NovoPro
Cart 2

pTWIN2 vector (V012685#)

Basic Vector Information

pTWIN2 is an E. coli expression vector which can be used with the IMPACTâ„¢ Kit. A polylinker in the vector is designed for the in-frame fusion of a target gene between the modified Ssp DnaB and Mth RIR1 inteins. The presence of the chitin binding domain from Bacillus circulans 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. The double-stranded vector is 7192 base pairs in length.

Basic Vector Information
Vector Name pTWIN2 Antibiotic Resistance Ampicillin
Length 7192 bp Type Bacterial expression vector
Source NEB Copy Number High copy number
Promoter T7 Fusion Tag intein
Expression Method IPTG induced

pTWIN2 vector Vector Map

pTWIN2 vector Sequence

Copy Sequence

Download SnapGene Map(.dna)

Download GeneBank File(.gb)

LOCUS       Exported                7192 bp ds-DNA     circular SYN 09-DEC-2020
DEFINITION  Cloning vector pTWIN2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    pTWIN2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7192)
  AUTHORS   Evans TC Jr., Benner J, 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 TC Jr., Chute IC, Wu H, Chong S, Benner J, Liu X-Q.,
            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 TC Jr., Benner J, 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 FB.
  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, Xu MQ.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-OCT-2007) Research Department, New England Biolabs, 
            240 County Road, Ipswich, MA 01938, USA
REFERENCE   6  (bases 1 to 7192)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, December 9, 2020 from SnapGene 3.2.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7192
                     /organism="Cloning vector pTWIN2"
                     /mol_type="other DNA"
     CDS             140..1000
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="bla"
                     /note="ampR (confers resistance to ampicillin)"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            140..1000
                     /gene="bla"
                     /note="bla"
     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)"
     CDS             complement(2623..2814)
                     /codon_start=1
                     /transl_table=11
                     /exception="alternative start codon"
                     /gene="rop"
                     /product="ROP protein"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     gene            complement(2623..2814)
                     /gene="rop"
                     /note="rop"
     CDS             complement(3371..4453)
                     /codon_start=1
                     /transl_table=11
                     /exception="alternative start codon"
                     /gene="lacI"
                     /product="lac repressor (LacI)"
                     /note="lacI"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(3371..4453)
                     /gene="lacI"
                     /note="lacI"
     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
                     /bound_moiety="LacI"
                     /note="LacI lac operator"
                     /note="lac operator"
     RBS             5710..5716
                     /note="RBS for expression CDS"
     CDS             5725..7044
                     /codon_start=1
                     /gene="expression CDS"
                     /product="CBD/intein/intein/CBD fusion"
                     /note="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"
                     /translation="MKIEEGKLTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLA
                     GWEPSNVPALWQLQNNGNNGLELRESGAISGDSLISLASTGKRVSIKDLLDEKDFEIWA
                     INEQTMKLESAKVSRVFCTGKKLVYILKTRLGRTIKATANHRFLTIDGWKRLDELSLKE
                     HIALPRKLESSSLQLSPEIEKLSQSDIYWDSIVSITETGVEEVFDLTVPGPHNFVANDI
                     IVHNGRAMGGREFLEGSSCVSGDTIVMTSGGPRTVAELEGKPFTALIRGSGYPCPSGFF
                     RTCERDVYDLRTREGHCLRLTHDHRVLVMDGGLEWRAAGELERGDRLVMDDAAGEFPAL
                     ATFRGLRGAGRQDVYDATVYGASAFTANGFIVHACGEQPGTGLNSGLTTNPGVSAWQVN
                     TAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNVPALWQLQ"
     gene            5725..7044
                     /gene="expression CDS"
                     /note="expression CDS"
     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"

This page is informational only.