| 质粒名称: | pDonorCm(Ptr) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 175577 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pCDFDuet-1 |
| 完整质粒大小: | 4086 bp |
| 载体类型: | 细菌细胞表达 |
| 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: | Mini-Tn, Cm cassette |
| 物种来源: | Pseudoalteromonas translucida |
| 基因突变: | NO |
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: |
pDonorCm(Ptr) was a gift from Sheng Yang (Addgene plasmid # 175577 ; http://n2t.net/addgene:175577 ; RRID:Addgene_175577)
|
| 参考文献部分: | Orthogonal CRISPR-associated transposases for parallel and multiplexed chromosomal integration. Yang S, Zhang Y, Xu J, Zhang J, Zhang J, Yang J, Jiang Y, Yang S. Nucleic Acids Res. 2021 Sep 3. pii: 6363765. doi: 10.1093/nar/gkab752. 10.1093/nar/gkab752 PubMed 34478496 |
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
质粒图谱.png)
载体简介
载体序列
LOCUS pDonorCm(Ptr) 4086 bp ds-DNA circular SYN 03-NOV-2021
DEFINITION DNA donor for transposition.
ACCESSION .
VERSION .
KEYWORDS pDonorCm(Ptr)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4086)
AUTHORS Yang S, Zhang Y, Xu J, Zhang J, Zhang J, Yang J, Jiang Y, Yang S
TITLE Orthogonal CRISPR-associated transposases for parallel and
multiplexed chromosomal integration.
JOURNAL Nucleic Acids Res. 2021 Sep 3. pii: 6363765. doi:
10.1093/nar/gkab752.
PUBMED 34478496
REFERENCE 2 (bases 1 to 4086)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 3, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4086
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 198..217
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 451..468
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 585..606
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 621..651
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 659..675
/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 664..686
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 683..699
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 683..699
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 831..1490
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(897..916)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
primer_bind complement(2169..2186)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(2169..2185)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2178..2200)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(2394..2413)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2513..2535
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(2573..2591)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 2659..2763
/gene="bla"
/label=AmpR promoter
CDS 2764..3624
/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 complement(2982..3001)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3795..297
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
61 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
121 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
181 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
241 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
301 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
361 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
421 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
481 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
541 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
601 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
661 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttttg
721 ggtgttgttt gaagtataag ttgacatatc tgtactaaaa gatggcataa attggaagtg
781 taaggtggca tagtctagta tttaaccaaa tggttaaatg gttgactcac atggagaaaa
841 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg
901 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct
961 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg
1021 cccgcctgat gaatgctcat ccggagttcc gtatggcaat gaaagacggt gagctggtga
1081 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat
1141 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg
1201 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt
1261 tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg
1321 acaacttctt cgcccccgtt ttcactatgg gcaaatatta tacgcaaggc gacaaggtgc
1381 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
1441 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag
1501 gcagttattg gtgccctttt aattttcctt aattattttt aaagttagac tgatttagac
1561 ttggaaaagc ttaatgattg gagagctaaa ttgactaata gtattcagtc aagtttaatt
1621 agttttaagc gatccatgca taactatttc tgtacaatgc tatattttca ccaattaata
1681 atttttaatc aagcctacat tatgaaatat actataccca tttgaactct tctatttgta
1741 ccattgtcgg tagcaaaaac ttatggggtt ttgaacgtca cttaaattgt aagcatttgc
1801 gatggaggcg cgtttagagt caaccttgat tctgatatgc tccgaatttt tggtaagaat
1861 ataagtgtga gagtagctaa tgtggatacg cctgagttaa gggaaaaatg tgaaaatgaa
1921 ataactcgtt atcatgcaaa gtgactaagg ttataatctt ccgtttatgg cacatagcag
1981 ccaactaaac ttgacagtat ttttatgtgg ttggctttat aaaaccagca tttggtaaca
2041 ttatgccaat ttttacttca atattatgcc aacatacact acactaacgg agctgtagca
2101 caataagctc gtttgtactt atgccaactt atacttcaaa caacattggg ggtaccgagc
2161 tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
2221 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
2281 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
2341 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
2401 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
2461 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
2521 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
2581 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
2641 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
2701 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
2761 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
2821 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
2881 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
2941 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
3001 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
3061 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
3121 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
3181 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
3241 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
3301 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
3361 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
3421 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
3481 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
3541 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
3601 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
3661 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
3721 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
3781 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
3841 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
3901 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
3961 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
4021 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
4081 tagtta
//
