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载体序列
LOCUS Exported 3889 bp ds-DNA circular SYN 13-AUG-2016
DEFINITION synthetic circular DNA
KEYWORDS pENTR-R4-pLac-Spec-R3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3889)
AUTHORS Biofeng Lab.
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..3889
/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 complement(603..727)
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR4"
/note="recombination site for the Gateway(R) LR reaction"
promoter 755..785
/note="lac promoter"
/note="promoter for the E. coli lac operon"
promoter 755..785
/note="lac UV5 promoter"
/note="E. coli lac promoter with an ""up"" mutation"
protein_bind 793..809
/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 1051..1842
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/note="SmR"
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
protein_bind 1952..2075
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR3"
/note="recombination site for the Gateway(R) LR reaction"
promoter complement(2131..2149)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2154..2170)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS 2283..3092
/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 3239..3827
/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 tcgggcccct acaggtcact aataccatct aagtagttga
601 ttcatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
661 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcaact tttctataca
721 aagttggtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
781 tattgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gcatcgctca
841 cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc
901 atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa
961 gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag
1021 ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc cgaagtatcg
1081 actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc
1141 gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg
1201 ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt
1261 ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt
1321 gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga
1381 gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat
1441 ctggctatct tgctgacaaa agcaagggaa catagcgttg ccttggtagg tccagcggcg
1501 gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta
1561 acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg
1621 tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac
1681 tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct
1741 tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc
1801 cactacgtga aaggcgagat caccaaggta gtcggcaaat aacactataa gggcgaattc
1861 tgcagatatc catcacactg gcggccgcca gtgtgatgga tatctgcaga attcgccctt
1921 catcactgca gacggggctt ttttatgcag gcaactttgt ataataaagt tgaacgagaa
1981 acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa acagactaca
2041 taatactgta aaacacaaca tatccagtca ctatgaatca actacttaga tggtattagt
2101 gacctgtact gcagggcggc cgcgatatcc cctatagtga gtcgtattac atggtcatag
2161 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
2221 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
2281 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
2341 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
2401 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
2461 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
2521 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
2581 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
2641 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
2701 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
2761 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
2821 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
2881 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
2941 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
3001 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
3061 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
3121 ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg
3181 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
3241 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
3301 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
3361 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
3421 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
3481 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
3541 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
3601 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
3661 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
3721 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
3781 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
3841 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt
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