LOCUS       Exported                6223 bp ds-DNA     circular SYN 21-MAR-2019
DEFINITION  Bacterial vector for expression of N-terminally 6xHis-tagged 
            proteins with a thrombin site. For other reading frames, use 
            pET-28b(+) or pET-28c(+).
ACCESSION   .
VERSION     .
KEYWORDS    pET-28a(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6223)
  AUTHORS   MilliporeSigma (Novagen)
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 20, 2021 from SnapGene Viewer 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6223
                     /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"
                     /note="color: #ffffff"
     primer_bind     139..187
                     /label=hGTF2B-F-XhoI
                     /note="color: black; sequence: 
                     AGTGGTGGTGGTGGTGGTGCTCGAGttatagctgtggtagtttgtccac; added: 
                     2019-03-22"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     misc_feature    158..163
                     /label=MCS
                     /note="multiple cloning site"
                     /note="color: #99ccff"
     CDS             complement(164..1114)
                     /codon_start=1
                     /label=hTF2B
                     /note="color: #993366"
                     /translation="MASTSRLDALPRVTCPNHPDAILVEDYRAGDMICPECGLVVGDRV
                     IDVGSEWRTFSNDKATKDPSRVGDSQNPLLSDGDLSTMIGKGTGAASFDEFGNSKYQNR
                     RTMSSSDRAMMNAFKEITTMADRINLPRNIVDRTNNLFKQVYEQKSLKGRANDAIASAC
                     LYIACRQEGVPRTFKEICAVSRISKKEIGRCFKLILKALETSVDLITTGDFMSRFCSNL
                     CLPKQVQMAATHIARKAVELDLVPGRSPISVAAAAIYMASQASAEKRTQKEIGDIAGVA
                     DVTIRQSYRLIYPRAPDLFPTDFKFDTPVDKLPQL"
     primer_bind     complement(1099..1174)
                     /label=hGTF2B-R-NcoI
                     /note="color: black; sequence: 
                     ACTTTAAGAAGGAGATATACCATGGGCAGCAGCCATCATCATCATCATCACAGCAGCGG
                     Catggcgtctaccagcc; added: 2019-03-22"
     CDS             complement(1124..1141)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     CDS             complement(1151..1153)
                     /codon_start=1
                     /product="start codon"
                     /label=ATG
                     /note="color: #993366"
                     /translation="M"
     RBS             1161..1166
                     /note="ribosome binding site"
                     /note="color: #a6acb3"
     protein_bind    1197..1221
                     /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)."
                     /note="color: #31849b"
     promoter        complement(1222..1240)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     promoter        1549..1626
                     /gene="lacI"
                     /label=lacI promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             1627..2709
                     /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)."
                     /note="color: #993366"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     CDS             3518..3709
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /label=rop
                     /note="color: #993366"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     rep_origin      complement(4139..4727)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             4849..5664
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /note="color: #ccffcc"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      complement(5757..6212)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagttatagc tgtggtagtt
      181 tgtccactgg ggtgtcaaat ttgaagtctg taggaaacag atctggggct cgaggataga
      241 tcagtctata ggactgtctg attgtaacat cagcaacacc agcaatatct ccaatttctt
      301 tttgggtcct cttttcagct gatgcctgtg aggccatgta aatagctgcc gctgccacag
      361 agatggggct cctcccagga accaagtcca attccacagc tttacgggct atatgtgtag
      421 ctgccatctg tacttgttta ggaagacaaa ggttggaaca gaacctggac atgaagtccc
      481 cagttgtaat caaatccaca ctggtttcta gcgctttcaa aataagttta aaacaccgac
      541 caatttcttt cttagaaatt cgtgatacgg cacatatttc tttaaatgtc ctaggaaccc
      601 cttcttgtct acaggcaata tagagacaag cagaagctat agcatcatta gctcttccct
      661 tcaggctctt ctgttcatat acttgcttga ataaattatt tgttcgatca actatatttc
      721 gaggtagatt gattctgtct gccatggtag tgatttcttt gaatgcattc atcattgccc
      781 gatcagaact gctcattgtt ctccgattct ggtacttaga attgccaaat tcgtcaaaac
      841 ttgcagctcc tgtgcccttg ccaatcatgg tagacaaatc tccatcactc agaagaggat
      901 tctgagaatc tccaactcga gatggatctt ttgttgcttt gtcattgctg aaagttcgcc
      961 attcagatcc cacatcaata acccggtcac ctacaaccaa gccacattca ggacagatca
     1021 tatcaccggc tctgtagtcc tccactaaaa tcgcatctgg atggtttgga catgtgactc
     1081 ttggaagagc atccaaacgg ctggtagacg ccatgccgct gctgtgatga tgatgatgat
     1141 ggctgctgcc catggtatat ctccttctta aagttaaaca aaattatttc tagaggggaa
     1201 ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg atcgagatct
     1261 cgatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg
     1321 gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga
     1381 gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca
     1441 tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg
     1501 gctgcttcct aatgcaggag tcgcataagg gagagcgtcg agatcccgga caccatcgaa
     1561 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
     1621 gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
     1681 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
     1741 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc
     1801 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa
     1861 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg
     1921 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc
     1981 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct
     2041 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt
     2101 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt
     2161 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg
     2221 gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc
     2281 gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt
     2341 cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc
     2401 gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac
     2461 agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa
     2521 accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg
     2581 ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct
     2641 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     2701 gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cgggatctcg
     2761 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
     2821 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
     2881 agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct
     2941 gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc
     3001 caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccc cacgggtgcg
     3061 catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta
     3121 gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc
     3181 gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg
     3241 gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct gctggctacc
     3301 ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag tgatttttct
     3361 ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca gtaaccgggc
     3421 atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg gtatcattac
     3481 ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg aaaaaaccgc
     3541 ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga
     3601 gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct
     3661 ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     3721 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     3781 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     3841 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     3901 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
     3961 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4021 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4081 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4141 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4201 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4261 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4321 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4381 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4441 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4501 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4561 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4621 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4681 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4741 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4801 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
     4861 caacgggaaa cgtcttgctc taggccgcga ttaaattcca acatggatgc tgatttatat
     4921 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat
     4981 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     5041 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     5101 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccgggaaa
     5161 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     5221 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     5281 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     5341 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     5401 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     5461 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     5521 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     5581 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     5641 ttgatgctcg atgagttttt ctaagaatta attcatgagc ggatacatat ttgaatgtat
     5701 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaaat
     5761 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     5821 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     5881 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     5941 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     6001 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     6061 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     6121 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     6181 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg cca
//
