pTWIN2

价格:900元

联系方式:I47-825O-882O

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载体基本信息

出品公司: NEB
载体名称: pTWIN2
质粒类型: 大肠杆菌表达载体
表达水平:
启动子: T7
克隆方法: 多克隆位点,限制性内切酶
载体大小: 7192 bp
5' 测序引物及序列: T7:TAATACGACTCACTATAGGG
3' 测序引物及序列: --
载体标签: --
载体抗性: Ampicillin
筛选标记: --
备注: --
产品目录号: E6901
稳定性: --
组成型: --
病毒/非病毒: 非病毒

载体质粒图谱和多克隆位点信息

pTWIN2载体图谱
pTWIN2多克隆位点

pTWIN2载体特征

载体简介

pTWIN2 is an E. coli expression vector which can be used with the IMPACT? Kit (NEB #E6901) (1). A polylinker in the vector is designed for the in-frame fusion of a target gene between the modified Ssp DnaB (2) and Mth RIR1 inteins (3). The presence of the chitin binding domain from Bacillus circulans (4,5) facilitates purification. pTWIN vectors are designed for protein purification or for the isolation of proteins with an N-terminal cysteine and/or a C-terminal thioester (1). The double-stranded vector is 7192 base pairs in length.

载体序列

LOCUS       pTWIN2                  7192 bp    DNA     circular     09-NOV-2011
DEFINITION  Cloning vector pTWIN2, complete sequence.
ACCESSION
VERSION
KEYWORDS    .
SOURCE      Cloning vector pTWIN2
  ORGANISM  Cloning vector pTWIN2
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 7192)
  AUTHORS   Evans,T.C. Jr., Benner,J. and Xu,M.-Q.
  TITLE     The in vitro ligation of bacterially expressed protein using an
            intein from Methanobacterium thermoautotrophicum
  JOURNAL   J Biol Chem 274, 3923-3926 (1999)
   PUBMED   9933578
REFERENCE   2  (bases 1 to 7192)
  AUTHORS   Mathys,S., Evans,T.C. Jr., Chute,I.C., Wu,H., Chong,S., Benner,J.,
            Liu,X.-Q. and Xu,M.-Q.
  TITLE     Characterization of a self-splicing mini-intein and its conversion
            into autocatalytic N- and C-terminal cleavage elements: facile
            production of protein building blocks for protein ligation
  JOURNAL   Gene 231, 1-13 (1999)
   PUBMED   10231563
REFERENCE   3  (bases 1 to 7192)
  AUTHORS   Evans,T.C. Jr., Benner,J. and Xu,M.-Q.
  TITLE     The cyclization and polymerization of bacterially-expressed
            proteins using modified self-splicing inteins
  JOURNAL   J Biol Chem 274, 18359-18363 (1999)
   PUBMED   10373440
REFERENCE   4  (bases 1 to 7192)
  AUTHORS   Perler,F.B.
  TITLE     InBase, the intein database
  JOURNAL   Nucleic Acids Res 30 (1), 383-384 (2002)
   PUBMED   11752343
REFERENCE   5  (bases 1 to 7192)
  AUTHORS   Ghosh,I. and Xu,M.Q.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-OCT-2007) Research Department, New England Biolabs,
            240 County Road, Ipswich, MA 01938, USA
FEATURES             Location/Qualifiers
     source          1..7192
                     /organism="Cloning vector pTWIN2"
                     /mol_type="other DNA"
     gene            140..1000
                     /gene="bla"
     CDS             140..1000
                     /gene="bla"
                     /note="ampR (confers resistance to ampicillin)"
                     /codon_start=1
                     /product="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     sig_peptide     140..208
                     /gene="bla"
                     /note="required for secretion to the periplasm; cleaved
                     off to form the mature beta-lactamase protein"
     rep_origin      1042..1555
                     /note="M13 origin of replication (+ -)"
     rep_origin      1666..2254
                     /note="pMB1 origin of replication (clockwise) (RNAII -35
                     to RNA/DNA switch point)"
     gene            complement(2623..2814)
                     /gene="rop"
     CDS             complement(2623..2814)
                     /gene="rop"
                     /exception="alternative start codon"
                     /codon_start=1
                     /transl_table=11
                     /product="ROP protein"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH
                     ADELYRSCLARFGDDGENL"
     gene            complement(3371..4453)
                     /gene="lacI"
     CDS             complement(3371..4453)
                     /gene="lacI"
                     /exception="alternative start codon"
                     /codon_start=1
                     /transl_table=11
                     /product="lac repressor (LacI)"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE
                     LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERS
                     GVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
                     FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREG
                     DWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT
                     EDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNT
                     QTASPRALADSLMQLARQVSRLESGQ"
     primer_bind     5637..5656
                     /note="T7 universal primer sequence (NEB #S1248S)"
     promoter        5637..5654
                     /note="T7 promoter (transcript start 5654 clockwise)"
     protein_bind    5656..5680
                     /note="lac operator"
                     /bound_moiety="LacI"
     RBS             5710..5716
                     /note="RBS for expression CDS"
     gene            5725..7044
                     /gene="expression CDS"
     CDS             5725..7044
                     /gene="expression CDS"
                     /note="chitin binding domain (CBD) sequence 5725..5901;
                     Ssp DNAB intein sequence 5941..6402; multiple cloning site
                     (SapI-SapI) 6403..6444; Mth RIR1 intein sequence
                     6445..6846; CBD sequence 6886..7044"
                     /codon_start=1
                     /product="CBD/intein/intein/CBD fusion"
                     /translation="MKIEEGKLTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSL
                     AGWEPSNVPALWQLQNNGNNGLELRESGAISGDSLISLASTGKRVSIKDLLDEKDFEI
                     WAINEQTMKLESAKVSRVFCTGKKLVYILKTRLGRTIKATANHRFLTIDGWKRLDELS
                     LKEHIALPRKLESSSLQLSPEIEKLSQSDIYWDSIVSITETGVEEVFDLTVPGPHNFV
                     ANDIIVHNGRAMGGREFLEGSSCVSGDTIVMTSGGPRTVAELEGKPFTALIRGSGYPC
                     PSGFFRTCERDVYDLRTREGHCLRLTHDHRVLVMDGGLEWRAAGELERGDRLVMDDAA
                     GEFPALATFRGLRGAGRQDVYDATVYGASAFTANGFIVHACGEQPGTGLNSGLTTNPG
                     VSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNVPALWQLQ"
     primer_bind     6296..6315
                     /note="Ssp DnaB intein forward primer sequence (NEB
                     #S1269S)"
     misc_feature    6403..6444
                     /gene="expression CDS"
                     /note="multiple cloning site (SapI-SapI)"
     primer_bind     complement(6505..6522)
                     /note="Mth RIR1 intein reverse primer sequence (NEB
                     #S1270S)"
     terminator      7064..7186
                     /note="T7 Tphi transcription terminator"
BASE COUNT     1785 a   1803 c   1934 g   1670 t
ORIGIN
        1 aactacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
       61 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
      121 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
      181 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
      241 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
      301 ccttgagagt tttcgccccg aagaacgttc tccaatgatg agcactttta aagttctgct
      361 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
      421 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
      481 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
      541 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
      601 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
      661 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
      721 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
      781 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
      841 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
      901 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
      961 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     1021 atatatactt tagattgatt taccccggtt gataatcaga aaagccccaa aaacaggaag
     1081 attgtataag caaatattta aattgtaaac gttaatattt tgttaaaatt cgcgttaaat
     1141 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     1201 tcaaaagaat agcccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     1261 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     1321 ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     1381 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     1441 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     1501 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtaaaag
     1561 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     1621 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1681 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1741 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     1801 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     1861 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     1921 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     1981 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2041 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     2101 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2161 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2221 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2281 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2341 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2401 cgagcgcagc gagtcagtga gcgaggaagc tatggtgcac tctcagtaca atctgctctg
     2461 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     2521 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     2581 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     2641 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gcagcgattc
     2701 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     2761 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     2821 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     2881 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     2941 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcc
     3001 gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt
     3061 ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat
     3121 tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc
     3181 gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt
     3241 cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     3301 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     3361 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3421 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     3481 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     3541 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     3601 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac
     3661 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     3721 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     3781 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     3841 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     3901 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     3961 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     4021 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     4081 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     4141 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     4201 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     4261 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     4321 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     4381 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     4441 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     4501 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
     4561 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
     4621 aatggtgcat gccggcatgc cgccctttcg tcttcaagaa ttaattccca attccccagg
     4681 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     4741 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag
     4801 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggaa ttaattcccc
     4861 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     4921 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
     4981 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gaattaattc
     5041 cccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     5101 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     5161 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggaattaa
     5221 ttccccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     5281 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     5341 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggaat
     5401 taattcccca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     5461 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     5521 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     5581 aattggggat cggaattaat tcccggttta aaccggggat ctcgatcccg cgaaattaat
     5641 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt
     5701 taactttaag aaggagatat acatatgaaa atcgaagaag gtaaactgac aaatcctggt
     5761 gtatccgctt ggcaggtcaa cacagcttat actgcgggac aattggtcac atataacggc
     5821 aagacgtata aatgtttgca gccccacacc tccttggcag gatgggaacc atccaacgtt
     5881 cctgccttgt ggcagcttca aaacaacggt aacaacggtc tcgaactgcg cgagtccgga
     5941 gctatctctg gcgatagtct gatcagcctg gctagcacag gaaaaagagt ttctattaaa
     6001 gatttgttag atgaaaaaga ttttgaaata tgggcaatta atgaacagac gatgaagcta
     6061 gaatcagcta aagttagtcg tgtattttgt actggcaaaa agctagttta tattctaaaa
     6121 actcgactag gtagaactat caaggcaaca gcaaatcata gatttttaac tattgatggt
     6181 tggaaaagat tagatgagct atctttaaaa gagcatattg ctctaccccg taaactagaa
     6241 agctcctctt tacaattgtc accagaaata gaaaagttgt ctcagagtga tatttactgg
     6301 gactccatcg tttctattac ggagactgga gtcgaagagg tttttgattt gactgtgcca
     6361 ggaccacata actttgtcgc gaatgacatc attgtacaca acggaagagc catgggcggc
     6421 cgcgaattcc tcgagggctc ttcctgcgta tccggtgaca ccattgtaat gactagtggc
     6481 ggtccgcgca ctgtggctga actggagggc aaaccgttca ccgcactgat tcgcggctct
     6541 ggctacccat gcccctcagg tttcttccgc acctgtgaac gtgacgtata tgatctgcgt
     6601 acacgtgagg gtcattgctt acgtttgacc catgatcacc gtgttctggt gatggatggt
     6661 ggcctggaat ggcgtgccgc gggtgaactg gaacgcggcg accgcctggt gatggatgat
     6721 gcagctggcg agtttccggc actggcaacc ttccgtggcc tgcgtggcgc tggccgccag
     6781 gatgtttatg acgctactgt ttacggtgct agcgcattca ctgctaatgg cttcattgta
     6841 cacgcatgcg gcgagcagcc cgggaccggt ctgaactcag gcctcacgac aaatcctggt
     6901 gtatccgctt ggcaggtcaa cacagcttat actgcgggac aattggtcac atataacggc
     6961 aagacgtata aatgtttgca gccccacacc tccttggcag gatgggaacc atccaacgtt
     7021 cctgccttgt ggcagcttca atgactgcag gaaggggatc cggctgctaa caaagcccga
     7081 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     7141 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg at
//

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