LOCUS pcDNA3.1-Peredox-mCherry 8128 bp DNA circular SYN 25-AUG-2024
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8128)
AUTHORS Hung YP, Albeck JG, Tantama M, Yellen G
TITLE Imaging Cytosolic NADH-NAD(+) Redox State with a Genetically Encoded
Fluorescent Biosensor.
JOURNAL Cell Metab. 2011 Oct 5;14(4):545-54.
PUBMED 21982714
REFERENCE 2 (bases 1 to 8128)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell
Metab."; date: "2011-10-5"; volume: "14(4)"; pages: "545-54"
FEATURES Location/Qualifiers
source 1..8128
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(9..29)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 184..563
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind 735..754
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 829..933
/gene="bla"
/label=AmpR promoter
CDS 934..1794
/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 complement(1152..1171)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1965..2553
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2454..2473
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2707..2724
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 2841..2862
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2877..2907
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2915..2931
/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)."
primer_bind 2920..2942
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2939..2955
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2939..2955
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
polyA_signal 3004..3125
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3015..3034)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 3069..3088
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(3299..4093)
/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(3410..3429)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 4020..4039
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
promoter complement(4160..4489)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4174..4309
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(4228..4247)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 4474..4494
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin complement(4503..4931)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4613..4634)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4825..4844
/label=F1ori-R
/note="F1 origin, reverse primer"
polyA_signal 4977..5201
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 5190..5207
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
CDS complement(5240..5950)
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(5518..5537)
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 5787..5805
/label=mCherry-R
/note="mCherry, reverse primer"
CDS complement(5951..7996)
/codon_start=1
/product="genetically encoded fluorescent biosensor of the
cytosolic NADH-NAD+ redox state (Hung et al., 2011)"
/label=Peredox
/note="consists of two pH-insensitive mutant copies of the
Thermus aquaticus redox-sensing repressor Rex (T-Rex)
separated by circularly permuted monomeric T-Sapphire
(cpmTS), a pH-resistant GFP variant"
/translation="KVPEAAISRLITYLRILEELEAQGVHRTASEQLGELAQVTAFQVD
KDLSYFGSYGTDGVGYTVPVLKRELRHILGLNRKWGLCIVGMGRLGSALADWPGFGESF
ELRGFFDVDPGMVGRPVRGGVIEHVDLLPQRVPGRIEIALLTVPREAAQKAADLLVAAG
IKGILNFAPVVLEVPKEVAVENVDILAGLTRLSFAILNPTWSAAGGHGFTAHNVYIMAD
KQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSIQSKLSKDPNEK
RDHMVLLEFVTAAGITHGMDELYKGGTGGSMVSKGEELFTGVVPILVELDGDVNGHKFS
VSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFSYGVMVFARYPDHMKQHDFFKSA
MPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNTK
VPEAAISRLITYLRILEELEAQGVHRTASEQLGELAQVTAFQVDEDLSYFGSYGTDGVG
YTVPVLKRELRHILGLNRKWGLCIVGMGRLGSALADWPGFGESFELRGFFDVDPGMVGR
PVRGGVIEHVDLLPQRVPGRIEIALLTVPREAAQKAADLLVAAGIKGILNFAPVVLEVP
KEVAVENVDFLAGLTRLSFAILNPKWREEMMGSGTGGNASDGGGSGG"
primer_bind complement(8044..8063)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(8045..8063)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(join(8108..8128,1..183))
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
ORIGIN
1 ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg
61 aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact
121 tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat
181 caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa
241 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg
301 cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc
361 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat
421 tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta
481 tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg attactatta
541 ataactagtc aataatcaat gtcaacgcgt atatctggcc cgtacatcgc gaagcagcgc
601 aaaacgccta accctaagca gattcttcat gcaattgtcg gtcaagcctt gccttgttgt
661 agcttaaatt ttgctcgcgc actactcagc gacctccaac acacaagcag ggagcagata
721 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata ggggatcggg
781 agatctcccg atccgtcgac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
841 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
901 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
961 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
1021 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
1081 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
1141 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
1201 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
1261 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
1321 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
1381 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
1441 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
1501 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
1561 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
1621 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
1681 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
1741 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
1801 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
1861 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
1921 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
1981 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
2041 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
2101 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
2161 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
2221 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
2281 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
2341 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
2401 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
2461 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
2521 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
2581 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
2641 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
2701 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
2761 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
2821 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
2881 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
2941 ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacgg tatacagaca
3001 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
3061 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
3121 aagttggggt gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg
3181 gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa
3241 tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc
3301 agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac
3361 cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg
3421 tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc
3481 cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga
3541 cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg gctggcgcga
3601 gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac
3661 gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg
3721 tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag
3781 atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag
3841 tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg
3901 ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg
3961 ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg
4021 cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat
4081 cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagatccg aaaatggata
4141 tacaagctcc cgggagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact
4201 tctggaatag ctcagaggca gaggcggcct cggcctctgc ataaataaaa aaaattagtc
4261 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
4321 ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
4381 gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac
4441 ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagaa
4501 ttaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
4561 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
4621 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
4681 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
4741 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
4801 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
4861 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
4921 acagggcgcg tggggatacc ccctagagcc ccagctggtt ctttccgcct cagaagccat
4981 agagcccacc gcatccccag catgcctgct attgtcttcc caatcctccc ccttgctgtc
5041 ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc
5101 ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg
5161 ggaggggcaa acaacagatg gctggcaact agaaggcaca gtcgaggctg atcagcggtt
5221 taaacttaag cttctcgact cacttgtaca gttcgtccat gccgccggtg gagtggcggc
5281 cctcggcgcg ttcgtactgt tccacgatgg tgtagtcctc gttgtgggag gtgatgtcca
5341 acttgatgtt gacgttgtag gcgccgggca gctgcacggg cttcttggcc ttgtaggtgg
5401 tcttgacctc agcgtcgtag tggccgccgt ccttcagctt cagcctctgc ttgatctcgc
5461 ccttcagggc gccgtcctcg gggtacatcc gctcggagga ggcctcccag cccatggtct
5521 tcttctgcat tacggggccg tcggagggga agttggtgcc gcgcagcttc accttgtaga
5581 tgaactcgcc gtcctgcagg gaggagtcct gggtcacggt caccacgccg ccgtcctcga
5641 agttcatcac gcgctcccac ttgaagccct cggggaagga cagcttcaag tagtcgggga
5701 tgtcggcggg gtgcttcacg taggccttgg agccgtacat gaactgaggg gacaggatgt
5761 cccaggcgaa gggcaggggg ccacccttgg tcaccttcag cttggcggtc tgggtgccct
5821 cgtaggggcg gccctcgccc tcgccctcga tctcgaactc gtggccgttc acggagccct
5881 ccatgtgcac cttgaagcgc atgaactcct tgatgatggc catgttatcc tcctcgccct
5941 tgctcaccat accaccagac ccaccaccgt cagaggcgtt gcccccagtt ccggaaccca
6001 tcatttcttc gcgccactta ggattcagga ttgcgaagct caggcgggtc aggccggcca
6061 ggaaatcaac gttttcaaca gcaacttcct taggaacctc cagaacaaca ggggcgaagt
6121 tcagaattcc cttgatacca gcggcgacta gtaagtcggc tgccttctgt gcagcttcgc
6181 gaggaacggt cagcagggcg atctcgatgc gacccgggac gcgctgtggc aggaggtcaa
6241 catgctcgat aacgccaccg cgaacagggc gcccaaccat gcctggatca acgtcgaaga
6301 agccgcgcag ttcgaaagat tcaccaaagc ccggccagtc tgccagtgca ctaccgaggc
6361 ggcccatgcc aacgatgcat aagccccact tgcggttcag gcccaggatg tggcgcagct
6421 cgcgcttcag gacagggact gtgtagccaa caccgtcggt gccgtaggaa ccgaaatagg
6481 acagatcttc gtcaacctgg aatgctgtaa cctgtgccag ttcgcccagc tgctcgcttg
6541 cagtgcggtg aacgccttgg gcttcgagct cctccaggat gcggaggtaa gtgatgaggc
6601 ggctgatggc ggcttcaggt accttggtgt tatattcaag cttatggccc aggatgttgc
6661 cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt aaccagagtg tcgccctcga
6721 acttaacctc agcgcgtgtc ttatagttgc cgtcgtcctt gaaaaagatg gtgcgttcct
6781 gaacgtagcc ctcaggcatg gcgctcttga agaagtcgtg ctgcttcatg tggtcaggat
6841 atctggcgaa aaccataaca ccataggaga aggtagtaac cagggtaggc caaggaacag
6901 gcagcttgcc agtggtgcag atgaacttca gagtcagctt gccgtaggta gcgtctcctt
6961 cgccttcccc gctaacgctg aatttgtggc cgttaacgtc gccgtccagt tcaaccagga
7021 taggaacaac accagtgaac agttcctcgc ctttggaaac catagatccc ccggtgccgc
7081 ccttgtacag ttcgtccatg ccgtgggtga taccagcggc ggtaacaaat tccagcagaa
7141 ccatgtggtc gcgcttctcg tttgggtctt tggacagttt agactggatg cttaagtagt
7201 ggttgtcagg cagcagaaca gggccgtcgc cgattggggt gttctgctgg tagtgatcag
7261 ccagctgaac gccgccgtct tcgatgttgt ggcggatctt gaagttggcc ttgataccgt
7321 tcttctgttt gtcagccatg atatacacgt tgtgagcagt gaaaccatgc ccacctgctg
7381 cgctccacgt ggggttcaga atggcaaagc tcagacgcgt cagaccggcc aggatgtcaa
7441 cgttctcaac agccacttcc ttgggaacct ccagtacaac tggagcgaag ttcaggatgc
7501 ccttgatgcc agcggcaacc agcaggtcgg cagccttctg tgctgcctcg cgtggaaccg
7561 tcagcagagc gatttcgatg cgaccaggta cgcgctgggg cagcagatca acgtgttcga
7621 taacaccacc gcgaaccggc cgaccaacca tacctgggtc aacatcgaag aaaccgcgca
7681 gctcgaagct ctcgccaaaa ccaggccaat cagcaagagc ggatcccagg cggcccatcc
7741 ccacgatgca caggccccat ttgcggttca gaccgaggat atggcgcagt tctctcttga
7801 ggaccggtac agtgtagcca acgccgtcag tgccgtagga gccgaagtag gacaggtcct
7861 tatcaacctg gaaggcggtg acctgggcca gctcgcccag ttgctcggag gcggtgcggt
7921 gtacaccttg ggcttccagc tcttccagaa ttctcaggta agtaatcagt ctggaaatgg
7981 ctgcttcagg aactttgcta gcaaccatgg tgggcccaat ctagagctag ccagcttggg
8041 tctccctata gtgagtcgta ttaatttcga taagccagta agcagtgggt tctctagtta
8101 gccagagagc tctgcttata tagacctc
//
