别名: |
FABP11; H-FABP; M-FABP; MDGI; O-FABP |
质粒分类: |
基因文库质粒 |
原核抗性: |
卡那霉素Kan |
克隆菌株: |
大肠杆菌DH5α |
培养条件: |
37℃ |
pET28a-H-FABP是一个人源基因大肠表达质粒。
LOCUS Exported 5749 bp ds-DNA circular SYN 27-JAN-2018
FEATURES Location/Qualifiers
source 1..5749
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 26..73
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(140..157)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
misc_feature complement(179..577)
/label=H-FABP
CDS complement(587..619)
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/label=T7 tag (gene 10 leader)
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
CDS complement(623..640)
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS complement(650..667)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
RBS 686..708
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 723..747
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(748..766)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 1075..1152
/gene="lacI"
/label=lacI promoter
CDS 1153..2235
/codon_start=1
/gene="lacI"
/product="lac repressor"
/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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
CDS 3044..3235
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 3337..3479
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(3665..4253)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 4375..5190
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin complement(5283..5738)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgg
181 cctctttctc ataggtgcgg gtacacacgg cggtgccgtg tgtcagtgtc aggatcagct
241 tgccgtcgat cagctcgcgc actaaggtgg tctcctggcc gtcccatttt tgcaggtgca
301 ccagcttacc gccgtccagg gtcacgatgc tcttcacctt gcggtcgtcg gcggtggtct
361 cgtcgaactc cacgcccagc ttgaagctga tctcggtgtt cttgaaggtg ctgtgggtct
421 tcagggtcag aatgtcgccg tttttctcga tgatggtggt cggcttggtc atactggcca
481 cctggcgtgt ggcaaaaccc acgcccaggc tcttcatgta gtcgtcaaag ttcttggagt
541 ccactaactt ccaggtgccc aggaaagcgt ccaccatgga tccgcgaccc atttgctgtc
601 caccagtcat gctagccata tggctgccgc gcggcaccag gccgctgctg tgatgatgat
661 gatgatggct gctgcccatg gtatatctcc ttcttaaagt taaacaaaat tatttctaga
721 ggggaattgt tatccgctca caattcccct atagtgagtc gtattaattt cgcgggatcg
781 agatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
841 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
901 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
961 gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac
1021 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat cccggacacc
1081 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
1141 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
1201 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
1261 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
1321 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
1381 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
1441 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
1501 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
1561 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
1621 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
1681 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
1741 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
1801 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
1861 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
1921 attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc
1981 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
2041 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
2101 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
2161 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
2221 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcaccggg
2281 atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
2341 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
2401 gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat
2461 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg
2521 tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccccacg
2581 ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac
2641 tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac
2701 gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga
2761 aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg
2821 gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat
2881 ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa
2941 ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat
3001 cattaccccc atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa
3061 aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact
3121 caacgagctg gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga
3181 tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
3241 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
3301 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
3361 cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg
3421 caccatatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
3481 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
3541 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
3601 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
3661 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
3721 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
3781 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
3841 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
3901 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
3961 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
4021 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
4081 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
4141 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
4201 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
4261 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
4321 tggtcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
4381 catattcaac gggaaacgtc ttgctctagg ccgcgattaa attccaacat ggatgctgat
4441 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga
4501 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
4561 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
4621 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
4681 gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
4741 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
4801 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
4861 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
4921 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
4981 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
5041 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
5101 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
5161 tttcatttga tgctcgatga gtttttctaa gaattaattc atgagcggat acatatttga
5221 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
5281 tgaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
5341 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
5401 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
5461 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
5521 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
5581 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
5641 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
5701 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgcca