| 出品公司: | -- |
|---|---|
| 载体名称: | pGreenII0049 ; pGreenII 0049 |
| 质粒类型: | 植物载体；农杆菌双元表达载体；荧光素酶报告载体 |
| 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: | 7065 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: | 荧光素酶 |
| 载体抗性: | 卡那霉素 |
| 筛选标记: | Neomycin |
| 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 7065 bp ds-DNA circular SYN 14-七月-2016
DEFINITION Agrobacterium binary vector with kanamycin-resistance and luciferase
genes.
ACCESSION .
VERSION .
KEYWORDS pGreenII 0049
SOURCE http://www.biofeng.com/
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7065)
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 7065)
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..7065
/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="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(970..1764)
/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))"
/note="color: #ccffcc"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERGRTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTQGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
promoter complement(1798..1977)
/note="NOS promoter"
/note="nopaline synthase promoter"
/note="color: #000000; direction: LEFT"
CDS complement(2048..2437)
/codon_start=1
/gene="lacZ (truncated)"
/product="LacZ-alpha fragment of?beta-galactosidase"
/note="lacZ"
/note="color: #993366"
/translation="MTMITPSSELTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
PSTSRGGPVPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
DRPSQQLRSLNGEWREIVNVNVIDT"
primer_bind 2224..2240
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
/note="color: #a020f0; direction: RIGHT"
promoter 2247..2265
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
/note="color: #ffffff; direction: RIGHT"
misc_feature 2274..2381
/note="MCS"
/note="pBluescript multiple cloning site"
/note="color: #99ccff"
promoter complement(2394..2412)
/note="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
/note="color: #ffffff; direction: LEFT"
primer_bind complement(2433..2449)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
/note="color: #a020f0; direction: LEFT"
protein_bind 2457..2473
/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(2481..2512)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
/note="This reverse directional feature has 3 segments:
???1:?2481?..?2487?/?#ffffff?/?-10
???2:?2488?..?2506?/?#ffffff
???3:?2507?..?2512?/?#ffffff?/?-35"
polyA_signal 2787..2963
/note="CaMV poly(A)"
/note="cauliflower mosaic virus polyadenylation signal"
/note="color: #a6acb3"
CDS complement(3079..4731)
/codon_start=1
/gene="luc"
/product="firefly luciferase"
/EC_number="
"
/note="luciferase"
/note="color: #ffcc00"
/experiment="
"
/protein_id="
"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
promoter complement(4742..5087)
/note="CaMV 35S promoter"
/note="strong constitutive promoter from cauliflower mosaic
virus"
/note="color: #000000; direction: LEFT"
misc_feature 5148..5172
/note="RB"
/note="right border repeat from nopaline C58 T-DNA"
/note="color: #ffe4c4"
rep_origin complement(5263..5851)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/note="color: #ffff00"
CDS complement(6022..6837)
/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 tagctagagt ccgaacccca
961 gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg
1021 agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc
1081 aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca
1141 gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc
1201 ctgggtcacg acgagatcct cgccgtcggg catccgcgcc ttgagcctgg cgaacagttc
1261 ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc
1321 catccgagta cgtcctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc
1381 cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg
1441 agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct
1501 tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca
1561 cgatagccgc gctgcctcgt cttggagttc attcagggca ccggacaggt cggtcttgac
1621 aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat
1681 tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc
1741 gtgcaatcca tcttgttcaa tcatgcctcg atcgagttga gagtgaatat gagactctaa
1801 ttggataccg aggggaattt atggaacgtc agtggagcat ttttgacaag aaatatttgc
1861 tagctgatag tgaccttagg cgacttttga acgcgcaata atggtttctg acgtatgtgc
1921 ttagctcatt aaactccaga aacccgcggc tgagtggctc cttcaacgtt gcggttctgt
1981 cagttccaaa cgtaaaacgg cttgtcccgc gtcatcggcg ggggtcataa cgtgactccc
2041 ttaattctca tgtatcgata acattaacgt ttacaatttc gcgccattcg ccattcaggc
2101 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
2161 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
2221 gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa ttgggtaccg
2281 ggccccccct cgaggtcgac ggtatcgata agcttgatat cgaattcctg cagcccgggg
2341 gatccactag ttctagagcg gccgccaccg cggtggagct ccagcttttg ttccctttag
2401 tgagggttaa ttccgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
2461 tatccgctca caattccaca caacatacga gccggaaghc ataaagtgta aagcctgggg
2521 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
2581 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
2641 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
2701 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
2761 taacgcagga aagaacatga aggatcgatc tggattttag tactggattt tggttttagg
2821 aattagaaat tttattgata gaagtatttt acaaatacaa atacatacta agggtttctt
2881 atatgctcaa cacatgagcg aaaccctata agaaccctaa tttcccttat cgggaaacta
2941 ctcacacatt atttatggag aaaatagaga gagatagatt tgtagagaga gactggtgat
3001 ttcagcgtac cgaattaatt ctctagctag agtattacaa tagctaagaa tttcgtcatc
3061 gctgaataca gttacatttt acaatttgga ctttccgccc ttcttggcct ttatgaggat
3121 ctctctgatt tttcttgcgt cgagttttcc ggtaagacct ttcggtactt cgtccacaaa
3181 cacaactcct ccgcgcaact ttttggcggt tgttacttga ctggcgacgt aatccacgat
3241 ctctttttcc gtcatcgtct ttccgtgctc caaaacaaca acggcggcgg gaagttcacc
3301 ggcgtcatcg tcgggaagac ctgccacgcc cgcgtcgaag atgttggggt gttgtaacaa
3361 tatcgactcc aattcagcgg gggccacctg gtatcctttg tatttaatta aagacttcaa
3421 gcggtcaact atgaagaagt gttcgtcttc gtcccagtaa gctatgtctc cagaatgtag
3481 ccatccatcc ttgtcaatca aggcgttggt cgcttccgga ttgtttacat aaccggacat
3541 aatcataggt cctctgacac ataattcgcc tctctgatta acgcccagcg ttttcccggt
3601 atccagatcc acaaccttcg cttcaaaaaa tggaacaact ttaccgaccg cgcccggttt
3661 atcatccccc tcgggtgtaa tcagaatagc tgatgtagtc tcagtgagcc catatccttg
3721 tcgtatccct ggaagatgga agcgttttgc aaccgcttcc ccgacttctt tcgaaagagg
3781 tgcgccccca gaagcaattt cgtgtaaatt agataaatcg tatttgtcaa tcagagtgct
3841 tttggcgaag aatgaaaata gggttggtac tagcaacgca ctttgaattt tgtaatcctg
3901 aagggatcgt aaaaacagct cttcttcaaa tctatacatt aagacgactc gaaatccaca
3961 tatcaaatat ccgagtgtag taaacattcc aaaaccgtga tggaatggaa caacacttaa
4021 aatcgcagta tccggaatga tttgattgcc aaaaatagga tctctggcat gcgagaatct
4081 gacgcaggca gttctatgcg gaagggccac acccttaggt aacccagtag atccagagga
4141 gttcattatc agtgcaattg ttttgtcacg atcaaaggac tctggtacaa aatcgtattc
4201 attaaaaccg ggaggtagat gagatgtgac gaacgtgtac atcgactgaa atccctggta
4261 atccgtttta gaatccatga taataatttt ctggattatt ggtaattttt tttgcacgtt
4321 caaaattttt tgcaacccct ttttggaaac aaacactacg gtaggctgcg aaatgttcat
4381 actgttgagc aattcacgtt cattataaat gtcgttcgcg ggcgcaactg caactccgat
4441 aaataacgcg cccaacaccg gcataaagaa ttgaagagag ttttcactgc atacgacgat
4501 tctgtgattt gtattcagcc catatcgttt catagcttct gccaaccgaa cggacatttc
4561 gaagtattcc gcgtacgtga tgttcacctc gatatgtgca tctgtaaaag caattgttcc
4621 aggaaccagg gcgtatctct tcatagcctt atgcagttgc tctccagcgg ttccatcctc
4681 tagcggatag aatggcgccg ggcctttctt tatgtttttg gcgtcttcca tgagcttggg
4741 ctgtcctctc caaatgaaat gaacttcctt atatagagga agggtcttgc gaaggatagt
4801 gggattgtgc gtcatccctt acgtcagtgg agatgtcaca tcaatccact tgctttgaag
4861 acgtggttgg aacgtcttct ttttccacga tgctcctcgt gggtgggggt ccatctttgg
4921 gaccactgtc ggcagaggca tcttgaatga tagcctttcc tttatcgcaa tgatggcatt
4981 tgtaggagcc accttccttt tctactgtcc tttcgatgaa gtgacagata gctgggcaat
5041 ggaatccgag gaggtttccc gaaattaccc tttgttgaaa agtctcaata gccctttggt
5101 cttctgagac tgtatctttg acatttttgg agtaggggta cgatccttga caggatatat
5161 tggcgggtaa actaagtcgc tgtatgtgtt tgtttgagat ctcatgtgag caaaaggcca
5221 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
5281 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
5341 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
5401 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
5461 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
5521 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
5581 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
5641 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
5701 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa gaagagttgg
5761 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
5821 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
5881 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
5941 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
6001 tgtgtaacat tggtctagtg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt
6061 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
6121 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
6181 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
6241 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
6301 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
6361 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
6421 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
6481 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc ctgggatcgc
6541 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
6601 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacaacat tggcaacgct
6661 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcggtagat
6721 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
6781 catgttggaa tttaatcgcg gccttgagca agacgtttcc cgttgaatat ggctcataac
6841 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt
6901 atcttgtgca atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg
6961 aacttttgct gagttgaagg atcagatcac gcatcttccc gacaacgcag accgttccgt
7021 ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac aacaa
//
