| 质粒名称: | hHS1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 159169 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3-eGFP |
| 载体出品公司: | Doug Golenbock (Addgene #13031) |
| 空载体大小: | 6146 bp |
| 完整质粒大小: | 7906 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | BleoR |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | hHS1 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1760 bp |
| 启动子: | CMV |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BamHI(未破坏) |
| 3’克隆位点: | XhoI (未破坏) |
| 5’测序引物 | CMV-F or T7 |
|
文献引用方法 |
|
| 材料和方法部分: |
hHS1 was a gift from Amit Reddi (Addgene plasmid # 159169 ; http://n2t.net/addgene:159169 ; RRID:Addgene_159169)
|
| 参考文献部分: | Heme dynamics and trafficking factors revealed by genetically encoded fluorescent heme sensors. Hanna DA, Harvey RM, Martinez-Guzman O, Yuan X, Chandrasekharan B, Raju G, Outten FW, Hamza I, Reddi AR. Proc Natl Acad Sci U S A. 2016 Jul 5;113(27):7539-44. doi: 10.1073/pnas.1523802113. Epub 2016 May 31. 10.1073/pnas.1523802113 PubMed 27247412 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS hHS1 6731 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Human codon-optimized cytosolic labile heme reporter.
ACCESSION .
VERSION .
KEYWORDS hHS1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6731)
AUTHORS Hanna DA, Harvey RM, Martinez-Guzman O, Yuan X, Chandrasekharan B,
Raju G, Outten FW, Hamza I, Reddi AR
TITLE Heme dynamics and trafficking factors revealed by genetically
encoded fluorescent heme sensors.
JOURNAL Proc Natl Acad Sci U S A. 2016 Jul 5;113(27):7539-44. doi:
10.1073/pnas.1523802113. Epub 2016 May 31.
PUBMED 27247412
REFERENCE 2 (bases 1 to 6731)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6731
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 141..520
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 521..724
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 675..695
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 769..788
/label=T7
/note="T7 promoter, forward primer"
promoter 769..787
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2645..2662)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2651..2875
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2921..3349
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3008..3027)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3218..3239
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3358..3378)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3363..3692
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3543..3678
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3605..3624
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 3740..3787
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 3806..4180
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 4310..4431
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4347..4366)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4401..4420
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4480..4496)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4480..4496)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4493..4515)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4504..4520
/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(4528..4558)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4573..4594
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4711..4728)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4882..5470)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4962..4981)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5641..6501)
/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 6264..6283
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6502..6606)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6681..6700)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
61 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
121 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
181 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
241 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
301 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
361 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
421 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
481 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
541 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
601 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
661 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
721 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca
781 ctatagggag acccaagctg gctagcgttt aaacttaagc ttggtaccga gctcggatcc
841 gccaccatgc acatggtcag cgagctgatc aaggaaaaca tgcacatgaa actgtacatg
901 gaggggactg tgaacaatca ccatttcaaa tgcacctccg agggcgaagg gaagccctac
961 gagggcacac agactatgag gatcaaggca gtggagggag gaccactgcc attcgccttt
1021 gacattctgg ctacctcatt catgtacggc agcaaaacct tcatcaatca cactcagggg
1081 attcccgact tctttaagca gtctttccct gaaggcttta cttgggagcg agtgaccaca
1141 tacgaggatg gaggcgtcct gaccgccaca caggacacaa gtctgcagga tggctgtctg
1201 atctataacg tgaagattcg cggggtcaac tttcccagta atggacctgt gatgcagaag
1261 aaaaccctgg gatgggaggc ttcaactgaa accctgtacc cagcagacgg aggactggag
1321 ggacgagcag atatggctct gaaactggtg ggcgggggac acctgatctg caacctgaag
1381 actacctatc ggtccaagaa acctgctaag aatctgaaaa tgccaggcgt gtactatgtg
1441 gaccggagac tggagagaat taaggaagca gataaagaga cctacgtgga gcagcacgaa
1501 gtggctgtcg cacgatattg tgacctgcct tctaaactgg gccatcgggg cgggtctatg
1561 gtgagtaagg gcgaggaact gttcacaggg gtggtcccaa tcctggtgga actggacggc
1621 gatgtcaatg ggcacaagtt cagcgtgtcc ggagagggag aaggggacgc aacctttgga
1681 ggcagcgccg acctggaaga taatatggag acactgaacg ataatctgaa agtgatcgag
1741 aaagccgaca acgccgctca ggtcaaggat gctctgacta aaatgagggc agccgctctg
1801 gatgcacaga aagccacccc ccctaagctg gaagacaaat cacctgatag cccagagatg
1861 aaggacttcc gccacggatt tgatatcctg gtcggccaga ttgacgatgc tctgaagctg
1921 gcaaatgaag gcaaggtgaa agaggcacag gcagccgctg agcagctgaa aacaactagg
1981 aacgcctacc atcagaagta tcgcggggga aagctgacac tgaaattcat ctgcaccaca
2041 ggcaagctgc ccgtgccctg gccaactctg gtcactaccc tgggatacgg cgtgcagtgt
2101 ttttcccgct atccagacca catgaagcag catgatttct ttaaatctgc catgcccgaa
2161 ggctacgtgc aggagagaac catcttcttt aaggacgatg gaaactataa aacaagggct
2221 gaagtgaagt tcgagggaga cactctggtc aaccgcatcg aactgaaggg cattgacttt
2281 aaagaggatg gaaatattct gggccacaag ctggaataca actataatag ccataacgtg
2341 tacatcatgg ccgataagca gaaaaacggc attaaggtca atttcaaaat ccggcacaat
2401 attgaggacg ggagcgtgca gctggccgat cattaccagc agaacacccc aatcggggac
2461 ggaccagtgc tgctgcccga taatcactat ctgtccacac agtctgccct gagtaaggac
2521 cctaacgaaa aaagagatca catggtgctg ctggagtttg tcaccgcagc cgggattaca
2581 ctgggaatgg acgagctgta caagtgactc gagtctagag ggcccgttta aacccgctga
2641 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
2701 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
2761 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
2821 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
2881 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
2941 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
3001 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
3061 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
3121 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
3181 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
3241 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
3301 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
3361 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
3421 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
3481 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
3541 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
3601 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
3661 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
3721 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
3781 atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac
3841 cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga
3901 cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc
3961 ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga
4021 cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc
4081 ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc
4141 cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga
4201 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
4261 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
4321 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
4381 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
4441 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
4501 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
4561 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
4621 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
4681 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
4741 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
4801 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
4861 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
4921 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
4981 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
5041 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
5101 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
5161 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
5221 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
5281 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
5341 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
5401 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
5461 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
5521 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
5581 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
5641 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
5701 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
5761 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
5821 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
5881 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
5941 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
6001 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
6061 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
6121 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
6181 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
6241 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
6301 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
6361 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
6421 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
6481 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
6541 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
6601 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
6661 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
6721 ctccctgctt g
//
