LOCUS       Exported                7401 bp ds-DNA     circular SYN 25-DEC-2023
DEFINITION  Bacterial vector for inducible expression of N-terminally T7-tagged 
            proteins. For other reading frames, use pET-21a(+) or pET-21c(+).
ACCESSION   .
VERSION     .
KEYWORDS    pET-21b(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7401)
  AUTHORS   MilliporeSigma (Novagen)
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jan 2, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7401
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /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    167..1270
                     /label=MBP
     CDS             complement(1271..1291)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(1298..1618)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             complement(2195..2197)
                     /codon_start=1
                     /product="epitope tag from a T7 major capsid protein"
                     /label=T7 tag
                     /translation="M"
     RBS             2206..2211
                     /note="ribosome binding site"
     protein_bind    2242..2266
                     /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(2267..2285)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2594..2671
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2672..3754
                     /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             4563..4754
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     rep_origin      complement(5184..5772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5943..6803)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6804..6908)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6935..7390)
                     /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 gagctacgag ctggaattag
      181 tctgcgcgtc tttcagggct tcatcgacag tctgacgacc gctggcggcg ttgatcaccg
      241 cagtacgcac ggcataccag aaagcggaca tctgcgggat gttcggcatg atttcacctt
      301 tctgggcgtt ttccatagtg gcggcaatac gtggatcttt cgccaactct tcctcgtaag
      361 acttcagcgc tacggcaccc agcggtttgt ctttattaac cgcttccaga ccttcatcag
      421 tcagcagata gttttcgagg aactcttttg ccagctcttt gttcggactg gcggcgttaa
      481 tacctgcgct cagcacgcca acgaacggtt tggatggttg acccttgaag gtcggcagta
      541 ccgttacacc ataattcact ttgctggtgt cgatgttgga ccatgcccac gggccgttga
      601 tggtcatcgc tgtttcgcct ttattaaagg cagcttctgc gatggagtaa tcggtgtctg
      661 cattcatgtg tttgttttta atcaggtcaa ccaggaaggt cagacccgct ttcgcgccag
      721 cgttatccac gcccacgtct ttaatgtcgt acttgccgtt ttcatacttg aacgcataac
      781 ccccgtcagc agcaatcagc ggccaggtga agtacggttc ttgcaggttg aacatcagcg
      841 cgctcttacc tttcgctttc agttctttat ccagcgccgg gatctcttcc caggtttttg
      901 gcgggttcgg cagcaggtct ttgttataaa tcagcgataa cgcttcaaca gcgatcgggt
      961 aagcaatcag cttgccgttg taacgtacgg catcccaggt aaacggatac agcttgtcct
     1021 ggaacgcttt gtccggggtg atttcagcca acaggccaga ttgagcgtag ccaccaaagc
     1081 ggtcgtgtgc ccagaagata atgtcagggc catcgccagt tgccgcaacc tgtgggaatt
     1141 tctcttccag tttatccgga tgctcaacgg tgactttaat tccggtatct ttctcgaatt
     1201 tcttaccgac ttcagcgaga ccgttatagc ctttatcgcc gttaatccag attaccagtt
     1261 taccttcttc gccctggaag tacaggttct caagcttttc cagttttaga agctccacat
     1321 cgaagacgag agtggcatgt ggtgggatga tgcctgggtg cccagtggca ccataggcat
     1381 aatctggaga tatagtcagt ttggctctct gacccacact catctgggca accccttctt
     1441 cccagcctcg gatcacctcc tgcttgccta gcataaactt aaagggcttg tttctgtccc
     1501 gggaggaatc aaatttcttt ccatcttcaa gcatcccggt gtagtgcacc acgcaggtct
     1561 ggccgcgctt ggggaaggtg cgcccgtctc ctggggagat ggtttccacc tgcactccgc
     1621 cccgggacgc gtacgagatc tgacctccgc cagatgagcc acctccctgc aggcgcgccg
     1681 tgtacacgag ctcccaattg ggatccccac catctttgct catcttttct ttatgttttc
     1741 gaatttctcg aactaatgta tagaaggcat catcaacacc ctgtcttgtc tttgctgatg
     1801 tttcaataaa aggaattcca taacttcttg ctaagtcctg agcctgtttt gtgtctactg
     1861 ttctggaagg caaatcacat ttatttccta ctaggaccat aggtacatct tcagagtcct
     1921 taactctttt aatttgttct ctataatggt gaatatcttc aaatgattta gtattattta
     1981 tggcaaatac acaaagaaag ccctccccag tcctcatgta ctggtccctc attgcactgt
     2041 actcctccag acctgctgtg tcgagaatat ccaagagaca ggtttctcca tcaattacta
     2101 cttgcttcct gtaggaatcc tctattgttg gatcatattc gtccacaaaa tgattctgaa
     2161 ttagctgtat cgtcaaggca ctcttgccta cgcccatatg tatatctcct tcttaaagtt
     2221 aaacaaaatt atttctagag gggaattgtt atccgctcac aattccccta tagtgagtcg
     2281 tattaatttc gcgggatcga gatctcgatc ctctacgccg gacgcatcgt ggccggcatc
     2341 accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat
     2401 cgggctcgcc acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc
     2461 gtggccgggg gactgttggg cgccatctcc ttgcatgcac cattccttgc ggcggcggtg
     2521 ctcaacggcc tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag
     2581 cgtcgagatc ccggacacca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg
     2641 cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc
     2701 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
     2761 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
     2821 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
     2881 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
     2941 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
     3001 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
     3061 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
     3121 ctctgaccag acacccatca acagtattat tttctcccat gaagacggta cgcgactggg
     3181 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
     3241 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
     3301 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
     3361 gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc
     3421 gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt
     3481 agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa
     3541 acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg
     3601 ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct
     3661 ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     3721 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc
     3781 tcactcatta ggcaccggga tctcgaccga tgcccttgag agccttcaac ccagtcagct
     3841 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
     3901 tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc
     3961 gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg
     4021 ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg
     4081 ccggcatggc ggccccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag
     4141 gctggcgggg ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa
     4201 gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc
     4261 gtgtttcgta aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct
     4321 gcatcgcagg atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct
     4381 ggcattgacc ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac
     4441 cctcacaacg ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc
     4501 tctctcgttt catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat
     4561 cagtgaccaa acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat
     4621 taacgcttct ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat
     4681 cgcttcacga ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg
     4741 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     4801 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
     4861 ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga
     4921 gcagattgta ctgagagtgc accatatatg cggtgtgaaa taccgcacag atgcgtaagg
     4981 agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5041 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5101 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5161 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5221 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5281 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5341 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5401 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5461 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5521 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5581 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     5641 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5701 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5761 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5821 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5881 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5941 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6001 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6061 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6121 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6181 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6241 aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6301 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6361 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6421 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6481 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6541 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6601 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6661 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6721 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6781 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6841 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6901 gttccgcgca catttccccg aaaagtgcca cctgaaattg taaacgttaa tattttgtta
     6961 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     7021 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     7081 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     7141 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     7201 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     7261 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     7321 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     7381 cagggcgcgt cccattcgcc a
//
