LOCUS pCS2+IMS-HyPer7 5765 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Mammalian expression of mitochondrial intermembrane space targeted
ultrasensitive hydrogen peroxide indicator HyPer7 for optical
imaging.
ACCESSION .
VERSION .
KEYWORDS pCS2+IMS-HyPer7
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5765)
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 5765)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
//www.snapgene.com
FEATURES Location/Qualifiers
source 1..5765
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 475..493
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 475..492
/label=SP6
/note="SP6 promoter, forward primer"
regulatory 540..549
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
regulatory 544..553
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind 1391..1412
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(1521..1542)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1782..1801)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
promoter complement(2235..2252)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 2257..2391
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2281..2300)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2335..2354
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(2504..2524)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(2504..2522)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(2543..2559)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2543..2559)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2556..2578)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2567..2583
/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(2591..2621)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2636..2657
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2774..2791)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2945..3533)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3025..3044)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3704..4564)
/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 4327..4346
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4565..4669)
/gene="bla"
/label=AmpR promoter
rep_origin complement(4695..5150)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4832..4853)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5044..5063
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 5228..440
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
ORIGIN
1 acgtcatatg ggcggtccta ttgacgtata tggcgcctcc cccattgacg tcaattacgg
61 taaatggccc gcctggctca atgcccattg acgtcaatag gaccacccac cattgacgtc
121 aatgggatgg ctcattgccc attcatatcc gttctcacgc cccctattga cgtcaatgac
181 ggtaaatggc ccacttggca gtacatcaat atctattaat agtaacttgg caagtacatt
241 actattggaa gtacgccagg gtacattggc agtactccca ttgacgtcaa tggcggtaaa
301 tggcccgcga tggctgccaa gtacatcccc attgacgtca atggggaggg gcaatgacgc
361 aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc ctaatgggag gtctatataa
421 gcaatgctcg tttagggaac cgccattctg cctggggacg tcggagcaag cttgatttag
481 gtgacactat agaatacaag ctacttgttc tttttgcagg atcccatcga ttcgaattcg
541 ccaccatggc ggctctgaag agttggctgt cgcgcagcgt aacttcattc ttcaggtaca
601 gacagtgttt gtgtgttcct gttgtggcta actttaagaa gcggtgtttc tcagaattga
661 taagaccatg gcacaaaact gtgacgattg gctttggagt aaccctgtgt gcggttccta
721 ttaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccaccc gtcgccaccg
781 gtcacctggc taatgaggag caaaacgaat tggaaggtgc ttttaagctg ggcttgatct
841 ttaccgtcgc cccgtacttg ctgcctaagt tgatcgtcag cctgcgccgc accgccccga
901 agatgccttt gatgttggag gagaattaca cccacaccct taccgagtca ctgaaacgcg
961 gcgacgttga cgccatcatt gtggcagagc cgtttcaaga gccgggtatt gtgactgaac
1021 ctttgtatga cgagccgttt ttcgttatcg ttccgaaggg ccatagcttc gaggagttgg
1081 atgccgttag ccctcgcatg ttgggcgaag aacaagttct gcttctgacc gagggcaact
1141 gtatgcgtga ccaggtgctt agctccgccg gcttcaacag caacaacgtc tatatcatgg
1201 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac gtcgaggacg
1261 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
1321 tgctgcccga caaccactac ctgagcttcc agtccgtcct gagcaaagac cccaacgaga
1381 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
1441 acgagctgta caacgtggat ggcggtagcg gtggcaccgg cagcaagggc gaggagctgt
1501 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
1561 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct
1621 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctc agctacggcc
1681 tgaagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
1741 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
1801 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
1861 tcggcttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac ggccccagtt
1921 gcagcgagct tgccgccaaa cagcgcattc agggtctcac caacaccctt caaggctcaa
1981 gcatcaatac catccgccac atggttgcca gtggcttggc aatcagcgtc ttgcctgcca
2041 ctgcccttac cgaaaacgac cacatgctgt ttagcattat cccgtttgaa ggcaccccgc
2101 ctagccgccg cgttgtcctg gcctatcgcc gcaactttgt ccgccctaag gccctttcag
2161 ccatgaaagc cgcaattatg caaagccaac tgcatggtgt tagcttcatc tgcgattgac
2221 tcgagcctct agaactatag tgagtcgtat tacgtagatc cagacatgat aagatacatt
2281 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
2341 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
2401 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaattcgcg
2461 gccgcggcgc caatgcattg ggcccggtac ccagcttttg ttccctttag tgagggttaa
2521 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
2581 caattccaca caacatacga gccgggagca taaagtgtaa agcctggggt gcctaatgag
2641 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
2701 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
2761 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
2821 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
2881 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
2941 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
3001 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
3061 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
3121 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
3181 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
3241 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
3301 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
3361 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
3421 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
3481 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
3541 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
3601 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
3661 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
3721 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
3781 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
3841 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
3901 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
3961 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
4021 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
4081 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
4141 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
4201 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
4261 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
4321 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
4381 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
4441 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
4501 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
4561 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
4621 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
4681 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
4741 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
4801 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
4861 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
4921 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
4981 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
5041 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
5101 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
5161 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
5221 gtcgaccgcc aattcaatat ggcgtatatg gactcatgcc aattcaatat ggtggatctg
5281 gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc aattcaatat ggtggatctg
5341 gaccccagcc aattcaatat ggcggacttg gcaccatgcc aattcaatat ggcggacctg
5401 gcactgtgcc aactggggag gggtctactt ggcacggtgc caagtttgag gaggggtctt
5461 ggccctgtgc caagtccgcc atattgaatt ggcatggtgc caataatggc ggccatattg
5521 gctatatgcc aggatcaata tataggcaat atccaatatg gccctatgcc aatatggcta
5581 ttggccaggt tcaatactat gtattggccc tatgccatat agtattccat atatgggttt
5641 tcctattgac gtagatagcc cctcccaatg ggcggtccca tataccatat atggggcttc
5701 ctaataccgc ccatagccac tcccccattg acgtcaatgg tctctatata tggtctttcc
5761 tattg
//
