pHT304

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  • Model: PVT5009
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pHT304

PVT5009      2ug
 

pHT304 information

Promoter: Lac
Replicon: ColE1 ori
Plasmid classification: wide host series, Bacillus subtilis vector
Plasmid size: 6530bp
Plasmid Tags: LacZa
Prokaryotic resistance: Amp
Selection marker: ERM
Clone strain: DH5 alpha
Culture conditions: 37 LB, aerobic
Host: Bacillus subtilis
Induction method: IPTG induction
Primers for 5'sequencing: M13R:CAGGAAACAGCTATGACC
Primers for 3'sequencing: M13F:TGTAAAACGACGGCCAGT
Use: Bacillus subtilis
pHT304-plasmid

pHT304 description
The promoter of pHT304 vector is Lac, which can be used to induce the expression of recombinant protein in Escherichia coli DE3 engineering bacteria (DE3 BL21 or DE3 Rosetta). The tag has LacZa, which can be purified by nickel column.

 


pHT304 sequence

LOCUS       Exported File           6530 bp ds-DNA    circular SYN 29-9-2015

DEFINITION  .

ACCESSION   .

VERSION     .

KEYWORDS    Untitled

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 6530)

  AUTHORS   .

  TITLE     Direct Submission

  JOURNAL   Exported 2015-9-29

FEATURES             Location/Qualifiers

     source          1..6530

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     CDS             3862..4722

                     /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      4893..5481

                     /direction=RIGHT

                     /note="ori"

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 

                     replication"

     protein_bind    5769..5790

                     /bound_moiety="E. coli catabolite activator protein"

                     /note="CAP binding site"

                     /note="CAP binding activates transcription in the presence 

                     of cAMP."

     promoter        5805..5835

                     /note="lac promoter"

                     /note="promoter for the E. coli lac operon"

     protein_bind    5843..5859

                     /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)."

     primer_bind     5867..5883

                     /note="M13 rev"

                     /note="common sequencing primer, one of multiple similar 

                     variants"

     CDS             5879..6202

                     /codon_start=1

                     /gene="lacZ?fragment"

                     /product="LacZ-alpha fragment of beta-galactosidase"

                     /note="lacZ-alpha"

                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR

                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS

                     DAA"

     misc_feature    5896..5952

                     /gene="

                     "

                     /note="MCS"

                     /note="pUC18/19 multiple cloning site"

     primer_bind     complement(5953..5969)

                     /note="M13 fwd"

                     /note="common sequencing primer, one of multiple similar 

                     variants"

ORIGIN

        1 ccatcctcca aagttggaga gtgagtttta tgtcgcaaat attaatgttt ctggtgaacc

       61 ttatcaaatt ttcgttgatt taatagaaac atagcggtaa aattagcagt aacttaatag

      121 aacggaaatg aaaaaagcca ctctcatatg ctattggcta ccaaccttta gcgagaatga

      181 cttaatcctg tacagccata caggacttcg acttataaga ggcgccaacc tcaaataagt

      241 tatttgcctt gttttcgcga acaaggctta ttagatacac ctattgtacc gttactctac

      301 gaatatttca actagtaatt actagcattg tcatatacat aataaaacgg atataaaagg

      361 gcgttttcta tacctagaag tcttgtaaat gtacagggcg tttagatata gagaacgccc

      421 tttttgtgtt ccgttccagt ggaggctacc actttaaaaa gatggtctag tgtagccaat

      481 gcaggagagt acactcggat atcagttgtc gttgcattca actgtctgac gtaagcgagg

      541 taaaggacac aagccttgca taaaacaagc ctacgggatg taaatcctaa taatgatgat

      601 aaccaagacg ttagcggcaa aaagtgttgg gggttcaaaa taagacatga ttgtgcgact

      661 ggagttaaac agttactcgt aagcggcgat catgacactg attcacggct attcttgtac

      721 aaactttatt acaaggatat gcgggttata tagcgaatca cccgaaaggg aacggtgttg

      781 ggcgtgagaa acgcaccgta cggcgcaata caatgccaat aagctatata cggacggtat

      841 agtagttttg taagctataa ccgtttgtcg tcaatgcaac caatctcaat tcgagacctc

      901 ggcatctaag ccagtacgaa tgagtgggcg ttttaacctc gtaaattttc aacaggggtt

      961 actatgccca aaactacatt cagatttcct aacaaactcg ccagtatgaa aaccttaaga

     1021 ccttaaagtc aagggatttg aaggatttta acctcgatta gcaaaaaatg tagagtactg

     1081 aagcaactac cattaactaa gatagtgggg gattgaggaa gaatccagag ctgtttaaat

     1141 caagtgaaag acaagatgaa attaaaagaa tagtgaaaga taggggagtg gttctctatg

     1201 agaaaggaaa tggctagaga acaaaggcag cggtttattg atctattgtt agactttatg

     1261 gtaaagaatc ctcatttatt tgttaatggt acagaggatg aaagtaataa tgttgttaca

     1321 aaatgtaata gtgatattaa agaggttgcg gagtcatatt taactctttt atagtgagag

     1381 ggttaaaact aattaatatg tattaaggcc caatgttgga attattgtat ttcactaggc

     1441 aacctactta ctaaaagtaa gattatccat tagtggatgt tataatattg ggttttttaa

     1501 cacaataatc atcgcctttc ggtgtcgttt gatagaaaag taaccattag cgatgaaaaa

     1561 gtcaatataa aaagccatcc gtaaaaaacg gatggcttac cgtacatagg atcgttggta

     1621 gggcggcgta tcctacatct ctggtaactg acctagccaa tcaaatgctt gagaacggcg

     1681 gttagataag cgcgtgggga acctttccca cctcaaagat cctatatcat tattatgtta

     1741 ctttctacag gtagtatacc atgttcttat attttagtaa actccccgtt agcttaacag

     1801 gtctttgtaa gcaattaaac gtccactatt caatcgtctt tggattttcg caggaccgtt

     1861 ttttagatcg aacatagttg ataagaacaa ataaccgctt gggtccaact ttatagcaat

     1921 tagtatatgg tcatttaaaa tctttaccaa ttcaacgcta ttaggttctt taggattttg

     1981 cccgacatag tcggggtgtt caacgatatc ttttatgtgc gatgaatatt tttcataaat

     2041 accaggatgt tgtttcttta cgtgctttat aaatccggga aacattttta catcgttaga

     2101 agtgcaagtc aagttatatg tatctataat gatttgtgga agttttgcca caacagttgg

     2161 tttatttaca atcttttttt tattagccgt caaatttctc cctcatctcg tctctttata

     2221 tctttatttt atcataaagg agtatttgaa ccgtcgcgcg ggacaggttt atgataggga

     2281 tattttattg aataattgat ggtataaggg actttcatgc ttggaaagtg gggattatga

     2341 attagatgct tgtccacaat atgttccaat gtaattaaaa tttatgttcc caccttgacc

     2401 aaacatcacg tccatactta aatcgtccct cctttaatag gtaaaatatt aatttacctt

     2461 aataaaaaaa taatggataa tagtattcgt ctgaatttat ataatcaggg ggaactattg

     2521 atgctgggga tactatttac agcggcgcca tctactgatg tcgtaaagga tttgcaagat

     2581 aaagttatat cattgcagga tcatgaggta gcgtttttga acaccacgat atctaatatg

     2641 ttgatcccct tagaagcaaa cttaagagtg tgttgatagt gcagtatctt aaaattttgt

     2701 gtataatagg aattgaagtt aaattagatg ctaaaaattt gtaattaaga aggagggatt

     2761 cgtcatgttg gtattccaaa tgcgtaatgt agataaaaca tctactgttt tgaaacagac

     2821 taaaaacagt gattacgcag ataaataaat acgttagatt aattcctacc agtgactaat

     2881 cttatgactt tttaaacaga taactaaaat tacaaacaaa tcgtttaact tctgtattta

     2941 tttacagatg taatcacttc aggagtaatt acatgaacaa aaatataaaa tattctcaaa

     3001 actttttaac gagtgaaaaa gtactcaacc aaataataaa acaattgaat ttaaaagaaa

     3061 ccgataccgt ttacgaaatt ggaacaggta aagggcattt aacgacgaaa ctggctaaaa

     3121 taagtaaaca ggtaacgtct attgaattag acagtcatct attcaactta tcgacagaaa

     3181 aattaaaact gaacattcgt gtcactttaa ttcaccaaga tattctacag tttcaattcc

     3241 ctaacaaaca gaggtataaa attgttggga gtattcctta ccatttaagc acacaaatta

     3301 ttaaaaaagt ggtttttgaa agccatgcgt ctgacatcta tctgattgtt gaagaaggat

     3361 tctacaagcg taccttggat attcaccgaa cactagggtt gctcttgcac actcaagtct

     3421 cgattcagca attgcttaag ctgccagcgg aatgctttca tcctaaacca aaagtaaaca

     3481 gtgtcttaat aaaacttacc cgccatacca cagatgttcc agataaatat tggaagctat

     3541 atacgtactt tgtttcaaaa tgggtcaatc gagaatatcg tcaactgttt actaaaaatc

     3601 agtttcatca agcaatgaaa cacgccaaag taaacaattt aagtaccatt acttatgagc

     3661 aagtattgtc tatttttaat agttatctat tatttaacgg gaggaaataa ttctatgagt

     3721 cgctttttta aatttggaaa gttacacgtt actaaaggga atggagataa attattagat

     3781 atactactga cagcttccaa gaaggtaaag aggtccctag cgcctacggg gaatttgtat

     3841 cgggattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt

     3901 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat

     3961 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag

     4021 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg

     4081 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata

     4141 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat

     4201 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc

     4261 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg

     4321 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac

     4381 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact

     4441 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa

     4501 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct

     4561 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc

     4621 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga

     4681 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac

     4741 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag

     4801 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg

     4861 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc

     4921 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag

     4981 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc

     5041 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac

     5101 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc

     5161 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt

     5221 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt

     5281 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc

     5341 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt

     5401 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca

     5461 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt

     5521 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt

     5581 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag

     5641 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg

     5701 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc

     5761 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt

     5821 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat

     5881 gaccatgatt acgccaagct tgcatgcctg caggtcgact ctagaggatc cccgggtacc

     5941 gagctcgaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac

     6001 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc

     6061 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg

     6121 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac

     6181 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc

     6241 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg

     6301 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct

     6361 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg

     6421 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc

     6481 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat

//

Caution:
Product is for research use only!
 

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