• 规格：20μl质粒
• 启动子：T7/lac
• 复制子：ColE1 ori，F1 ori
• 终止子：T7 terminator
• 质粒分类：大肠杆菌载体；PET系列表达质粒
• 质粒大小：7309bp
• 质粒标签：C-HSV, N-6×His, C-6×His, N-Thrombin, N-Nus, N-EK
• 原核抗性：氨苄青霉素Amp
• 克隆菌株：DH5α
• 表达宿主：BL21(DE3)
• 诱导方式：IPTG或乳糖及其类似物
• 5测序引物：T7:TAATACGACTCACTATAGGG
• 3测序引物：T7-ter:TGCTAGTTATTGCTCAGCGG
• 质粒简介：        The pET-44 vectors are designed for cloning and high-level expression of peptide sequences fused with the 495 aa Nus•Tag™ protein. Compared to the pET-43.1 series, the pET-44 vectors encode an additional N-terminal His•Tag. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circle map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single stranded DNA that corresponds to the coding strand. Therefore, single stranded sequencing should be performed using the COLIDOWN primer . Vector encoded sequence can be completely removed when cloning into the PshA I or Sma I sites (as shown below) by cleaving the Nus•Tag fusion protein with enterokinase or thrombin, respectively.
  
• 质粒图谱1： png/3-1610121P32aZ.png

质粒序列： 点击查看序列

LOCUS       Exported                7309 bp ds-DNA     circular SYN 06-SEP-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pET-44c
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7309)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, September 6, 2016 from SnapGene Viewer 3.1.4
            http://www.fenghbio.cn
FEATURES             Location/Qualifiers
     source          1..7309
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      26..73
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     terminator      260..346
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(512..529)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             complement(536..568)
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /note="HSV tag"
                     /translation="QPELAPEDPED"
     CDS             complement(679..693)
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /note="enterokinase site"
                     /translation="DDDDK"
     CDS             complement(712..729)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /note="thrombin site"
                     /translation="LVPRGS"
     CDS             complement(745..789)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
                     /note="S-Tag"
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(799..816)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             complement(832..2316)
                     /codon_start=1
                     /gene="nusA"
                     /product="transcription elongation protein (N utilization
                     substance protein A)"
                     /note="NusA"
                     /note="highly soluble in E. coli"
                     /translation="MNKEILAVVEAVSNEKALPREKIFEALESALATATKKKYEQEIDV
                     RVQIDRKSGDFDTFRRWLVVDEVTQPTKEITLEAARYEDESLNLGDYVEDQIESVTFDR
                     ITTQTAKQVIVQKVREAERAMVVDQFREHEGEIITGVVKKVNRDNISLDLGNNAEAVIL
                     REDMLPRENFRPGDRVRGVLYSVRPEARGAQLFVTRSKPEMLIELFRIEVPEIGEEVIE
                     IKAAARDPGSRAKIAVKTNDKRIDPVGACVGMRGARVQAVSTELGGERIDIVLWDDNPA
                     QFVINAMAPADVASIVVDEDKHTMDIAVEAGNLAQAIGRNGQNVRLASQLSGWELNVMT
                     VDDLQAKHQAEAHAAIDTFTKYLDIDEDFATVLVEEGFSTLEELAYVPMKELLEIEGLD
                     EPTVEALRERAKNALATIAQAQEESLGDNKPADDLLNLEGVDRDLAFKLAARGVCTLED
                     LAEQGIDDLADIEGLTDEKAGALIMAARNICWFGDEA"
     CDS             complement(2323..2340)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     protein_bind    2397..2421
                     /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(2422..2440)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2753..2830
                     /gene="lacI"
                     /note="lacI promoter"
     CDS             2831..3913
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /note="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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     CDS             4485..4676
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     rep_origin      complement(5106..5694)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of
                     replication"
     promoter        5775..5867
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             5868..6728
                     /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"
     rep_origin      complement(6843..7298)
                     /direction=LEFT
                     /note="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 tcagcggata catatttgaa tgtatttaga aaaataaaca
      121 aaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttaatttgat
      181 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt
      241 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
      301 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc
      361 cctactctcg catggggaga ccccacacta ccatcggcgc tacggcgttt cacttctgag
      421 ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc
      481 agcctaggaa cgcccaactt aattaacatt agtggtggtg gtggtggtgc tcgagatcct
      541 ctgggtcttc aggagcgagt tctggctggc ttgcacgtgt atacagctgt gcggccgcaa
      601 gcttcgaacg cgtatcgatg gtaccgtcga cgtcctgcag gcgcgcctgt acagaattcg
      661 gatccacgag ctcggactct tgtcgtcgtc atcaatcgta ccagaacccg cgctgccccg
      721 ggggaccaga ccagttggcg gtggcgagtc catgtgctgg cgttcaaatt tcgcagcagc
      781 ggtttcttta cccgcggagt gatggtgatg gtgatgacca gaaccactag tcgcttcgtc
      841 accgaaccag caaatattac gggcagccat aatcagtgct ccggcttttt cgtcggtcaa
      901 cccttcgata tcagccagat catcaatgcc ctgttcggcg agatcttcca gcgtacaaac
      961 gccacgggcg gccagtttga atgccaaatc acgatctacc ccttcaaggt tcagcagatc
     1021 gtcagccggt ttgttatcac cgaggctttc ttcctgggcc tgtgcaatgg tggccagtgc
     1081 atttttagca cgctcgcgca gtgcttcaac ggtcggctca tcaaggcctt cgatttccaa
     1141 cagctctttc atcggcacat aggccaattc ttccagcgtc gagaagcctt cttctaccag
     1201 aacagtcgcg aagtcttcgt cgatgtcgag atatttggtg aaggtgtcga tcgctgcgtg
     1261 cgcttccgcc tgatgcttag cttgcaggtc gtcaacggtc atcacgttga gttcccaacc
     1321 gctcagttgc gaagccagac gcacgttctg accgttacgg ccaatcgcct gcgccagatt
     1381 accggcttca acggcgatgt ccatggtgtg tttatcttca tccaccacga tagaagcaac
     1441 gtctgccggt gccattgcgt taatcacgaa ctgcgccggg ttatcatccc acaggacgat
     1501 atcgatacgc tcgccaccca gttcagtaga caccgcctga acacgcgcgc cacgcatacc
     1561 tacgcaagca cctaccggat cgatacgttt atcgttggtt ttcaccgcga ttttcgcacg
     1621 agaacccgga tcgcgagccg ctgctttaat ttcaatcact tcttcgccga tttctggcac
     1681 ttcaatacgg aacagttcga tcagcatttc cggcttggaa cgagtgacga acagttgcgc
     1741 gccacgcgct tccgggcgaa cggaatagag cacgccacga acgcggtcgc cagggcggaa
     1801 gttttcacgc ggcagcatat cttcgcgcag gatcacggct tcagcgttgt tgcccagatc
     1861 cagagagatg ttgtcgcggt ttactttttt caccacgccg gtgatgattt caccttcgtg
     1921 ttcacggaac tgatcaacca ccatcgcacg ttcggcttca cgcactttct gcacgataac
     1981 ctgttttgcc gtctgggtag tgatacggtc aaaggtaaca gactcaatct gatcttcaac
     2041 gtaatcgccc aggttcaggc tttcatcttc ataacgtgcg gcttcaaggg tgatttcctt
     2101 ggtcggctgg gtgacttcat caacaactaa ccagcgacgg aaagtgtcaa aatcaccgct
     2161 tttgcgatcg atctgtacgc ggacgtcgat ctcttgttca tattttttct ttgttgctgt
     2221 cgccagcgcg ctttccaatg cttcgaaaat cttctcgcga ggtagcgcct tttcattgga
     2281 tacggcttca actacagcca aaatttcttt gttcatacta gaatggtgat ggtggtgatg
     2341 gctgctgccc atatgtatat ctccttctta aagttaaaca aaattatttc tagaggggaa
     2401 ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg atcgagatcg
     2461 atctcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
     2521 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
     2581 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc
     2641 gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta
     2701 ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat
     2761 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
     2821 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
     2881 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
     2941 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
     3001 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
     3061 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
     3121 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
     3181 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
     3241 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
     3301 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
     3361 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
     3421 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
     3481 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
     3541 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
     3601 taccgagtcc gggctgcgcg ttggtgcgga catctcggta gtgggatacg acgataccga
     3661 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
     3721 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
     3781 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
     3841 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     3901 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcaccgggat
     3961 ctcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca
     4021 tgactagcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta
     4081 ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa
     4141 cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg
     4201 aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct
     4261 ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga
     4321 tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta
     4381 accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta
     4441 tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa
     4501 aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac
     4561 tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg
     4621 atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     4681 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     4741 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
     4801 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt
     4861 gcaccatata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     4921 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4981 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5041 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5101 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5161 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5221 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5281 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5341 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5401 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5461 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5521 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     5581 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5641 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5701 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5761 ttggtcatga cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5821 acaataaccc tgataaatgc ttcaatcatg attgaaaaag gaagagtatg agtattcaac
     5881 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5941 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     6001 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     6061 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     6121 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     6181 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     6241 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     6301 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     6361 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     6421 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     6481 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     6541 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     6601 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6661 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6721 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     6781 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgcca tcgtgccgcc
     6841 agaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc