| 质粒名称: | hSTARR-seq_ORI vector |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 99296 |
| 存储实验室: | Alexander Stark |
|
载体骨架基本信息 |
|
| 载体名称: | hSTARR-seq_ORI vector |
| 载体出品公司: | Promega |
| 空载体大小: | 2537 bp |
| 完整质粒大小: | 4010bp |
| 载体类型: | 哺乳细胞表达、hSTARR-seq screening vector |
| 载体抗性: | Chloramphenicol and Ampicillin |
| 生长温度: | 37 ℃ |
| 宿主菌: | ccdB Survival |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | 无 |
|
文献引用方法 |
|
| 材料和方法部分: | hSTARR-seq_ORI vector was a gift from Alexander Stark (Addgene plasmid # 99296 ; http://n2t.net/addgene:99296 ; RRID:Addgene_99296) |
| 参考文献部分: | Resolving systematic errors in widely used enhancer activity assays in human cells. Muerdter F, Boryn LM, Woodfin AR, Neumayr C, Rath M, Zabidi MA, Pagani M, Haberle V, Kazmar T, Catarino RR, Schernhuber K, Arnold CD, Stark A. Nat Methods. 2017 Dec 11. pii: nmeth.4534. doi: 10.1038/nmeth.4534. 10.1038/nmeth.4534 PubMed 29256496 |
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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 hSTARR-seq_ORI vector 4010 bp ds-DNA circular SYN 04-OCT-2023
DEFINITION Vector to measure enhancer activity of candidates from a DNA library
by STARR-seq..
ACCESSION .
VERSION .
KEYWORDS hSTARR-seq_ORI vector
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4010)
AUTHORS Muerdter F, Boryn LM, Woodfin AR, Neumayr C, Rath M, Zabidi MA,
Pagani M, Haberle V, Kazmar T, Catarino RR, Schernhuber K, Arnold
CD, Stark A
TITLE Resolving systematic errors in widely used enhancer activity assays
in human cells.
JOURNAL Nat Methods. 2017 Dec 11. pii: nmeth.4534. doi: 10.1038/nmeth.4534.
PUBMED 29256496
REFERENCE 2 (bases 1 to 4010)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Oct 4, 2023 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4010
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 1..861
/codon_start=1
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin complement(1051..1639)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1131..1150)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
intron 1735..1867
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
primer_bind 1850..1869
/label=pMT2-F
/note="Synthetic intron, forward primer"
promoter 2002..2032
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 2086..2745
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(2152..2171)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS 3087..3392
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind 3271..3290
/label=ccdB-fwd
/note="ccdB gene, forward primer"
polyA_signal 3470..3591
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3481..3500)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 3535..3554
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(3836..3853)
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 atgagcatcc aacattttcg tgtcgcactc attcccttct ttgcggcatt ttgcttgcct
61 gtttttgcac accccgaaac gctggtgaaa gtaaaagatg ctgaagatca actgggtgca
121 agagtgggct atatcgaact ggatctcaat agcggcaaga tccttgagtc tttccgcccc
181 gaagaacgat tcccgatgat gagcactttc aaagtactgc tatgtggcgc ggtgttgtcc
241 cgtatagacg ccgggcaaga gcagcttggt cgccgtatac actactcaca aaacgacttg
301 gttgagtact cgccggtcac ggaaaagcat cttacggatg gcatgacggt aagagaattg
361 tgtagtgctg ccattaccat gagcgacaac accgcggcca acttacttct gacaacgatc
421 ggaggcccta aggagctgac tgcatttctt cataatatgg gtgatcatgt gacccggctt
481 gaccgctggg aaccagagtt gaacgaagcc ataccgaacg acgagcgtga taccacgatg
541 ccagtagcaa tggccacaac tcttcggaaa ctactcactg gcgaacttct tactctagca
601 tcacgacagc agctcataga ctggatggag gcggacaaag tagcaggacc acttcttcgc
661 tcggccctcc ctgctggctg gttcattgct gacaaatcgg gggccggtga acgcggctct
721 cgcggcatca ttgctgcgct ggggcctgat ggtaagccct cacgaatcgt agtgatctac
781 acgacgggga gtcaggccac tatggacgaa cgaaataggc agatcgctga gatcggtgcc
841 tcactgatca agcactggta accactgcag tggtttagca tttgcggccg ctgtcagacc
901 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
961 aggtgaagat cctttttgat aatctcatga catgtgagca aaaggccagc aaaaggccag
1021 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
1081 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
1141 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
1201 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
1261 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
1321 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
1381 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
1441 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
1501 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
1561 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
1621 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
1681 gaacgaaaac tcacgttaag ggattttggg gcccggtcgt gaggcactgg gcaggtaagt
1741 atcaaggtta caagacaggt ttaaggagac caatagaaac tgggcttgtc gagacagaga
1801 agactcttgc gtttctgata ggcacctatt ggtcttactg acatccactt tgcctttctc
1861 tccacaggtg tccaccctgc aggcttaagc atggctagca aaggagaaga actcttcact
1921 ggagttgtcc caattcttgt tgaattagat tgatctagag catgcaccgg tgatatcgcg
1981 gccgcattag gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt
2041 ttgagttagg atccgtcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
2101 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
2161 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
2221 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
2281 ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg
2341 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
2401 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
2461 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
2521 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
2581 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
2641 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
2701 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaaacgcg tggatccggc
2761 ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata
2821 tatactgata tgtatacccg aagtatgtca aaaagaggta tgctatgaag cagcgtatta
2881 cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc
2941 cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa
3001 agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt
3061 tgctgacgag aacaggggct ggtgaaatgc agtttaaggt ttacacctat aaaagagaga
3121 gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga
3181 tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc
3241 cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc
3301 cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa
3361 acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt
3421 ctgcaggata tcgtcgacga attcggccgg ccgcttcgag cagacatgat aagatacatt
3481 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
3541 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
3601 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
3661 taaaacctct acaaatgtgg taaaatcgat aaggatccga ccgatgccct tgagagcctt
3721 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
3781 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc
3841 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
3901 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tcttgagaga
3961 cgtactagta accctgataa atgcttcaat aatattgaaa aaggacgagt
//
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