| 出品公司: | -- |
|---|---|
| 载体名称: | pGreenII0229 ; pGreen II0229 |
| 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: | lacZ |
| 载体大小: | 4448 bp |
| 5' 测序引物及序列: | M13 Forward：TGTAAAACGACGGCCAGT |
| 3' 测序引物及序列: | M13 Reverse：CAGGAAACAGCTATGAC |
| 载体标签: | -- |
| 载体抗性: | 卡那霉素 |
| 筛选标记: | 双丙胺膦/草丁膦(bialophos / phosphinothricin) |
| 克隆菌株: | HB101等菌株 |
| 宿主细胞（系）: | 农杆菌/植物细胞 |
| 备注: | -- |
| 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 File 4448 bp ds-DNA circular SYN 14-七月-2016
DEFINITION Agrobacterium binary vector with kanamycin-and
bialophos/phosphinothricin-resistance genes.
ACCESSION .
VERSION .
KEYWORDS pGreenII 0229
SOURCE http://www.biofeng.com/
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4448)
AUTHORS Hellens RP, Edwards EA, Leyland NR, Bean S, Mullineaux PM.
TITLE pGreen: a versatile and flexible binary Ti vector for
Agrobacterium-mediated plant transformation.
JOURNAL Plant Mol. Biol. 2000;42:819-32.
PUBMED 10890530
REFERENCE 2 (bases 1 to 4448)
AUTHORS pGreen website
TITLE Direct Submission
JOURNAL Exported 2016-07-14 from SnapGene Viewer 1.5.3
http://www.snapgene.com
COMMENT This plasmid must be transformed into an Agrobacterium strain that
also carries a vector such as pSoup.
FEATURES Location/Qualifiers
source 1..4448
/organism="synthetic DNA construct"
/lab_host="Plant Cells"
/mol_type="other DNA"
rep_origin 63..498
/gene="
"
/note="pSa ori"
/note="origin of replication from bacterial plasmid pSa"
/note="color: #ffff00"
misc_feature 633..655
/note="LB"
/note="LB T-DNA repeat ?left border repeat from nopaline
C58 T-DNA (truncated)"
/note="color: #ffe4c4"
terminator 679..930
/note="NOS terminator"
/note="nopaline synthase terminator and poly(A) signal"
/note="color: #ffffff"
CDS complement(950..1501)
/codon_start=1
/gene="Streptomyces hygroscopicus bar"
/product="phosphinothricin acetyltransferase"
/function="
"
/note="BlpR"
/note="confers resistance to bialophos or phosphinothricin"
/note="color: #ccffcc"
/protein_id="
"
/translation="MSPERRPADIRRATEADMPAVCTIVNHYIQTSTVNFRTEPQEPQE
WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
WQLDFSLPVPPRPVLPVTEI"
promoter complement(1544..1721)
/note="NOS promoter"
/note="nopaline synthase promoter"
/note="color: #ffffff; direction: LEFT"
CDS complement(1792..2181)
/codon_start=1
/gene="lacZ (truncated)"
/product="LacZ-alpha fragment of?beta-galactosidase"
/note="lacZ"
/note="color: #993366"
/translation="MTMITPSSELTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
PSTSRGGPVPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
DRPSQQLRSLNGEWREIVNVNVIDT"
primer_bind 1968..1984
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
/note="color: #a020f0; direction: RIGHT"
promoter 1991..2009
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
/note="color: #ffffff; direction: RIGHT"
misc_feature 2018..2125
/note="MCS"
/note="pBluescript multiple cloning site"
/note="color: #00ff00"
promoter complement(2138..2156)
/note="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
/note="color: #ffffff; direction: LEFT"
primer_bind complement(2177..2193)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
/note="color: #a020f0; direction: LEFT"
protein_bind 2201..2217
/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)."
/note="color: #31849b"
promoter complement(2225..2256)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
/note="This reverse directional feature has 3 segments:
???1:?2225?..?2231?/?#ffffff?/?-10
???2:?2232?..?2250?/?#ffffff
???3:?2251?..?2256?/?#ffffff?/?-35"
misc_feature 2531..2555
/note="RB"
/note="RB T-DNA repeat ?right border repeat from nopaline
C58 T-DNA"
/note="color: #ffe4c4"
rep_origin complement(2646..3234)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/note="color: #ffff00"
CDS complement(3405..4220)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/note="KanR"
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/note="color: #ccffcc"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANVVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
1 agctctcatc aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggcgat
61 ccccatccaa cagcccgccg tcgagcgggc ttttttatcc ccggaagcct gtggatagag
121 ggtagttatc cacgtgaaac cgctaatgcc ccgcaaagcc ttgattcacg gggctttccg
181 gcccgctcca aaaactatcc acgtgaaatc gctaatcagg gtacgtgaaa tcgctaatcg
241 gagtacgtga aatcgctaat aaggtcacgt gaaatcgcta atcaaaaagg cacgtgagaa
301 cgctaatagc cctttcagat caacagcttg caaacacccc tcgctccggc aagtagttac
361 agcaagtagt atgttcaatt agcttttcaa ttatgaatat atatatcaat tattggtcgc
421 ccttggcttg tggacaatgc gctacgcgca ccggctccgc ccgtggacaa ccgcaagcgg
481 ttgcccaccg tcgagcgcca gcgcctttgc ccacaacccg gcggccggcc gcaacagatc
541 gttttataaa tttttttttt tgaaaaagaa aaagcccgaa aggcggcaac ctctcgggct
601 tctggatttc cgatccccgg aattagagat cttggcagga tatattgtgg tgtaacgtta
661 tcagcttgca tgccggtcga tctagtaaca tagatgacac cgcgcgcgat aatttatcct
721 agtttgcgcg ctatattttg ttttctatcg cgtattaaat gtataattgc gggactctaa
781 tcaaaaaacc catctcataa ataacgtcat gcattacatg ttaattatta catgcttaac
841 gtaattcaac agaaattata tgataatcat cgcaagaccg gcaacaggat tcaatcttaa
901 gaaactttat tgccaaatgt ttgaacgatc tgcttgactc taggggtcat cagatttcgg
961 tgacgggcag gaccggacgg ggcggcaccg gcaggctgaa gtccagctgc cagaaaccca
1021 cgtcatgcca gttcccgtgc ttgaagccgg ccgcccgcag catgccacgg ggggcatatc
1081 cgagcgcctc gtgcatgcgc acgctcgggt cgttgggcag cccgatgaca gcgaccacgc
1141 tcttgaagcc ctgtgcctcc agggacttca gcaggtgggt gtagagcgtg gagcccagtc
1201 ccgtccgctg gtggcggggg gagacgtaca cggttgactc ggccgtccag tcgtaggcgt
1261 tgcgtgcctt ccagggaccc gcgtaggcga tgccggcgac ctcgccgtcc acctcggcga
1321 cgagccaggg atagcgctcc cgcagacgga cgaggtcgtc cgtccactcc tgcggttcct
1381 gcggctcggt acggaagttg accgtgcttg tctggatgta gtggttgacg atggtgcaga
1441 ccgccggcat gtccgcctcg gtggcacggc ggatgtcggc cgggcgtcgt tctgggctca
1501 tggtagatcc ccctcgatcg agttgagagt gaatatgaga ctctaattgg ataccgaggg
1561 gaatttatgg aacgtcagtg gagcattttt gacaagaaat atttgctagc tgatagtgac
1621 cttaggcgac ttttgaacgc gcaataatgg tttctgacgt atgtgcttag ctcattaaac
1681 tccagaaacc cggctgagtg gctccttcaa cgttgcggtt ctgtcagttc caaacgtaaa
1741 acggcttgtc ccgcgtcatc ggcgggggtc ataacgtgac tcccttaatt ctcatgtatc
1801 gataacatta acgtttacaa tttcgcgcca ttcgccattc aggctgcgca actgttggga
1861 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
1921 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
1981 cagtgaattg taatacgact cactataggg cgaattgggt accgggcccc ccctcgaggt
2041 cgacggtatc gataagcttg atatcgaatt cctgcagccc gggggatcca ctagttctag
2101 agcggccgcc accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattccga
2161 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
2221 cacacaacat acgagccgga aghcataaag tgtaaagcct ggggtgccta atgagtgagc
2281 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
2341 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
2401 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
2461 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
2521 atgaaggcct tgacaggata tattggcggg taaactaagt cgctgtatgt gtttgtttga
2581 gatctcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
2641 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
2701 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
2761 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
2821 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
2881 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
2941 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
3001 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
3061 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
3121 gttaccttcg gaagaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
3181 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
3241 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
3301 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
3361 tttaaatcaa tctaaagtat atatgtgtaa cattggtcta gtgattagaa aaactcatcg
3421 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
3481 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
3541 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
3601 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
3661 ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
3721 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
3781 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
3841 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
3901 aatgctgttt tccctgggat cgcagtggtg agtaaccatg catcatcagg agtacggata
3961 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
4021 tctgtaacaa cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
4081 ggcttcccat acaatcggta gattgtcgca cctgattgcc cgacattatc gcgagcccat
4141 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggccttga gcaagacgtt
4201 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
4261 attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca
4321 acgtggcttt gttgaataaa tcgaactttt gctgagttga aggatcagat cacgcatctt
4381 cccgacaacg cagaccgttc cgtggcaaag caaaagttca aaatcaccaa ctggtccacc
4441 tacaacaa
