质粒序列
LOCUS Exported 3466 bp ds-DNA circular SYN 09-JAN-2013
DEFINITION Vector for bacterial expression of 6xHis-tagged proteins that can be
processed by TAGZyme.
ACCESSION .
VERSION .
KEYWORDS TAGZyme pQE-1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3466)
AUTHORS Qiagen
TITLE Direct Submission
JOURNAL Exported Wednesday, September 14, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
COMMENT A blunt coding sequence can be inserted at the PvuII site, allowing
TAGZyme to remove the 6xHis tag and subsequent glutamine.
FEATURES Location/Qualifiers
source 1..3466
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
promoter 10..54
/note="T5 promoter"
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 30..46
/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)."
protein_bind 62..78
/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)."
RBS 101..106
/note="ribosome binding site"
CDS 115..117
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
CDS 121..138
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
misc_feature 139..197
/note="MCS"
/note="multiple cloning site"
misc_feature 197..207
/note="stop codons"
/note="stop codons in all three reading frames"
terminator 213..307
/note="lambda t0 terminator"
/note="transcription terminator from phage lambda"
CDS 351..1010
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
terminator 1075..1161
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
rep_origin complement(1642..2230)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2401..3261)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3262..3366)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaaa
121 catcaccatc accatcacca gctgcatgcg ggagctcatg cggccgcggg tacccacgtg
181 tcgacctgca gaagcttaat tagctgagct tggactcctg ttgatagatc cagtaatgac
241 ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg
301 tgagaatcca agctagcttg gcgagatttt caggagctaa ggaagctaaa atggagaaaa
361 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg
421 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctcgat attacggcct
481 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg
541 cccgcctgat gaatgctcat ccggaatttc gtatggcaat gaaagacggt gagctggtga
601 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat
661 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg
721 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt
781 tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg
841 acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc
901 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
961 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag
1021 gcagttattg gtgcccttaa acgcctgggg taatgactct ctagcttgag gcatcaaata
1081 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
1141 gctctcctga gtaggacaaa tccgccctct agagctgcct cgcgcgtttc ggtgatgacg
1201 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
1261 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
1321 ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga
1381 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
1441 aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
1501 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
1561 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
1621 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
1681 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
1741 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
1801 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
1861 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
1921 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
1981 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
2041 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
2101 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
2161 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
2221 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
2281 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
2341 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
2401 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
2461 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
2521 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
2581 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
2641 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
2701 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
2761 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
2821 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
2881 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
2941 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
3001 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
3061 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
3121 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
3181 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
3241 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
3301 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
3361 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
3421 attaacctat aaaaataggc gtatcacgag gccctttcgt cttcac
//