LOCUS pCS2+HyPer7-NLS 5615 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Mammalian expression of nucleus targeted ultrasensitive hydrogen
peroxide indicator HyPer7 for optical imaging.
ACCESSION .
VERSION .
KEYWORDS pCS2+HyPer7-NLS
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5615)
AUTHORS Pak VV, Ezerina D, Lyublinskaya OG, Pedre B, Tyurin-Kuzmin PA,
Mishina NM, Thauvin M, Young D, Wahni K, Martinez Gache SA,
Demidovich AD, Ermakova YG, Maslova YD, Shokhina AG, Eroglu E, Bilan
DS, Bogeski I, Michel T, Vriz S, Messens J, Belousov VV
TITLE Ultrasensitive Genetically Encoded Indicator for Hydrogen Peroxide
Identifies Roles for the Oxidant in Cell Migration and Mitochondrial
Function.
JOURNAL Cell Metab. 2020 Mar 3;31(3):642-653.e6. doi:
10.1016/j.cmet.2020.02.003.
PUBMED 32130885
REFERENCE 2 (bases 1 to 5615)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
//www.snapgene.com
FEATURES Location/Qualifiers
source 1..5615
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 35..53
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 35..52
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind 717..738
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(847..868)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1108..1127)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
CDS 1552..1572
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1576..1596
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1600..1620
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
promoter complement(1645..1662)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 1667..1801
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1691..1710)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 1745..1764
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(1914..1934)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(1914..1932)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(1953..1969)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1953..1969)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1966..1988)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1977..1993
/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(2001..2031)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2046..2067
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2184..2201)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2355..2943)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2435..2454)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3114..3974)
/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 3737..3756
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3975..4079)
/gene="bla"
/label=AmpR promoter
rep_origin complement(4105..4560)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4242..4263)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4454..4473
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4638..5615
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
ORIGIN
1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
61 ctacttgttc tttttgcagg atcccatcga ttcgaattcg ccaccatgca cctggctaat
121 gaggagcaaa acgaattgga aggtgctttt aagctgggct tgatctttac cgtcgccccg
181 tacttgctgc ctaagttgat cgtcagcctg cgccgcaccg ccccgaagat gcctttgatg
241 ttggaggaga attacaccca cacccttacc gagtcactga aacgcggcga cgttgacgcc
301 atcattgtgg cagagccgtt tcaagagccg ggtattgtga ctgaaccttt gtatgacgag
361 ccgtttttcg ttatcgttcc gaagggccat agcttcgagg agttggatgc cgttagccct
421 cgcatgttgg gcgaagaaca agttctgctt ctgaccgagg gcaactgtat gcgtgaccag
481 gtgcttagct ccgccggctt caacagcaac aacgtctata tcatggccga caagcagaag
541 aacggcatca aggccaactt caagatccgc cacaacgtcg aggacggcag cgtgcagctc
601 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
661 cactacctga gcttccagtc cgtcctgagc aaagacccca acgagaagcg cgatcacatg
721 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaac
781 gtggatggcg gtagcggtgg caccggcagc aagggcgagg agctgttcac cggggtggtg
841 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
901 ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag
961 ctgcccgtgc cctggcccac cctcgtgacc accctcagct acggcctgaa gtgcttcgcc
1021 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
1081 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
1141 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcgg cttcaaggag
1201 gacggcaaca tcctggggca caagctggag tacaacggcc ccagttgcag cgagcttgcc
1261 gccaaacagc gcattcaggg tctcaccaac acccttcaag gctcaagcat caataccatc
1321 cgccacatgg ttgccagtgg cttggcaatc agcgtcttgc ctgccactgc ccttaccgaa
1381 aacgaccaca tgctgtttag cattatcccg tttgaaggca ccccgcctag ccgccgcgtt
1441 gtcctggcct atcgccgcaa ctttgtccgc cctaaggccc tttcagccat gaaagccgca
1501 attatgcaaa gccaactgca tggtgttagc ttcatctgcg ataccggtga tccaaaaaag
1561 aagagaaagg tagatccaaa aaagaagaga aaggtagatc caaaaaagaa gagaaaggta
1621 taagcttgcc tcgagcctct agaactatag tgagtcgtat tacgtagatc cagacatgat
1681 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
1741 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
1801 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
1861 ttaattcgcg gccgcggcgc caatgcattg ggcccggtac ccagcttttg ttccctttag
1921 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
1981 tatccgctca caattccaca caacatacga gccgggagca taaagtgtaa agcctggggt
2041 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
2101 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
2161 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
2221 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
2281 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
2341 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
2401 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
2461 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
2521 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
2581 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
2641 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
2701 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
2761 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
2821 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
2881 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
2941 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
3001 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
3061 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
3121 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
3181 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
3241 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
3301 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
3361 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
3421 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
3481 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
3541 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
3601 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
3661 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
3721 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
3781 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
3841 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
3901 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
3961 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
4021 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
4081 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
4141 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
4201 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
4261 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
4321 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
4381 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
4441 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
4501 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
4561 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
4621 tattacgcca gtcgaccgcc aattcaatat ggcgtatatg gactcatgcc aattcaatat
4681 ggtggatctg gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc aattcaatat
4741 ggtggatctg gaccccagcc aattcaatat ggcggacttg gcaccatgcc aattcaatat
4801 ggcggacctg gcactgtgcc aactggggag gggtctactt ggcacggtgc caagtttgag
4861 gaggggtctt ggccctgtgc caagtccgcc atattgaatt ggcatggtgc caataatggc
4921 ggccatattg gctatatgcc aggatcaata tataggcaat atccaatatg gccctatgcc
4981 aatatggcta ttggccaggt tcaatactat gtattggccc tatgccatat agtattccat
5041 atatgggttt tcctattgac gtagatagcc cctcccaatg ggcggtccca tataccatat
5101 atggggcttc ctaataccgc ccatagccac tcccccattg acgtcaatgg tctctatata
5161 tggtctttcc tattgacgtc atatgggcgg tcctattgac gtatatggcg cctcccccat
5221 tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc aataggacca
5281 cccaccattg acgtcaatgg gatggctcat tgcccattca tatccgttct cacgccccct
5341 attgacgtca atgacggtaa atggcccact tggcagtaca tcaatatcta ttaatagtaa
5401 cttggcaagt acattactat tggaagtacg ccagggtaca ttggcagtac tcccattgac
5461 gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca tccccattga cgtcaatggg
5521 gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg cgtgcctaat
5581 gggaggtcta tataagcaat gctcgtttag ggaac
//
