| 质粒名称: | pPMQAK1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 26052 |
| 存储实验室: | Peter Lindblad |
|
载体骨架基本信息 |
|
| 载体名称: | RSF1010 derivative pAWG1.1 + pSB1AK3-BBa_B0015 |
| 空载体大小: | 8372bp |
| 完整质粒大小: | 8372 bp |
| 载体类型: | 细菌细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | Ampicillin and Kanamycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | DB3.1 |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | BioBrick-compatible cyanobacterial shuttle vector |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | see comments section(未破坏) |
| 3’克隆位点: | see comments section (未破坏) |
| 5’测序引物 | Amp-R |
|
文献引用方法 |
|
| 材料和方法部分: |
pPMQAK1 was a gift from Peter Lindblad (Addgene plasmid # 26052)
|
| 参考文献部分: | Design and characterization of molecular tools for a Synthetic Biology approach towards developing cyanobacterial biotechnology. Huang HH, Camsund D, Lindblad P, Heidorn T. Nucleic Acids Res. 2010 May . 38(8):2577-93. 10.1093/nar/gkq164 PubMed 20236988 |
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 pPMQAK1 8372 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pPMQAK1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8372)
AUTHORS Huang HH, Camsund D, Lindblad P, Heidorn T
TITLE Design and characterization of molecular tools for a Synthetic
Biology approach towards developing cyanobacterial biotechnology.
JOURNAL Nucleic Acids Res. 2010 May . 38(8):2577-93.
PUBMED 20236988
REFERENCE 2 (bases 1 to 8372)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8372
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 576..595
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 660..679
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS complement(823..1683)
/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 1446..1465
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1684..1788)
/gene="bla"
/label=AmpR promoter
primer_bind 1856..1874
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
terminator 1878..1921
/label=bacterial terminator
/note="putative bacterial transcription terminator"
misc_feature 1924..1945
/label=BioBrick prefix
/note="BioBrick prefix for parts that do not start with
""ATG"""
CDS complement(1974..2279)
/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 complement(2076..2095)
/label=ccdB-fwd
/note="ccdB gene, forward primer"
misc_feature 2621..2641
/label=BioBrick suffix
/note="universal suffix for all parts"
terminator 2642..2699
/label=his operon terminator
/note="This putative transcriptin terminator from the E.
coli his operon has a 2-bp deletion introduced during
synthesis. Its efficiency has not been determined."
primer_bind complement(2723..2740)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin 2881..3275
/label=RSF1010 oriV
/note="replication origin of the broad-host-range plasmid
RSF1010; requires the RSF1010 RepA/B/C proteins for
replication (Scholz et al., 1989)"
oriT 3617..3704
/label=RSF1010 oriT
/note="origin of transfer of the broad-host-range plasmid
RSF1010 (Scholz et al., 1989)"
CDS 4943..5914
/codon_start=1
/gene="repB"
/product="replication protein B of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
/label=RSF1010 RepB
/translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
CDS 6425..7264
/codon_start=1
/gene="repA"
/product="replication protein A of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
/label=RSF1010 RepA
/translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
CDS complement(8225..668)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
1 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
61 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagctta
121 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
181 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg
241 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
301 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc
361 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
421 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
481 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
541 aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
601 aaatcagcat ccatgttgga atttaatcgc ggcctggagc aagacgtttc ccgttgaata
661 tggctcataa caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat
721 gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacaac gtggctttgt
781 tgaataaatc gaacttttgc tgagttgaag gatcagctcg agttaccaat gcttaatcag
841 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
901 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
961 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
1021 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
1081 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
1141 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
1201 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
1261 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
1321 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
1381 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
1441 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
1501 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
1561 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
1621 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
1681 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
1741 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
1801 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
1861 gcgtatcacg aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt
1921 ctggaattcg cggccgcttc tagagactgg ctgtgtataa gggagcctga catttatatt
1981 ccccagaaca tcaggttaat ggcgtttttg atgtcatttt cgcggtggct gagatcagcc
2041 acttcttccc cgataacgga gaccggcaca ctggccatat cggtggtcat catgcgccag
2101 ctttcatccc cgatatgcac caccgggtaa agttcacggg agactttatc tgacagcaga
2161 cgtgcactgg ccagggggat caccatccgt cgcccgggcg tgtcaataat atcactctgt
2221 acatccacaa acagacgata acggctctct cttttatagg tgtaaacctt aaactgcatt
2281 tcaccagccc ctgttctcgt cagcaaaaga gccgttcatt tcaataaacc gggcgacctc
2341 agccatccct tcctgatttt ccgctttcca gcgttcggca cgcagacgac gggcttcatt
2401 ctgcatggtt gcgcttacca gaccggagat attgacatca tatatgcctt gagcaactga
2461 tagctgtcgc tgtcaactgt cactgtaata cgctgcttca tagcatacct ctttttgaca
2521 tacttcgggt atacatatca gtatatattc ttataccgca aaaatcagcg cgcaaatacg
2581 catactgtta tctggctttt agtaagccgg atccacgcgt tactagtagc ggccgctgca
2641 gtccggcaaa aaagggcaag gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat
2701 tacttcgcgt tatgcaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
2761 ggcgagcggt atcagctcac tcaaaggcgg taatcaattg agttctttta ccctcagccg
2821 aaatgcctgc cgttgctaga cattgccagc cagtgcccgt cactcccgta ctaactgtca
2881 cgaacccctg caataactgt cacgcccccc tgcaataact gtcacgaacc cctgcaataa
2941 ctgtcacgcc cccaaacctg caaacccagc aggggcgggg gctggcgggg tgttggaaaa
3001 atccatccat gattatctaa gaataatcca ctaggcgcgg ttatcagcgc ccttgtgggg
3061 cgctgctgcc cttgcccaat atgcccggcc agaggccgga tagctggtct attcgctgcg
3121 ctaggctaca caccgcccca ccgctgcgcg gcagggggaa aggcgggcaa agcccgctaa
3181 accccacacc aaaccccgca gaaatacgct ggagcgcttt tagccgcttt agcggccttt
3241 ccccctaccc gaagggtggg ggcgcgtgtg cagccccgca gggcctgtct cggtcgatca
3301 ttcagcccgg ctcatccttc tggcgtggcg gcagaccgaa caaggcgcgg tcgtggtcgc
3361 gttcaaggta cgcatccatt gccgccatga gccgatcctc cggccactcg ctgctgttca
3421 ccttggccaa aatcatggcc cccaccagca ccttgcgcct tgtttcgttc ttgcgctctt
3481 gctgctgttc ccttgcccgc acccgctgaa tttcggcatt gattcgcgct cgttgttctt
3541 cgagcttggc cagccgatcc gccgccttgt tgctcccctt aaccatcttg acaccccatt
3601 gttaatgtgc tgtctcgtag gctatcatgg aggcacagcg gcggcaatcc cgaccctact
3661 ttgtagggga gggcgcactt accggtttct cttcgagaaa ctggcctaac ggccaccctt
3721 cgggcggtgc gctctccgag ggccattgca tggagccgaa aagcaaaagc aacagcgagg
3781 cagcatggcg atttatcacc ttacggcgaa aaccggcagc aggtcgggcg gccaatcggc
3841 cagggccaag gccgactaca tccagcgcga aggcaagtat gcccgcgaca tggatgaagt
3901 cttgcacgcc gaatccgggc acatgccgga gttcgtcgag cggcccgccg actactggga
3961 tgctgccgac ctgtatgaac gcgccaatgg gcggctgttc aaggaggtcg aatttgccct
4021 gccggtcgag ctgaccctcg accagcagaa ggcgctggcg tccgagttcg cccagcacct
4081 gaccggtgcc gagcgcctgc cgtatacgct ggccatccat gccggtggcg gcgagaaccc
4141 gcactgccac ctgatgatct ccgagcggat caatgacggc atcgagcggc ccgccgctca
4201 gtggttcaag cggtacaacg gcaagacccc ggagaagggc ggggcacaga agaccgaagc
4261 gctcaagccc aaggcatggc ttgagcagac ccgcgaggca tgggccgacc atgccaaccg
4321 ggcattagag cgggctggcc acgacgcccg cattgaccac agaacacttg aggcgcaggg
4381 catcgagcgc ctgcccggtg ttcacctggg gccgaacgtg gtggagatgg aaggccgggg
4441 catccgcacc gaccgggcag acgtggccct gaacatcgac accgccaacg cccagatcat
4501 cgacttacag gaataccggg aggcaataga ccatgaacgc aatcgacaga gtgaagaaat
4561 ccagaggcat caacgagtta gcggagcaga tcgaaccgct ggcccagagc atggcgacac
4621 tggccgacga agcccggcag gtcatgagcc agacccagca ggccagcgag gcgcaggcgg
4681 cggagtggct gaaagcccag cgccagacag gggcggcatg ggtggagctg gccaaagagt
4741 tgcgggaggt agccgccgag gtgagcagcg ccgcgcagag cgcccggagc gcgtcgcggg
4801 ggtggcactg gaagctatgg ctaaccgtga tgctggcttc catgatgcct acggtggtgc
4861 tgctgatcgc atcgttgctc ttgctcgacc tgacgccact gacaaccgag gacggctcga
4921 tctggctgcg cttggtggcc cgatgaagaa cgacaggact ttgcaggcca taggccgaca
4981 gctcaaggcc atgggctgtg agcgcttcga tatcggcgtc agggacgcca ccaccggcca
5041 gatgatgaac cgggaatggt cagccgccga agtgctccag aacacgccat ggctcaagcg
5101 gatgaatgcc cagggcaatg acgtgtatat caggcccgcc gagcaggagc ggcatggtct
5161 ggtgctggtg gacgacctca gcgagtttga cctggatgac atgaaagccg agggccggga
5221 gcctgccctg gtagtggaaa ccagcccgaa gaactatcag gcatgggtca aggtggccga
5281 cgccgcaggc ggtgaacttc gggggcagat tgcccggacg ctggccagcg agtacgacgc
5341 cgacccggcc agcgccgaca gccgccacta tggccgcttg gcgggcttca ccaaccgcaa
5401 ggacaagcac accacccgcg ccggttatca gccgtgggtg ctgctgcgtg aatccaaggg
5461 caagaccgcc accgctggcc cggcgctggt gcagcaggct ggccagcaga tcgagcaggc
5521 ccagcggcag caggagaagg cccgcaggct ggccagcctc gaactgcccg agcggcagct
5581 tagccgccac cggcgcacgg cgctggacga gtaccgcagc gagatggccg ggctggtcaa
5641 gcgcttcggt gatgacctca gcaagtgcga ctttatcgcc gcgcagaagc tggccagccg
5701 gggccgcagt gccgaggaaa tcggcaaggc catggccgag gccagcccag cgctggcaga
5761 gcgcaagccc ggccacgaag cggattacat cgagcgcacc gtcagcaagg tcatgggtct
5821 gcccagcgtc cagcttgcgc gggccgagct ggcacgggca ccggcacccc gccagcgagg
5881 catggacagg ggcgggccag atttcagcat gtagtgcttg cgttggtact cacgcctgtt
5941 atactatgag tactcacgca cagaaggggg ttttatggaa tacgaaaaaa gcgcttcagg
6001 gtcggtctac ctgatcaaaa gtgacaaggg ctattggttg cccggtggct ttggttatac
6061 gtcaaacaag gccgaggctg gccgcttttc agtcgctgat atggccagcc ttaaccttga
6121 cggctgcacc ttgtccttgt tccgcgaaga caagcctttc ggccccggca agtttctcgg
6181 tgactgatat gaaagaccaa aaggacaagc agaccggcga cctgctggcc agccctgacg
6241 ctgtacgcca agcgcgatat gccgagcgca tgaaggccaa agggatgcgt cagcgcaagt
6301 tctggctgac cgacgacgaa tacgaggcgc tgcgcgagtg cctggaagaa ctcagagcgg
6361 cgcagggcgg gggtagtgac cccgccagcg cctaaccacc aactgcctgc aaaggaggca
6421 atcaatggct acccataagc ctatcaatat tctggaggcg ttcgcagcag cgccgccacc
6481 gctggactac gttttgccca acatggtggc cggtacggtc ggggcgctgg tgtcgcccgg
6541 tggtgccggt aaatccatgc tggccctgca actggccgca cagattgcag gcgggccgga
6601 tctgctggag gtgggcgaac tgcccaccgg cccggtgatc tacctgcccg ccgaagaccc
6661 gcccaccgcc attcatcacc gcctgcacgc ccttggggcg cacctcagcg ccgaggaacg
6721 gcaagccgtg gctgacggcc tgctgatcca gccgctgatc ggcagcctgc ccaacatcat
6781 ggccccggag tggttcgacg gcctcaagcg cgccgccgag ggccgccgcc tgatggtgct
6841 ggacacgctg cgccggttcc acatcgagga agaaaacgcc agcggcccca tggcccaggt
6901 catcggtcgc atggaggcca tcgccgccga taccgggtgc tctatcgtgt tcctgcacca
6961 tgccagcaag ggcgcggcca tgatgggcgc aggcgaccag cagcaggcca gccggggcag
7021 ctcggtactg gtcgataaca tccgctggca gtcctacctg tcgagcatga ccagcgccga
7081 ggccgaggaa tggggtgtgg acgacgacca gcgccggttc ttcgtccgct tcggtgtgag
7141 caaggccaac tatggcgcac cgttcgctga tcggtggttc aggcggcatg acggcggggt
7201 gctcaagccc gccgtgctgg agaggcagcg caagagcaag ggggtgcccc gtggtgaagc
7261 ctaagaacaa gcacagcctc agccacgtcc ggcacgaccc ggcgcactgt ctggcccccg
7321 gcctgttccg tgccctcaag cggggcgagc gcaagcgcag caagctggac gtgacgtatg
7381 actacggcga cggcaagcgg atcgagttca gcggcccgga gccgctgggc gctgatgatc
7441 tgcgcatcct gcaagggctg gtggccatgg ctgggcctaa tggcctagtg cttggcccgg
7501 aacccaagac cgaaggcgga cggcagctcc ggctgttcct ggaacccaag tgggaggccg
7561 tcaccgctga tgccatggtg gtcaaaggta gctatcgggc gctggcaaag gaaatcgggg
7621 cagaggtcga tagtggtggg gcgctcaagc acatacagga ctgcatcgag cgcctttgga
7681 aggtatccat catcgcccag aatggccgca agcggcaggg gtttcggctg ctgtcggagt
7741 acgccagcga cgaggcggac gggcgcctgt acgtggccct gaaccccttg atcgcgcagg
7801 ccgtcatggg tggcggccag catgtgcgca tcagcatgga cgaggtgcgg gcgctggaca
7861 gcgaaaccgc ccgcctgctg caccagcggc tgtgtggctg gatcgacccc ggcaaaaccg
7921 gcaaggcttc catagatacc ttgtgcggct atgtctggcc gtcagaggcc agtggttcga
7981 ccatgcgcaa gcgccgccag cgggtgcgcg aggcgttgcc ggagctggtc gcgctgggct
8041 ggacggtaac cgagttcgcg gcgggcaagt acgacatcac ccggcccaag gcggcaggct
8101 gacccccccc actctattgt aaacaagaca tttttatctt ttatattcaa tggcttattt
8161 tcctgctaat tggtaatacc atggtaccgc tctgccagtg ttacaaccaa ttaaccaatt
8221 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
8281 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
8341 tccataggat ggcaagatcc tggtatcggt ct
//
