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
