质粒序列
LOCUS Exported 4320 bp ds-DNA circular SYN 28-十一月-2013
DEFINITION Mammalian expression vector for tagging proteins with a C-terminal
FLAG epitope. For other reading frames, use pCMV-Tag 4A or pCMV-Tag
4C.
ACCESSION .
VERSION .
KEYWORDS pCMV-Tag 4B
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4320)
AUTHORS Agilent Technologies
TITLE Direct Submission
JOURNAL Exported 2016年9月18日 from SnapGene Viewer 2.8.1
http://www.miaolingbio.com
COMMENT The cloned gene must include a Kozak sequence and start codon.
FEATURES Location/Qualifiers
source 1..4320
/organism="synthetic DNA construct"
/lab_host="Mammalian Cells"
/mol_type="other DNA"
enhancer 66..370
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 371..574
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 620..638
/note="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
misc_feature 651..744
/note="MCS"
/note="multiple cloning site"
CDS 745..768
/codon_start=1
/product="FLAG epitope tag, followed by an enterokinase
cleavage site"
/note="FLAG"
/translation="DYKDDDDK"
misc_feature 784..794
/note="stop codons"
/note="stop codons in all three reading frames"
promoter complement(822..840)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 1114..1235
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(1242..1697)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1724..1826
/gene="bla"
/note="AmpR promoter"
promoter 1830..2187
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
CDS 2222..3016
/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)"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
misc_feature 3248..3295
/note="HSV TK poly(A) signal"
/note="herpesvirus thymidine kinase polyadenylation signal"
rep_origin 3624..4212
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
601 gcgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg gagctccacc
661 gcggtggcgg ccgctctagc ccgggcggat cccccgggct gcaggaattc gatatcaagc
721 ttatcgatac cgtcgacact cgaggattac aaggatgacg acgataagta gggcccggta
781 ccttaattaa ttaaggtacc aggtaagtgt acccaattcg ccctatagtg agtcgtatta
841 caattcactc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggagatccaa
901 tttttaagtg tataatgtgt taaactactg attctaattg tttgtgtatt ttagattcac
961 agtcccaagg ctcatttcag gcccctcagt cctcacagtc tgttcatgat cataatcagc
1021 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
1081 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
1141 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
1201 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
1261 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
1321 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
1381 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
1441 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
1501 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
1561 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
1621 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
1681 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
1741 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
1801 gcttcaataa tattgaaaaa ggaagaatcc tgaggcggaa agaaccagct gtggaatgtg
1861 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
1921 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
1981 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
2041 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
2101 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
2161 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
2221 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
2281 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
2341 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
2401 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
2461 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
2521 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
2581 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
2641 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
2701 agaacatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
2761 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
2821 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
2881 catagcgttg gctacccgtg atattgctga agaacttggc ggcgaatggg ctgaccgctt
2941 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
3001 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
3061 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
3121 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
3181 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
3241 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
3301 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
3361 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
3421 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
3481 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
3541 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
3601 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
3661 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
3721 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
3781 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
3841 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
3901 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
3961 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
4021 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
4081 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
4141 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
4201 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
4261 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
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