質(zhì)粒簡(jiǎn)介
pJET1.2質(zhì)粒是一個(gè)基因克隆載體。
質(zhì)粒序列
LOCUS Exported 2974 bp ds-DNA circular SYN 15-十月-2016
DEFINITION Positive selection cloning vector with a lethal insert that allows
for efficient recovery of blunt-ended PCR products.
ACCESSION EF694056
VERSION .
KEYWORDS pJET1.2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2974)
AUTHORS Thermo Scientific (Fermentas)
TITLE Direct Submission
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..2974
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
CDS complement(16..753)
/codon_start=1
/gene="eco47IR"
/product="mutant version of the Eco47I restriction
endonuclease"
/note="Eco47I/T7"
/note="cleaves double-stranded DNA at GGWCC sites in the
absence of protective methylation"
/translation="MSKETSFVKNAEELAKQKMDAINPELSSKFKFLIKFLSQFPEACS
KPRSKKMQNKVGQEEHIEYLARSFHESRLPRKPTPPTTVPDEVVSIVLNISFNIQPENL
ERIKEEHRFSMAAENIVGDLLERYLAEKLEPSGRSGGRSPYSESYYAGWIWCSGTSVKA
VDFIHYDEKNNEWNLLQVKNRDNTENSSSSKIRDNTTIKKWFRTYSQRDATNWDNFPDE
VSSKNLNEEDFRAFVKNYLAKII"
promoter 305..323
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 328..422
/note="MCS"
/note="multiple cloning site with a central Eco32I (EcoRV)
site"
protein_bind 782..798
/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)."
promoter complement(806..836)
/note="lac UV5 promoter"
/note="E. coli lac promoter with an ""up"" mutation"
rep_origin complement(1163..1751)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1922..2782)
/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(2783..2887)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc
61 ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc
121 acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact
181 cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt
241 ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc
301 ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt
361 ttcagcaaga tatctttcta gaagatctcc tacaatattc tcagctgcca tggaaaatcg
421 atgttcttct tttattctct caagattttc aggctgtata ttaaaactta tattaagaac
481 tatgctaacc acctcatcag gaaccgttgt aggtggcgtg ggttttcttg gcaatcgact
541 ctcatgaaaa ctacgagcta aatattcaat atgttcctct tgaccaactt tattctgcat
601 tttttttgaa cgaggtttag agcaagcttc aggaaactga gacaggaatt ttattaaaaa
661 tttaaatttt gaagaaagtt cagggttaat agcatccatt ttttgctttg caagttcctc
721 agcattctta acaaaagacg tctcttttga catgtttaaa gtttaaacct cctgtgtgaa
781 attgttatcc gctcacaatt ccacacatta tacgagccgg aagcataaag tgtaaagcct
841 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg ccaattgctt
901 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
961 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
1021 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
1081 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
1141 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
1201 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
1261 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
1321 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
1381 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
1441 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
1501 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
1561 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
1621 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
1681 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
1741 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
1801 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
1861 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
1921 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
1981 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
2041 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
2101 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
2161 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
2221 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
2281 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
2341 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
2401 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
2461 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
2521 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
2581 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
2641 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
2701 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
2761 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
2821 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
2881 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
2941 cattaaccta taaaaatagg cgtatcacga ggcc
//