pET-21d

PVT0062    2ug
 

pET-21d Plasmid information
Promoter: T7/lac
Replicator: ColE1 ori, F1 ori
Terminator: T7 Terminator
Plasmid classification: Escherichia coli vector; PET series expression plasmid
Plasmid size: 5440bp
Plasmid label: N-T7, C-His
Prokaryotic resistance: ampicillin Amp
Cloned strain: DH5 alpha
Culture conditions: 37 centigrade, aerobic, LB
Expression host: BL21 (DE3)
Culture conditions: 37 centigrade, aerobic, LB
Inducement: IPTG or lactose and its analogues
5'sequencing primers: T7:TAATACGACTCACTATAGGG
3'sequencing primers: T7-ter:TGCTAGTTATTGCTCAGCGG
Use:pET Plasmid
 

pET-21d plasmid Description
The pET-21a-d(+) vectors carry an N-terminal T7•Tag sequence plus an optional C-terminal His•Tag sequence. These vectors differ from pET-24a-d(+) only by their selectable marker (ampicillin vs. kanamycin resistance). 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 circular 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 T7 terminator primer.

pET-21d plasmid  Sequence

LOCUS       Exported                5440 bp ds-DNA     circular SYN 31-7-2015
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5440)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported 2015-7-31
FEATURES             Location/Qualifiers
     source          1..5440
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          244..266
                     /organism="Enterobacteria phage T7"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:10760"
     terminator      26..73
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             complement(205..237)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /note="T7 tag (gene 10 leader)"
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     RBS             244..266
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    281..305
                     /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(306..324)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        633..710
                     /gene="lacI"
                     /note="lacI promoter"
     CDS             711..1793
                     /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             2602..2793
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    2895..3037
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3223..3811)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(3982..4842)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4843..4947)
                     /gene="bla"
                     /note="AmpR promoter"
     rep_origin      complement(4974..5429)
                     /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 tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tccgacccat ttgctgtcca ccagtcatgc tagccatggt
      241 atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta tccgctcaca
      301 attcccctat agtgagtcgt attaatttcg cgggatcgag atctcgatcc tctacgccgg
      361 acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga
      421 catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt
      481 gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc
      541 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca
      601 ggagtcgcat aagggagagc gtcgagatcc cggacaccat cgaatggcgc aaaacctttc
      661 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
      721 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
      781 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
      841 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
      901 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
      961 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
     1021 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
     1081 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
     1141 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
     1201 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
     1261 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
     1321 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
     1381 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
     1441 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
     1501 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
     1561 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
     1621 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
     1681 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     1741 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     1801 gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat gcccttgaga
     1861 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     1921 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt
     1981 ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc
     2041 ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc
     2101 gagaagcagg ccattatcgc cggcatggcg gccccacggg tgcgcatgat cgtgctcctg
     2161 tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg
     2221 atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca
     2281 tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc
     2341 accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca
     2401 tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc
     2461 cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta
     2521 acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat
     2581 cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc
     2641 tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa
     2701 caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc
     2761 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     2821 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     2881 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
     2941 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatatgc ggtgtgaaat
     3001 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
     3061 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3121 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3181 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3241 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3301 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3361 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3421 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3481 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3541 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3601 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3661 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3721 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3781 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3841 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3901 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3961 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4021 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4081 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4141 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4201 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4261 gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc
     4321 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4381 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4441 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4501 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4561 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4621 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4681 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4741 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4801 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4861 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4921 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaaattgt
     4981 aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     5041 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     5101 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     5161 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     5221 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     5281 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     5341 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     5401 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
//

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Search name

pET-21d,Plasmid pET-21d,pET-21d vector