LOCUS Exported File 5138 bp ds-DNA circular SYN 21-四月-2013
DEFINITION Agrobacterium binary vector with kanamycin- and
hygromycin-resistance genes.
ACCESSION .
VERSION .
KEYWORDS pGreenII 0179
SOURCE http://www.biofeng.com/
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5138)
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 5138)
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..5138
/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 T-DNA repeat"
/note="left border repeat from nopaline C58 T-DNA
(truncated)"
/note="color: #ffe4c4"
polyA_signal 663..839
/note="CaMV poly(A) signal"
/note="cauliflower mosaic virus polyadenylation signal"
/note="color: #a6acb3"
CDS complement(1049..2074)
/codon_start=1
/product="hygromycin B phosphotransferase"
/note="HygR"
/note="confers resistance to hygromycin"
/note="color: #ccffcc"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
E"
promoter complement(2091..2436)
/note="CaMV 35S promoter"
/note="strong constitutive promoter from cauliflower mosaic
virus"
/note="color: #ffffff; direction: LEFT"
CDS complement(2434..2871)
/codon_start=1
/gene="lacZ (truncated)"
/product="LacZ-alpha fragment of?beta-galactosidase"
/note="lacZ-alpha"
/note="color: #993366"
/translation="MTMITPSSELTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
PSTSRGGPVPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
DRPSQQLRSLNGEWREIVNVNVIVPLLQKCQRYSLRRPKGY"
primer_bind 2658..2674
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
/note="color: #a020f0; direction: RIGHT"
promoter 2681..2699
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
/note="color: #ffffff; direction: RIGHT"
misc_feature 2708..2815
/note="MCS"
/note="pBluescript multiple cloning site"
/note="color: #99ccff"
promoter complement(2828..2846)
/note="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
/note="color: #ffffff; direction: LEFT"
primer_bind complement(2867..2883)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
/note="color: #a020f0; direction: LEFT"
protein_bind 2891..2907
/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(2915..2946)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
/note="This reverse directional feature has 3 segments:
???1:?2915?..?2921?/?#ffffff?/?-10
???2:?2922?..?2940?/?#ffffff
???3:?2941?..?2946?/?#ffffff?/?-35"
misc_feature 3221..3245
/note="RB T-DNA repeat"
/note="right border repeat from nopaline C58 T-DNA"
/note="color: #ffe4c4"
rep_origin complement(3336..3924)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/note="color: #ffff00"
CDS complement(4095..4910)
/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 tcgatctgga ttttagtact ggattttggt tttaggaatt agaaatttta ttgatagaag
721 tattttacaa atacaaatac atactaaggg tttcttatat gctcaacaca tgagcgaaac
781 cctataagaa ccctaatttc ccttatcggg aaactactca cacattattt atggagaaaa
841 tagagagaga tagatttgta gagagagact ggtgatttca gcgtaccgaa ttaattctcc
901 cgcgaccagc cgagcgagct tagcgaactg tggacgagaa ctgtgccacc aagcgtaagg
961 ccgttctctc gcatttgcct tgctaggctc gcgcgagttg ctggctgagg cgttctcgaa
1021 atcagctctt gttcggtcgg catctactct attcctttgc cctcggacga gtgctggggc
1081 gtcggtttcc actatcggcg agtacttcta cacagccatc ggtccagacg gccgcgcttc
1141 tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt gcgtcgcatc
1201 gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac caagctctga tagagttggt
1261 caagaccaat gcggagcata tacgcccgga ggcgtggcga tcctgcaagc tccggatgcc
1321 tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc cagaagaaga
1381 tgttggcgac ctcgtattgg gaatccccga acatcgcctc gctccagtca atgaccgctg
1441 ttatgcggcc attgtccgtc aggacattgt tggagccgaa atccgcgtgc acgaggtgcc
1501 ggacttcggg gcagtcctcg gcccaaagca tcagctcatc gagagcctgc gcgacggacg
1561 cactgacggt gtcgtccatc acagtttgcc agtgatacac atggggatca gcaatcgcgc
1621 atatgaaatc acgccatgta gtgtattgac cgattccttg cggtccgaat gggccgaacc
1681 cgctcgtctg gctaagatcg gccgcagcga tcgcatccat agcctccgcg accggctgaa
1741 gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt gcacggcggg
1801 agatgcaata ggtcaggctc tcgctgaact ccccaatgtc aagcacttcc ggaatcggga
1861 gcgcggccga tgcaaagtgc cgataaacat aacgatcttt gtagaaacca tcggcgcagc
1921 tatttacccg caggacatat ccacgccctc ctacatcgaa gctgaaagca cgagattctt
1981 cgccctccga gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc agaaacttct
2041 cgacagacgt cgcggtgagt tcaggctttt tcatagggat cagcttgggc tgtcctctcc
2101 aaatgaaatg aacttcctta tatagaggaa gggtcttgcg aaggatagtg ggattgtgcg
2161 tcatccctta cgtcagtgga gatgtcacat caatccactt gctttgaaga cgtggttgga
2221 acgtcttctt tttccacgat gctcctcgtg ggtgggggtc catctttggg accactgtcg
2281 gcagaggcat cttgaatgat agcctttcct ttatcgcaat gatggcattt gtaggagcca
2341 ccttcctttt ctactgtcct ttcgatgaag tgacagatag ctgggcaatg gaatccgagg
2401 aggtttcccg aaattaccct ttgttgaaaa gtctcaatag ccctttggtc ttctgagact
2461 gtatctttga catttttgga gtaggggtac gataacatta acgtttacaa tttcgcgcca
2521 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
2581 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
2641 ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact cactataggg
2701 cgaattgggt accgggcccc ccctcgaggt cgacggtatc gataagcttg atatcgaatt
2761 cctgcagccc gggggatcca ctagttctag agcggccgcc accgcggtgg agctccagct
2821 tttgttccct ttagtgaggg ttaattccga gcttggcgta atcatggtca tagctgtttc
2881 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga aghcataaag
2941 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
3001 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
3061 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
3121 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
3181 acagaatcag gggataacgc aggaaagaac atgaaggcct tgacaggata tattggcggg
3241 taaactaagt cgctgtatgt gtttgtttga gatctcatgt gagcaaaagg ccagcaaaag
3301 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
3361 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
3421 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
3481 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
3541 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
3601 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
3661 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
3721 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
3781 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaagaagagt tggtagctct
3841 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
3901 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
3961 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
4021 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgtgtaa
4081 cattggtcta gtgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
4141 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
4201 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
4261 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
4321 ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact
4381 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
4441 ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta
4501 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
4561 cctgaatcag gatattcttc taatacctgg aatgctgttt tccctgggat cgcagtggtg
4621 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
4681 tccgtcagcc agtttagtct gaccatctca tctgtaacaa cattggcaac gctacctttg
4741 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcggta gattgtcgca
4801 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
4861 gaatttaatc gcggccttga gcaagacgtt tcccgttgaa tatggctcat aacacccctt
4921 gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt
4981 gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt
5041 gctgagttga aggatcagat cacgcatctt cccgacaacg cagaccgttc cgtggcaaag
5101 caaaagttca aaatcaccaa ctggtccacc tacaacaa
//