| 质粒名称: | pLacO-ISce1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 58505 |
| 存储实验室: | Tom Misteli |
|
载体骨架基本信息 |
|
| 载体名称: | pBluescript II KS |
| 空载体大小: | 3500 bp |
| 完整质粒大小: | 13500 bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 30 ℃ |
| 宿主菌: | stbl2 |
| 高拷贝/低拷贝: | 不清楚 |
| 培养基指南: | 为了避免质粒发生重组，请务必使用30度培养。该质粒，比较容易发生重组事件。 |
|
插入基因信息 |
|
| 插入基因名称: | Lac repeats adjacent to 6 copies of the ISce1 restriction site |
| 物种来源: | bacterial lac operator repeats |
| 插入大小: | 10000 bp |
| 启动子: | 无 |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | XhoI(未破坏) |
| 3’克隆位点: | XhoI(未破坏) |
| 5’测序引物 | M13 pUC forward primer |
| 3’测序引物 | M13pUC reverse primer |
|
文献引用方法 |
|
| 材料和方法部分: | pLacO-ISce1 was a gift from Tom Misteli (Addgene plasmid # 58505 ; http://n2t.net/addgene:58505 ; RRID:Addgene_58505) |
| 参考文献部分: |
Spatial dynamics of chromosome translocations in living cells. Roukos V, Voss TC, Schmidt CK, Lee S, Wangsa D, Misteli T. Science. 2013 Aug 9;341(6146):660-4. doi: 10.1126/science.1237150. 10.1126/science.1237150 PubMed 23929981 |
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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 pLacO-ISce1 3401 bp ds-DNA circular SYN 08-SEP-2021
DEFINITION provides 256 Lac operator repeats adjacent to the ISce1 restriction
site.
ACCESSION .
VERSION .
KEYWORDS pLacO-ISce1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3401)
AUTHORS Roukos V, Voss TC, Schmidt CK, Lee S, Wangsa D, Misteli T
TITLE Spatial dynamics of chromosome translocations in living cells.
JOURNAL Science. 2013 Aug 9;341(6146):660-4. doi: 10.1126/science.1237150.
PUBMED 23929981
REFERENCE 2 (bases 1 to 3401)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 8, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..3401
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 2..18
/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)."
protein_bind 38..54
/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)."
protein_bind 74..90
/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)."
protein_bind 110..126
/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)."
protein_bind 146..162
/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)."
protein_bind 182..198
/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)."
protein_bind 218..234
/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 complement(462..478)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(462..478)
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(515..535)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(515..533)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(554..570)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(554..570)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(567..589)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 578..594
/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(602..632)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 647..668
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(785..802)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(956..1544)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1036..1055)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1715..2575)
/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 2338..2357
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2576..2680)
/gene="bla"
/label=AmpR promoter
rep_origin 2707..3163
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2794..2813)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3004..3025
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3289..3311
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 3303..3320
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 3304..3320
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3330..3349
/label=T7
/note="T7 promoter, forward primer"
promoter 3330..3348
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 attgtgagcg gataacaatt tgtggccaca tgtggaattg tgagcggata acaatttgtg
61 gccacatgtg gaattgtgag cggataacaa tttgtggcca catgtggaat tgtgagcgga
121 taacaatttg tggccacatg tggaattgtg agcggataac aatttgtggc cacatgtgga
181 attgtgagcg gataacaatt tgtggccaca tgtggaattg tgagcggata acaatttgtg
241 gaattcggcg cctctagagt cgatccctcg aggtcgatag ggataacagg gtaatgtcga
301 gtagggataa cagggtaatg tcgagtaggg ataacagggt aatgtcgagt agggataaca
361 gggtaatgtc gagtagggat aacagggtaa tgtcgagtag ggataacagg gtaatgtcga
421 cggtatcgat aagcttgata tcgaattcct gcagcccggg ggatccacta gttctagagc
481 ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct
541 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
601 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
661 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
721 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
781 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
841 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
901 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
961 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
1021 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
1081 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
1141 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
1201 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
1261 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
1321 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
1381 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
1441 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
1501 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
1561 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
1621 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
1681 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
1741 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
1801 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
1861 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
1921 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
1981 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
2041 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
2101 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
2161 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
2221 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
2281 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
2341 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
2401 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
2461 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
2521 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
2581 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
2641 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
2701 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
2761 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
2821 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
2881 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
2941 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
3001 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
3061 atttataagg gattttgccg atttcggcct attggttaaa aaaatgagct gatttaacaa
3121 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttccattcg ccattcaggc
3181 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
3241 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
3301 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta
3361 ccgggccccc cctcgaggga tcgaggaatt ccacatgtgg a
//
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