3390)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3981..4000
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4285..4406
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4322..4341)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4376..4395
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4455..4471)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4455..4471)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4468..4490)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4479..4495
/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(4503..4533)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4548..4569
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4686..4703)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4857..5445)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4937..4956)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5616..6476)
/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 6239..6258
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6477..6581)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6656..6675)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
61 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
121 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
181 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
241 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
301 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
361 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
421 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
481 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
541 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
601 agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg
661 actcactata gggagaccca agctggctag cgtttaaacg ggccctctag actcgagcat
721 gtctgttctg actcctctgc tgctccgggg tctcacaggt tccgcaagaa gactccccgt
781 gcctagggcc aaaattcatt cactggggga ccccatgagc gtgctcaccc cactcctgct
841 gcgggggctg accggcagcg ctaggcggct gccagtcccc agggccaaga tccacagtct
901 cggcgatccc aaggatccaa ccatggtcga ctcttcacgt cgtaagtgga ataaggcagg
961 tcacgcagtc agagctatag gtcggctgag ctcacccgtg gtttccgagc ggatgtaccc
1021 cgaggacggc gccctgaaga gcgagatcaa gaaggggctg aggctgaagg acggcggcca
1081 ctacgccgcc gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
1141 ctacatcgtc gacatcaagt tggacatcgt gtcccacaac gaggactaca ccatcgtgga
1201 acagtgcgaa cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaaggg
1261 aggtacaggc gggagtctgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
1321 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
1381 gggcgagggc gagggccgcc cctacgaggc ctttcagacc gctaagctga aggtgaccaa
1441 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
1501 ggcctacatt aagcacccag ccgacatccc cgactacttc aagctgtcct tccccgaggg
1561 cttcaggtgg gagcgcgtga tgaacttcga ggacggcggc attattcacg ttaaccagga
1621 ctcctccctg caggacggcg tattcatcta caaggtgaag ctgcgcggca ccaacttccc
1681 ccccgacggc cccgtaatgc agaagaagac catgggctgg gaggctacgc gtgaccaact
1741 gactgaagag cagatcgcag aatttaaaga ggctttctcc ctatttgaca aggacgggga
1801 tgggacgata acaaccaagg agctggggac ggtgatgcgg tctctggggc agaaccccac
1861 agaagcagag ctgcaggaca tgatcaatga agtagatgcc gacggtgacg gcacattcga
1921 cttccctgag ttcctgacga tgatggcaag aaaaatgaat gacacagaca gtgaagagga
1981 aattagagaa gcgttccgcg tgtttgataa ggacggcaat ggctacatcg gcgcagcaga
2041 gcttcgccac gtgatgacag accttggaga gaagttaaca gatgaggagg ttgatgaaat
2101 gatcagggta gcagacatcg atggggatgg tcaggtaaac tacgaagagt ttgtacaaat
2161 gatgacagcg aagtagaagc ttaagtttaa accgctgatc agcctcgact gtgccttcta
2221 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
2281 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
2341 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
2401 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
2461 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
2521 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
2581 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
2641 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
2701 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
2761 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
2821 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
2881 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
2941 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
3001 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
3061 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
3121 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
3181 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
3241 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
3301 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
3361 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
3421 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
3481 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
3541 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
3601 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
3661 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
3721 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
3781 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
3841 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
3901 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
3961 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
4021 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
4081 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
4141 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
4201 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
4261 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
4321 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
4381 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
4441 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
4501 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
4561 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
4621 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
4681 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
4741 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
4801 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
4861 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
4921 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
4981 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
5041 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
5101 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
5161 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
5221 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
5281 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
5341 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
5401 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
5461 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
5521 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
5581 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
5641 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
5701 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
5761 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
5821 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
5881 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
5941 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
6001 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
6061 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
6121 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
6181 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
6241 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
6301 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
6361 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
6421 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
6481 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
6541 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
6601 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
6661 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
6721 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
6781 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg c
//
