| 出品公司: | -- |
|---|---|
| 载体名称: | pPZP212 |
| 质粒类型: | 植物载体；农杆菌双元表达载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 克隆方法: | 限制性内切酶，多克隆位点 |
| 启动子: | lac |
| 载体大小: | 9105 bp |
| 5' 测序引物及序列: | M13 Reverse：CAGGAAACAGCTATGAC |
| 3' 测序引物及序列: | M13/pUC Forward：CCCAGTCACGACGTTGTAAAACG |
| 载体标签: | 无 |
| 载体抗性: | 卡那霉素、壮观霉素(链霉素) |
| 筛选标记: | -- |
| 克隆菌株: | HB101 |
| 宿主细胞（系）: | 植物细胞、农杆菌 |
| 备注: | 重组质粒可用蓝白斑法筛选 |
| 产品目录号: | -- |
| 稳定性: | 稳表达 |
| 组成型/诱导型: | 诱导型 |
| 病毒/非病毒: | 非病毒 |
LOCUS Exported 9015 bp ds-DNA circular SYN 28-JUL-2016
DEFINITION Agrobacterium binary vector for plant transformation, with
spectinomycin- and kanamycin-resistance genes. The MCS is reversed
in pPZP211.
ACCESSION U10462
VERSION .
KEYWORDS pPZP212
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9015)
AUTHORS Hajdukiewicz P, Svab Z, Maliga P.
TITLE The small, versatile pPZP family of Agrobacterium binary vectors for
plant transformation.
JOURNAL Plant Mol. Biol. 1994;25:989-94.
PUBMED 7919218
REFERENCE 2 (bases 1 to 9015)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Thursday, July 28, 2016 from SnapGene Viewer 3.1.4
http://www.snapgene.com
COMMENT The GenBank sequence was corrected by inserting a G at position
2249.
FEATURES Location/Qualifiers
source 1..9015
/organism="synthetic DNA construct"
/lab_host="Plant Cells"
/mol_type="other DNA"
CDS 1187..1816
/codon_start=1
/product="stability protein from Pseudomonas plasmid pVS1"
/note="pVS1 StaA"
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 2245..3318
/codon_start=1
/product="replication protein from Pseudomonas plasmid
pVS1"
/note="pVS1 RepA"
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
rep_origin 3384..3578
/note="pVS1 oriV"
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 3922..4062
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(4248..4836)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5080..5871)
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase"
/note="SmR"
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
misc_feature 6399..6423
/note="LB T-DNA repeat"
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal 6521..6695
/note="CaMV poly(A) signal"
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(6749..7546)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
promoter complement(7606..8284)
/note="CaMV 35S promoter (enhanced)"
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
CDS complement(8370..8603)
/codon_start=1
/gene="lacZ (truncated)"
/product="LacZ-alpha fragment of beta-galactosidase"
/note="lacZ-alpha"
/translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
LAAHPPFASWRNSEEARTDRPSQQLRSLNGEC"
primer_bind 8513..8529
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 8533..8589
/note="MCS"
/note="pUC18 multiple cloning site"
primer_bind complement(8599..8615)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature complement(8601..8603)
/note="ATG"
protein_bind 8623..8639
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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(8647..8677)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
misc_feature 8878..8902
/note="RB T-DNA repeat"
/note="right border repeat from nopaline C58 T-DNA"
ORIGIN
1 agtactttga tccaacccct ccgctgctat agtgcagtcg gcttctgacg ttcagtgcag
61 ccgtcttctg aaaacgacat gtcgcacaag tcctaagtta cgcgacaggc tgccgccctg
121 cccttttcct ggcgttttct tgtcgcgtgt tttagtcgca taaagtagaa tacttgcgac
181 tagaaccgga gacattacgc catgaacaag agcgccgccg ctggcctgct gggctatgcc
241 cgcgtcagca ccgacgacca ggacttgacc aaccaacggg ccgaactgca cgcggccggc
301 tgcaccaagc tgttttccga gaagatcacc ggcaccaggc gcgaccgccc ggagctggcc
361 aggatgcttg accacctacg ccctggcgac gttgtgacag tgaccaggct agaccgcctg
421 gcccgcagca cccgcgacct actggacatt gccgagcgca tccaggaggc cggcgcgggc
481 ctgcgtagcc tggcagagcc gtgggccgac accaccacgc cggccggccg catggtgttg
541 accgtgttcg ccggcattgc cgagttcgag cgttccctaa tcatcgaccg cacccggagc
601 gggcgcgagg ccgccaaggc ccgaggcgtg aagtttggcc cccgccctac cctcaccccg
661 gcacagatcg cgcacgcccg cgagctgatc gaccaggaag gccgcaccgt gaaagaggcg
721 gctgcactgc ttggcgtgca tcgctcgacc ctgtaccgcg cacttgagcg cagcgaggaa
781 gtgacgccca ccgaggccag gcggcgcggt gccttccgtg aggacgcatt gaccgaggcc
841 gacgccctgg cggccgccga gaatgaacgc caagaggaac aagcatgaaa ccgcaccagg
901 acggccagga cgaaccgttt ttcattaccg aagagatcga ggcggagatg atcgcggccg
961 ggtacgtgtt cgagccgccc gcgcacgtct caaccgtgcg gctgcatgaa atcctggccg
1021 gtttgtctga tgccaagctg gcggcctggc cggccagctt ggccgctgaa gaaaccgagc
1081 gccgccgtct aaaaaggtga tgtgtatttg agtaaaacag cttgcgtcat gcggtcgctg
1141 cgtatatgat gcgatgagta aataaacaaa tacgcaaggg gaacgcatga aggttatcgc
1201 tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc gcaacccatc tagcccgcgc
1261 cctgcaactc gccggggccg atgttctgtt agtcgattcc gatccccagg gcagtgcccg
1321 cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt gtcggcatcg accgcccgac
1381 gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc gtagtgatcg acggagcgcc
1441 ccaggcggcg gacttggctg tgtccgcgat caaggcagcc gacttcgtgc tgattccggt
1501 gcagccaagc ccttacgaca tatgggccac cgccgacctg gtggagctgg ttaagcagcg
1561 cattgaggtc acggatggaa ggctacaagc ggcctttgtc gtgtcgcggg cgatcaaagg
1621 cacgcgcatc ggcggtgagg ttgccgaggc gctggccggg tacgagctgc ccattcttga
1681 gtcccgtatc acgcagcgcg tgagctaccc aggcactgcc gccgccggca caaccgttct
1741 tgaatcagaa cccgagggcg acgctgcccg cgaggtccag gcgctggccg ctgaaattaa
1801 atcaaaactc atttgagtta atgaggtaaa gagaaaatga gcaaaagcac aaacacgcta
1861 agtgccggcc gtccgagcgc acgcagcagc aaggctgcaa cgttggccag cctggcagac
1921 acgccagcca tgaagcgggt caactttcag ttgccggcgg aggatcacac caagctgaag
1981 atgtacgcgg tacgccaagg caagaccatt accgagctgc tatctgaata catcgcgcag
2041 ctaccagagt aaatgagcaa atgaataaat gagtagatga attttagcgg ctaaaggagg
2101 cggcatggaa aatcaagaac aaccaggcac cgacgccgtg gaatgcccca tgtgtggagg
2161 aacgggcggt tggccaggcg taagcggctg ggttgtctgc cggccctgca atggcactgg
2221 aacccccaag cccgaggaat cggcgtgagc ggtcgcaaac catccggccc ggtacaaatc
2281 ggcgcggcgc tgggtgatga cctggtggag aagttgaagg ccgcgcaggc cgcccagcgg
2341 caacgcatcg aggcagaagc acgccccggt gaatcgtggc aagcggccgc tgatcgaatc
2401 cgcaaagaat cccggcaacc gccggcagcc ggtgcgccgt cgattaggaa gccgcccaag
2461 ggcgacgagc aaccagattt tttcgttccg atgctctatg acgtgggcac ccgcgatagt
2521 cgcagcatca tggacgtggc cgttttccgt ctgtcgaagc gtgaccgacg agctggcgag
2581 gtgatccgct acgagcttcc agacgggcac gtagaggttt ccgcagggcc ggccggcatg
2641 gccagtgtgt gggattacga cctggtactg atggcggttt cccatctaac cgaatccatg
2701 aaccgatacc gggaagggaa gggagacaag cccggccgcg tgttccgtcc acacgttgcg
2761 gacgtactca agttctgccg gcgagccgat ggcggaaagc agaaagacga cctggtagaa
2821 acctgcattc ggttaaacac cacgcacgtt gccatgcagc gtacgaagaa ggccaagaac
2881 ggccgcctgg tgacggtatc cgagggtgaa gccttgatta gccgctacaa gatcgtaaag
2941 agcgaaaccg ggcggccgga gtacatcgag atcgagctag ctgattggat gtaccgcgag
3001 atcacagaag gcaagaaccc ggacgtgctg acggttcacc ccgattactt tttgatcgat
3061 cccggcatcg gccgttttct ctaccgcctg gcacgccgcg ccgcaggcaa ggcagaagcc
3121 agatggttgt tcaagacgat ctacgaacgc agtggcagcg ccggagagtt caagaagttc
3181 tgtttcaccg tgcgcaagct gatcgggtca aatgacctgc cggagtacga tttgaaggag
3241 gaggcggggc aggctggccc gatcctagtc atgcgctacc gcaacctgat cgagggcgaa
3301 gcatccgccg gttcctaatg tacggagcag atgctagggc aaattgccct agcaggggaa
3361 aaaggtcgaa aaggtctctt tcctgtggat agcacgtaca ttgggaaccc aaagccgtac
3421 attgggaacc ggaacccgta cattgggaac ccaaagccgt acattgggaa ccggtcacac
3481 atgtaagtga ctgatataaa agagaaaaaa ggcgattttt ccgcctaaaa ctctttaaaa
3541 cttattaaaa ctcttaaaac ccgcctggcc tgtgcataac tgtctggcca gcgcacagcc
3601 gaagagctgc aaaaagcgcc tacccttcgg tcgctgcgct ccctacgccc cgccgcttcg
3661 cgtcggccta tcgcggccgc tggccgctca aaaatggctg gcctacggcc aggcaatcta
3721 ccagggcgcg gacaagccgc gccgtcgcca ctcgaccgcc ggcgcccaca tcaaggcacc
3781 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
3841 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
3901 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
3961 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
4021 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
4081 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
4141 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
4201 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
4261 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
4321 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
4381 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
4441 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
4501 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
4561 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
4621 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
4681 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
4741 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
4801 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
4861 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gcatgatata
4921 tctcccaatt tgtgtagggc ttattatgca cgcttaaaaa taataaaagc agacttgacc
4981 tgatagtttg gctgtgagca attatgtgct tagtgcatct aatcgcttga gttaacgccg
5041 gcgaagcggc gtcggcttga acgaatttct agctagacat tatttgccga ctaccttggt
5101 gatctcgcct ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg aggccaagcg
5161 atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc
5221 cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac
5281 tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc
5341 gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg
5401 aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc
5461 ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag
5521 aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg
5581 aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc
5641 aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag
5701 ccttacggtc accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatccac
5761 tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc
5821 aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccccca tgatgtttaa
5881 ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctccata acatcaaaca
5941 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtaccccaaa
6001 aaaacatgtc ataacaagaa gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt
6061 cggtcaaggt tctggaccag ttgcgtgacg gcagttacgc tacttgcatt acagcttacg
6121 aaccgaacga ggcttatgtc cactgggttc gtgcccgaat tgatcacagg cagcaacgct
6181 ctgtcatcgt tacaatcaac atgctaccct ccgcgagatc atccgtgttt caaacccggc
6241 agcttagttg ccgttcttcc gaatagcatc ggtaacatga gcaaagtctg ccgccttaca
6301 acggctctcc cgctgacgcc gtcccggact gatgggctgc ctgtatcgag tggtgatttt
6361 gtgccgagct gccggtcggg gagctgttgg ctggctggtg gcaggatata ttgtggtgta
6421 aacaaattga cgcttagaca acttaataac acattgcgga cgtttttaat gtactgaatt
6481 aacgccgaat taattcgggg gatctatagg gactttaggt gatctggatt ttagtactgg
6541 attttggttt taggaattag aaattttatt gatagaagta ttttacaaat acaaatacat
6601 actaagggtt tcttatatgc tcaacacgtg agcgaaaccc tataagaacc ctaattccct
6661 tatctgggaa ctactcacac attattatgg agaaactcga tgtcgatcga ctctagctag
6721 aggatcgatc cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg
6781 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt
6841 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg
6901 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat
6961 tcggcaagca ggcatcgcca tgtgtcacga cgagatcctc gccgtcgggc atgcgcgcct
7021 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct
7081 gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt
7141 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga
7201 tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc
7261 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa
7321 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ctggagttca ttcagggcac
7381 cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg
7441 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc
7501 aagcggccgg agaacctgcg tgcaatccat cttgttcaat ccccatggtc gatcgacaga
7561 tctgcgaatc gagagagata gatttgtaga gagagactgg tgatttcagc gtgtcctctc
7621 caaatgaaat gaacttcctt atatagagga agggtcttgc gaaggatagt gggattgtgc
7681 gtcatccctt acgtcagtgg agatatcaca tcaatccact tgctttgaag acgtggttgg
7741 aacgtcttct ttttccacga tgctcctcgt gggtgggggt ccatctttgg gaccactgtc
7801 ggcagaggca tcttgaacga tagcctttcc tttatcgcaa tgatggcatt tgtaggtgcc
7861 accttccttt tctactgtcc ttttgatgaa gtgacagata gctgggcaat ggaatccgag
7921 gaggtttccc gatattaccc tttgttgaaa agtctcaata gccctttggt cttctgagac
7981 tgtatctttg atattcttgg agtagacgag agtgtcgtgc tccaccatgt tatcacatca
8041 atccacttgc tttgaagacg tggttggaac gtcttctttt tccacgatgc tcctcgtggg
8101 tgggggtcca tctttgggac cactgtcggc agaggcatct tgaacgatag cctttccttt
8161 atcgcaatga tggcatttgt aggtgccacc ttccttttct actgtccttt tgatgaagtg
8221 acagatagct gggcaatgga atccgaggag gtttcccgat attacccttt gttgaaaagt
8281 ctcaatagcc ctttggtctt ctgagactgt atctttgata ttcttggagt agacgagagt
8341 gtcgtgctcc accatgttgg caagctgctc tagcattcgc cattcaggct gcgcaactgt
8401 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
8461 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
8521 acggccagtg ccaagcttgc atgcctgcag gtcgactcta gaggatcccc gggtaccgag
8581 ctcgaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
8641 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
8701 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
8761 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttggctagag
8821 cagcttgagc ttggatcaga ttgtcgtttc ccgccttcag tttaaactat cagtgtttga
8881 caggatatat tggcgggtaa acctaagaga aaagagcgtt tattagaata acggatattt
8941 aaaagggcgt gaaaaggttt atccgttcgt ccatttgtat gtgcatgcca accacagggt
9001 tcccctcggg atcaa
//
