pET28a- AFP

$216.00

  • Model: PVT0595
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pET28a-AFP

 

Alternative name

pET28a-AFP, pET28a-AFP Plasmid, pET28a-AFP vector

 

pET28a-AFP Information

Alias: NM_001134.2, AFPD; FETA; HPAFP

 

Plasmid classification: gene library plasmids

 

Prokaryotic resistance: kanamycin Kan

 

Cloned strain: Escherichia coli DH5 alpha

 

Culture conditions: 37 degrees centigrade

 

 

pET28a-AFP Description

PET28a-AFP is a human source gene large intestine expression plasmid.

 

 

pET28a-AFP Sequence

LOCUS       Exported                7238 bp ds-DNA     circular SYN 08-MAY-2017

FEATURES             Location/Qualifiers

     source          1..7238

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     terminator      26..73

                     /label=T7 terminator

                     /note="transcription terminator for bacteriophage T7 RNA 

                     polymerase"

     CDS             complement(140..157)

                     /codon_start=1

                     /product="6xHis affinity tag"

                     /label=6xHis

                     /translation="HHHHHH"

     misc_feature    complement(213..2042)

                     /label=AFP

                     /note="Homo sapiens alpha fetoprotein (AFP), mRNA

                     NCBI Reference Sequence: NM_001134.2"

     CDS             complement(2076..2108)

                     /codon_start=1

                     /product="leader peptide from bacteriophage T7 gene 10"

                     /label=T7 tag (gene 10 leader)

                     /note="promotes efficient translation in E. coli"

                     /translation="MASMTGGQQMG"

     CDS             complement(2112..2129)

                     /codon_start=1

                     /product="thrombin recognition and cleavage site"

                     /label=thrombin site

                     /translation="LVPRGS"

     CDS             complement(2139..2156)

                     /codon_start=1

                     /product="6xHis affinity tag"

                     /label=6xHis

                     /translation="HHHHHH"

     RBS             2175..2197

                     /note="efficient ribosome binding site from bacteriophage 

                     T7 gene 10 (Olins and Rangwala, 1989)"

     protein_bind    2212..2236

                     /label=lac operator

                     /bound_moiety="lac repressor encoded by 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)."

     promoter        complement(2237..2255)

                     /label=T7 promoter

                     /note="promoter for bacteriophage T7 RNA polymerase"

     promoter        2564..2641

                     /gene="lacI"

                     /label=lacI promoter

     CDS             2642..3724

                     /codon_start=1

                     /gene="lacI"

                     /product="lac repressor"

                     /label=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             4533..4724

                     /codon_start=1

                     /gene="rop"

                     /product="Rop protein, which maintains plasmids at low copy

                     number"

                     /label=rop

                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA

                     DELYRSCLARFGDDGENL"

     misc_feature    4826..4968

                     /label=bom

                     /note="basis of mobility region from pBR322"

     rep_origin      complement(5154..5742)

                     /direction=LEFT

                     /label=ori

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

                     replication"

     CDS             5864..6679

                     /codon_start=1

                     /gene="aph(3')-Ia"

                     /product="aminoglycoside phosphotransferase"

                     /label=KanR

                     /note="confers resistance to kanamycin in bacteria or G418 

                     (Geneticin(R)) in eukaryotes"

                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG

                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK

                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA

                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI

                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"

     rep_origin      complement(6772..7227)

                     /direction=LEFT

                     /label=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 gagcggccgc cagtgtgatg

      181 gatatctgca gaattgccct tcccgaagta atttaaactc ccaaagcagc acgagttttt

      241 gaaatcagtt tttgtccctc ttcagcaaag cagacttcct gttcctggcc ttggcagcat

      301 ttctccaaca ggcctgagaa atctgcaatg acagcctcaa gttgttcctc tgttatttgt

      361 ggcttttgct tcacaaggtt aatgagaaac tcttgcttca tcgtttgcag cgctacaccc

      421 tgagcttggc acagatcctt atggaaaatg aacttgtcat cagagaatgc aggagggaca

      481 tatgtttcat ccaccaccaa gctgctgaag catggcctcc tgttggcata tgaagaagtg

      541 cagcactggc caacaccagg gtttactgga gtcatttcat gtctgataca taagtgtccg

      601 ataataatgt cagccgctcc ctcgccacag gccaatagtt tgtcctcact gagttggcaa

      661 caagtggctg ctgtggctgc catttttctg gtgatggcca tcagctccga cgaggtcagc

      721 tggggggctt tctttgtgta agcaacgaga aacgcatttt gtaagtaata ttctcctagt

      781 ttctggaaga ggccgcagct tcgctttgcc aatgcttggc tctcctggat gtatttctgt

      841 aattcttctt ctcctttatc ttggcattca agagggtttt cagtctggaa acacttctcc

      901 aataactcct ggtatccttt agcaactctt agaattactg agacagcaag ctgaggatgt

      961 cttcttgaat attcatgaac aaaacttgcc aagaagatat ttttttcccc tgaagaaaat

     1021 tggttaaaat ctctatctcc taaaaacctg tttagatttg gagatagacc ttcaggtttt

     1081 tcatcatttt ctgcatgaat tatacattga ccacgttcca gcgtggtcag tttgcagcat

     1141 tctgttattt tgtttgacag agtgtcttgt tgagaacata tgtaggacat gattttttcc

     1201 ccatcctgca gacaatccag cacatctcct ctgcaacagt gctcatgtac atgggccaca

     1261 tccaggacta gtttctggat ttcagtaaaa ttaactttgg taaacttctg actcagttta

     1321 gtaacagtta tggcttggaa agttcgggtc ccaaaatttt tcattactgc acatgcatgt

     1381 tgatttaaca agctgctttc tcttaattct tttgtaactg ttgctgcctt tgtttggaag

     1441 cattcaactg cattttcagc tttgcagcaa gatggaatta ttttgtcata gcgagcagcc

     1501 caaagaagaa ttgtaggtgc atacaggaag ggatgccttc ttgctatctc ataaatgaat

     1561 ttgttcatga atgtctccct gtcttcttca tatgcttcac agcttgtgac aggttctgga

     1621 acttggaaaa gtgggatcga tgctggagtg ggctttttgt gtgcaagaaa acagttatgt

     1681 cttccctctt cactttggct gcagcagtct gaatgtccgt acttctccaa aatttctttc

     1741 tcatggcaaa gttcttccag aaaggcaggt agctggtttt ctaaacaccc tgaagactgt

     1801 tcatctccag tgggtttctc aattgcagtc aatgcatctt tcaccatttt gcttacttcc

     1861 ttgtaagtgg cttcttgaac aaactgggca aaaaatatgg tagccaggtc agctaaactt

     1921 atctctgcag tacattggta agaatccaat atggaagcta ttccatattc atttctatgc

     1981 agtgttctgg attcagtaaa atttagtagg aaaattaaaa aaattgattc cacccacttc

     2041 atggttggtc gacggagctc gaattcggat ccgcgaccca tttgctgtcc accagtcatg

     2101 ctagccatat ggctgccgcg cggcaccagg ccgctgctgt gatgatgatg atgatggctg

     2161 ctgcccatgg tatatctcct tcttaaagtt aaacaaaatt atttctagag gggaattgtt

     2221 atccgctcac aattccccta tagtgagtcg tattaatttc gcgggatcga gatctcgatc

     2281 ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc

     2341 tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct

     2401 tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc

     2461 ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc

     2521 ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca tcgaatggcg

     2581 caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa

     2641 tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt

     2701 ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc

     2761 ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca

     2821 gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt

     2881 cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga

     2941 acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag

     3001 tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg

     3061 cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat

     3121 tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca

     3181 gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg

     3241 ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg

     3301 gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac

     3361 tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc

     3421 cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc

     3481 atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag

     3541 cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc

     3601 cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg

     3661 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca

     3721 gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga tctcgaccga

     3781 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg

     3841 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc

     3901 tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc

     3961 ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca

     4021 aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccccacgg gtgcgcatga

     4081 tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga

     4141 atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct

     4201 gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt

     4261 cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg

     4321 gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt

     4381 cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt

     4441 catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca

     4501 tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta

     4561 acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg

     4621 acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc

     4681 gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg

     4741 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt

     4801 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag

     4861 tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatatg

     4921 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct

     4981 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac

     5041 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga

     5101 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat

     5161 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac

     5221 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct

     5281 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg

     5341 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg

     5401 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt

     5461 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg

     5521 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac

     5581 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga

     5641 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt

     5701 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt

     5761 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaac

     5821 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg

     5881 ggaaacgtct tgctctaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta

     5941 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa

     6001 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac

     6061 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca

     6121 ttttatccgt actcctg

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