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pENTR-L5-pLac-Spec-L2载体质粒基本信息
pENTR-L5-pLac-Spec-L2质粒图谱载体图谱和pENTR-L5-pLac-Spec-L2载体序列质粒序列多克隆位点信息
pENTR-L5-pLac-Spec-L2质粒载体简介
pENTR-L5-pLac-Spec-L2质粒序列载体序列
LOCUS Exported 3713 bp ds-DNA circular SYN 13-AUG-2016
DEFINITION synthetic circular DNA
KEYWORDS pENTR-L5-pLac-Spec-L2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3713)
AUTHORS Biofeng Lab.
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..3713
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 268..295
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 387..473
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 537..553
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 537..552
/gene="mutant version of attL"
/bound_moiety="LR Clonase(TM)"
/note="attL5"
/note="recombination site for the Gateway(R) LR reaction"
misc_feature 554..568
/note="MCS"
promoter 570..767
/note="lac"
promoter 694..724
/note="lac UV5 promoter"
/note="E. coli lac promoter with an ""up"" mutation"
promoter 694..724
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 732..748
/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)."
CDS 990..1781
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/note="SmR"
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTARLDETTRRALIDDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
protein_bind complement(1834..1933)
/gene="mutant version of attL"
/bound_moiety="LR Clonase(TM)"
/note="attL2"
/note="recombination site for the Gateway(R) LR reaction"
promoter complement(1955..1973)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1978..1994)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS 2107..2916
/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"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 3063..3651
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtatacaaa
661 agttggttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
721 attgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg catcgctcac
781 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
841 tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag
901 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg
961 gcagtcgccc taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc gaagtatcga
1021 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg
1081 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc
1141 tggttacggt gaccgcaagg cttgatgaaa caacgcggcg agctttgatc gacgaccttt
1201 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg
1261 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag
1321 aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc
1381 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg
1441 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa
1501 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt
1561 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact
1621 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt
1681 atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc
1741 actacgtgaa aggcgagatc accaaggtag tcggcaaata acactataag ggcggatgat
1801 gcagacgggg cttttttatg caggacccag ctttcttgta caaagttggc attataagaa
1861 agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta
1921 tttgccatcc agctgcaggg cggccgcgat atcccctata gtgagtcgta ttacatggtc
1981 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
2041 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
2101 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
2161 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
2221 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
2281 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
2341 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
2401 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
2461 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
2521 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
2581 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
2641 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
2701 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
2761 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
2821 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
2881 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
2941 cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt
3001 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
3061 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
3121 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
3181 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
3241 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
3301 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
3361 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
3421 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
3481 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
3541 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
3601 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
3661 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtt
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