LOCUS       pTriEx-6.        5070 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Insect, bacterial, and mammalian vector, encoding a N-terminal 
            Strep-Tag(R) II-HRV 3C cassette and a cleavable C-terminal 10xHis 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    pTriEx-6
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5070)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5070)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     181..1008
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     enhancer        1076..1379
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1381..1580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     exon            1597..1698
                     /note="Exon 1"
     intron          1699..1862
     promoter        1931..1949
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1950..1974
                     /label=lac operator
                     /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        1990..2099
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     RBS             2101..2106
                     /note="ribosome binding site"
     CDS             2114..2116
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             2123..2146
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     CDS             2153..2176
                     /label=HRV 3C site
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases"
     misc_feature    2174..2218
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             2222..2239
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             2246..2272
                     /label=9xHis
                     /note="9xHis affinity tag"
     polyA_signal    2430..2485
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     terminator      2573..2620
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     2633..3338
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     protein_bind    complement(3347..3380)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(3548..4135)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4151..5008)
                     /label=AmpR
                     /note="beta-lactamase"
ORIGIN
        1 tcctgcatct tttaatcaaa tcccaagatg tgtataaacg cgccggtatg tacaggaaga
       61 ggtttatact aaactgttac attgcaaacg tggtttcgtg tgccaagtgt gaaaaccgat
      121 gtttaatcaa ggctctgacg catttctaca accacgactc caagtgtgtg ggtgaagtca
      181 tgcatctttt aatcaaatcc caagatgtgt ataaaccacc aaactgccaa aaaatgaaaa
      241 ctgtcgacaa gctctgtccg tttgctggca actgcaaggg tctcaatcct atttgtaatt
      301 attgaataat aaaacaatta taaatgtcaa atttgttttt tattaacgat acaaaccaaa
      361 cgcaacaaga acatttgtag tattatctat aattgaaaac gcgtagttat aatcgctgag
      421 gtaatattta aaatcatttt caaatgattc acagttaatt tgcgacaata taattttatt
      481 ttcacataaa ctagacgcct tgtcgtcttc ttcttcgtat tccttctctt tttcattttt
      541 ctcttcataa aaattaacat agttattatc gtatccatat atgtatctat cgtatagagt
      601 aaattttttg ttgtcataaa tatatatgtc ttttttaatg gggtgtatag taccgctgcg
      661 catagttttt ctgtaattta caacagtgct attttctggt agttcttcgg agtgtgttgc
      721 tttaattatt aaatttatat aatcaatgaa tttgggatcg tcggttttgt acaatatgtt
      781 gccggcatag tacgcagctt cttctagttc aattacacca ttttttagca gcaccggatt
      841 aacataactt tccaaaatgt tgtacgaacc gttaaacaaa aacagttcac ctcccttttc
      901 tatactattg tctgcgagca gttgtttgtt gttaaaaata acagccattg taatgagacg
      961 cacaaactaa tatcacaaac tggaaatgtc tatcaatata tagttgctga tggccggcct
     1021 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     1081 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     1141 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     1201 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagtc
     1261 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     1321 ccttacggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc
     1381 tgatgcggtt ttggcagtac accaatgggc gtggatagcg gtttgactca cggggatttc
     1441 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     1501 ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt
     1561 gggaggtcta tataagcaga cgtcgtttag tgaaccgtca gatcactaga tgctttattg
     1621 cggtagttta tcacagttaa attgctaacg ccagtctcga acttaacgtg cagaagttgg
     1681 tcgtgaggca ctgggcaggt aagtatcggg ccctttgtgc ggggggagcg gctcggggct
     1741 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     1801 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
     1861 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattggatc
     1921 ggaccgaaat taatacgact cactataggg gaattgtgag cggataacaa ttccccggag
     1981 ttaatccggg acctttaatt caacccaaca caatatatta tagttaaata agaattatta
     2041 tcaaatcatt tgtatattaa ttaaaatact atactgtaaa ttacatttta tttacaatca
     2101 aaggagatat accatggcaa gctggagcca cccgcagttc gaaaagggtg cacttgaagt
     2161 cctctttcag ggacccgggt accaggatcc tgtacaagtc gacgcggccg cagagctcgc
     2221 tctggtgcca cgcggtagtt ccgctcatca ccaccatcat caccatcacc accactaagt
     2281 gattaacctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg
     2341 gctcacaaat accactgaga tcgatctttt tccctctgcc aaaaattatg gggacatcat
     2401 gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt
     2461 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa
     2521 acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgt aactagcata
     2581 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaagcat gcggaggaaa
     2641 ttctccttga agtttccctg gtgttcaaag taaaggagtt tgcaccagac gcacctctgt
     2701 tcactggtcc ggcgtattaa aacacgatac attgttatta gtacatttat taagcgctag
     2761 attctgtgcg ttgttgattt acagacaatt gttgtacgta ttttaataat tcattaaatt
     2821 tataatcttt agggtggtat gttagagcga aaatcaaatg attttcagcg tctttatatc
     2881 tgaatttaaa tattaaatcc tcaatagatt tgtaaaatag gtttcgatta gtttcaaaca
     2941 agggttgttt ttccgaaccg atggctggac tatctaatgg attttcgctc aacgccacaa
     3001 aacttgccaa atcttgtagc agcaatctag ctttgtcgat attcgtttgt gttttgtttt
     3061 gtaataaagg ttcgacgtcg ttcaaaatat tatgcgcttt tgtatttctt tcatcactgt
     3121 cgttagtgta caattgactc gacgtaaaca cgttaaatag agcttggaca tatttaacat
     3181 cgggcgtgtt agctttatta ggccgattat cgtcgtcgtc ccaaccctcg tcgttagaag
     3241 ttgcttccga agacgatttt gccatagcca cacgacgcct attaattgtg tcggctaaca
     3301 cgtccgcgat caaatttgta gttgagcttt ttggaattgc gatcgcataa cttcgtatag
     3361 catacattat acgaagttat aagctcggaa cgctgcgctc ggtcgttcgg ctgcggcgag
     3421 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3481 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3541 tggcgttttt ccataggctc cgccccctga cgagcatcac aaaaatcgac gctcaagtca
     3601 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3661 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3721 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3781 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3841 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3901 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3961 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4021 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4081 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4141 tcctttgtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4201 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4261 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4321 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4381 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4441 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4501 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4561 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4621 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4681 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4741 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4801 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4861 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4921 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4981 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5041 atcagggtta ttgtctcatg tccgcgcgtt
//
