LOCUS       Exported                7413 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 7413)
  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..7413
                     /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(2207..2209)
                     /codon_start=1
                     /product="epitope tag from a T7 major capsid protein"
                     /label=T7 tag
                     /translation="M"
     RBS             2218..2223
                     /note="ribosome binding site"
     protein_bind    2254..2278
                     /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(2279..2297)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2606..2683
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2684..3766
                     /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             4575..4766
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     rep_origin      complement(5196..5784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5955..6815)
                     /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(6816..6920)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6947..7402)
                     /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 ggatccccag cttgcagagc agctctcgta gccatttcaa
     1741 aaacctctct cactccatct ttggtctttg ctgaacactc catgtaccca aaagcgccaa
     1801 tcctgtttgc catatctctg ccttcttcag gtttcaccgg ctcctgcttc atcttggcta
     1861 gctcccgcct tgtgtgctca tcattccgaa gatccttctt attcccaacc aggatgatgg
     1921 gcacgttggg acagaaatgc ttgacttctg gggtccactt ttctgggatg ttttctaaac
     1981 tatcagggct gtcgatggaa aaacacatca gtataacatc ggtatctggg taggagaggg
     2041 gcctcaggcg atcataatct tccagcccag ctgtgtccca caaagccaac tctacctgct
     2101 ttccatccac ctcgatatct gccacatagt tctcaaacac tgtgggcaca tacacctctg
     2161 ggaactgatc cttgctgaag actatgagca agcatgtctt tccacacata tgtatatctc
     2221 cttcttaaag ttaaacaaaa ttatttctag aggggaattg ttatccgctc acaattcccc
     2281 tatagtgagt cgtattaatt tcgcgggatc gagatctcga tcctctacgc cggacgcatc
     2341 gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc
     2401 gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg
     2461 gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt
     2521 gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg
     2581 cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct ttcgcggtat
     2641 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
     2701 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
     2761 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
     2821 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
     2881 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
     2941 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
     3001 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
     3061 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
     3121 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
     3181 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
     3241 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
     3301 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
     3361 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
     3421 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
     3481 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt
     3541 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
     3601 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
     3661 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     3721 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     3781 atgtaagtta gctcactcat taggcaccgg gatctcgacc gatgcccttg agagccttca
     3841 acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg
     3901 tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg
     3961 aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct
     4021 tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc
     4081 aggccattat cgccggcatg gcggccccac gggtgcgcat gatcgtgctc ctgtcgttga
     4141 ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg
     4201 agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg
     4261 tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta
     4321 tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat
     4381 taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg
     4441 ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta
     4501 tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt
     4561 acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca
     4621 gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag
     4681 acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt
     4741 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     4801 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     4861 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta
     4921 tgcggcatca gagcagattg tactgagagt gcaccatata tgcggtgtga aataccgcac
     4981 agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg
     5041 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5101 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5161 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5221 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5281 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5341 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5401 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5461 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5521 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5581 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     5641 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5701 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5761 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5821 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5881 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5941 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6001 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6061 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6121 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6181 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6241 aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt
     6301 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6361 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6421 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6481 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6541 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6601 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6661 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6721 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6781 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6841 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6901 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaaat tgtaaacgtt
     6961 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     7021 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     7081 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     7141 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     7201 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     7261 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     7321 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     7381 aatgcgccgc tacagggcgc gtcccattcg cca
//
