| 质粒名称: | pBAV1K-T5-gfp |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 26702 |
| 存储实验室: | Ichiro Matsumura |
|
载体骨架基本信息 |
|
| 载体名称: | pBAV1K |
| 载体出品公司: | Clontech |
| 空载体大小: | 2792bp |
| 完整质粒大小: | 3653bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | Kanamycin |
| 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: | T5lacOlacOeGFP biobrick |
| 物种来源: | Synthetic construct |
| 插入大小: | 861bp |
| GenBank编号 | HQ191434 |
| 启动子: | CMV |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI, XbaI (not destroyed) |
| 3’克隆位点: | SpeI, PstI (not destroyed) |
| 5’测序引物 | GACGAACTCCAATTCACTGTTCCTTGC |
| 3’测序引物 | GGAGAGCGTTCACCGACAAACAACAG |
|
文献引用方法 |
|
| 材料和方法部分: |
pBAV1K-T5-gfp was a gift from Ichiro Matsumura (Addgene plasmid # 26702 ; http://n2t.net/addgene:26702 ; RRID:Addgene_26702)
|
| 参考文献部分: | Rational design of a plasmid origin that replicates efficiently in both gram-positive and gram-negative bacteria. Bryksin AV, Matsumura I. PLoS One. 2010 . 5(10):e13244. 10.1371/journal.pone.0013244 PubMed 20949038 |
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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 pBAV1K-T5-gfp 3653 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pBAV1K-T5-gfp
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3653)
AUTHORS Bryksin AV, Matsumura I
TITLE Rational design of a plasmid origin that replicates efficiently in
both gram-positive and gram-negative bacteria.
JOURNAL PLoS One. 2010 . 5(10):e13244.
PUBMED 20949038
REFERENCE 2 (bases 1 to 3653)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..3653
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 26..70
/label=T5 promoter
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 46..62
/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 78..94
/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)."
RBS 104..115
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
primer_bind complement(152..180)
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind 775..795
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
misc_feature 842..862
/label=BioBrick suffix
/note="universal suffix for all parts"
terminator 913..999
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS 2330..2341
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
terminator 2543..2637
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(2653..3447)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEEYRCVKDTEGMSPAKVYKLVGENEILYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEGYED
EQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 3653..21
/label=BioBrick prefix
/note="BioBrick prefix for parts that do not start with
""ATG"""
ORIGIN
1 aattcgcggc cgcttctaga ggaaatcata aaaaatttat ttgctttgtg agcggataac
61 aattataata gattcaattg tgagcggata acaattacta gagaaagagg agaaatacta
121 gatgcgtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg
181 tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg
241 aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact
301 tgtcactact ctgacgtatg gtgttcaatg cttttcccgt tatccggatc atatgaaacg
361 gtatgacttt ttcaagagtg ccatgcccga aggttatgta caggaacgca ctatatcttt
421 caaagatgac gggaactaca agacgcgtgc tgaagtcaag tttgaaggtg atacccttgt
481 taatcgtatc gagttaaaag gtattgattt taaagaagat ggaaacattc tcggacacaa
541 actcgagtac aactataact cacacaatgt atacatcacg gcagacaaac aaaagaatgg
601 aatcaaagct aacttcaaaa ttcgccacaa cattgaagat ggatccgttc aactagcaga
661 ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta
721 cctgtcgaca caatctgccc ttttgaaaga tcccaacgaa aagcgtgacc acatggtcct
781 tcttgagttt gtaactgctg ctgggattac acatggcatg gatgaactat acaaataata
841 atactagtag cggccgctgc aggcctcagg gcccgatcga tgccgccgct taattaatta
901 atccagaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
961 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgcccta gacctagtgt
1021 cattttattt cccccgtttc agcatcaaga acctttgcat aacttgctct atatccacac
1081 tgataattgc cctcaaacca taatctaaag gcgctagagt ttgttgaaac aatatctttt
1141 acatcattcg tatttaaaat tccaaactcc gctcccctaa ggcgaataaa agccattaaa
1201 tcttttgtat ttaccaaatt atagtcatcc actatatcta agagtaaatt cttcaattct
1261 cttttttggc tttcatcaag tgttatatag cggtcaatat caaaatcatt aatgttcaaa
1321 atatcttttt tgtcgtatat atgtttattc ttagcaatag cgtcctttga ttcatgagtc
1381 aaatattcat atgaaccttt gatataatca agtatctcaa catgagcaac tgaactattc
1441 cccaattttc gcttaatctt gttcctaacg ctttctattg ttacaggatt tcgtgcaata
1501 tatataacgt gatagtgtgg ttttttatag tgctttccat ttcgtataac atcactacta
1561 ttccatgtat ctttatcttt tttttcgtcc atatcgtgta aaggactgac agccatagat
1621 acgcccaaac tctctaattt ttccttccaa tcattaggaa ttgagtcagg atataataaa
1681 aatccaaaat ttctagcttt agtattttta atagccatga tataattacc ttatcaaaaa
1741 caagtagcga aaactcgtat ccttctaaaa acgcgagctt tcgcttattt tttttgttct
1801 gattcctttc ttgcatattc ttctatagct aacgccgcaa ccgcagattt tgaaaaacct
1861 ttttgtttcg ccatatctgt taatttttta tcttgctctt ttgtcagaga aatcataact
1921 ctttttttcg attctgaaat caccatttaa aaaactccaa tcaaataatt ttataaagtt
1981 agtgtatcac tttgtaatca taaaaacaac aataaagcta cttaaatata gatttataaa
2041 aaacgttggc gaaaacgttg gcgattcgtt ggcgattgaa aaacccctta aacccttgag
2101 ccagttggga tagagcgttt ttggcacaaa aattggcact cggcacttaa tggggggtcg
2161 tagtacggaa gcaaaattcg cttcctttcc ccccattttt ttccaaattc cagatttttt
2221 tcaaaaattt tccagcgcta ccgctcggca aaattgcaag caatttttaa aatcaaaccc
2281 atgagggaat ttcattccct catactccct tgagcctcct ccaaccgaaa tagaagggcg
2341 ctgcgcttat tatttcattc agtcatcggc tttcataatc taacagacaa catcttcgct
2401 gcaaagccac gctacgctca agggctttta cgctacgata acgcctgttt taacgattat
2461 gccgataact aaacgaaata aacgctaaaa cgtctcagaa acgattttga gacgttttaa
2521 taaaaaatcg cctagtgctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg
2581 ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac gggagtccaa
2641 gcgagctcgg tactaaaaca attcatccag taaaatataa tattttattt tctcccaatc
2701 aggcttgatc cccagtaagt caaaaaatag ctcgacatac tgttcttccc cgatatcctc
2761 cctgatcgac cggacgcaga aggcaatgtc ataccacttg tccgccctgc cgcttctccc
2821 aagatcaata aagccactta ctttgccatc tttcacaaag atgttgctgt ctcccaggtc
2881 gccgtgggaa aagacaagtt cctcttcggg cttttccgtc tttaaaaaat catacagctc
2941 gcgcggatct ttaaatggag tgtcttcttc ccagttttcg caatccacat cggccagatc
3001 gttattcagt aagtaatcca attcggctaa gcggccgtct aagctattcg tatagggaca
3061 atccgatatg tcgatggagt gaaagagcct gatgcactcc gcatacagct cgatagtctt
3121 ttcagggctt tgttcatctt catacccttc cgagcaaagg acgccatcgg cctcactcat
3181 gagcagattg ctccagccat catgccgttc aaagtgcagg acctttggaa caggcagctt
3241 tccttccagc catagcatca tgtccttttc ccgttccaca tcataggtgg tccctttata
3301 ccggctgtcc gtcattttta aatataggat ttcattttct cccaccagct tatatacctt
3361 agcaggagac attccttccg tatcttttac gcagcggtat tcttcgatca gttttttcaa
3421 ttccggtgat attctcattt tagccattta ttatttcctt cctcttttct acagtattta
3481 aagatacccc aagaagctaa ttataacaag acgaactcca attcactgtt ccttgcattc
3541 taaaacctta aatacagaaa acagcctttt caaagttgtt ttcaaagttg gcgtataaca
3601 tagtatcgac ggagccgatt ttgaaaccac aattatgata gaatttgacg tcg
//
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