LOCUS       pTriEx-4.        5238 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Insect, bacterial, and mammalian vector with a CMV promoter, 
            encoding an N-terminal 6xHis-S-Tag and a C-terminal HSV-8xHis.
ACCESSION   .
VERSION     .
KEYWORDS    pTriEx-4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5238)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5238)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5238
                     /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             2120..2137
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             2147..2191
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             2207..2224
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             2243..2257
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             2381..2413
                     /label=HSV tag
                     /note="HSV (herpes simplex virus) epitope tag"
     CDS             2420..2443
                     /label=8xHis
                     /note="8xHis affinity tag"
     polyA_signal    2598..2653
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     terminator      2741..2788
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     2801..3506
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     protein_bind    complement(3515..3548)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(3716..4303)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4319..5176)
                     /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 accatggcac accatcacca ccatcactct tctggtaaag aaaccgctgc
     2161 tgcgaaattt gaacgccagc acatggactc gccaccgcct tctggtctgg tcccccgggg
     2221 cagcgcaggt tctggtacga ttgatgacga cgacaagagt ccgggcttct cctcaacgat
     2281 atctgagctc gtggatccga attctcagat ctcggcgcgc ctgcaggtcg acggtaccgg
     2341 ttcgaagctt gcggccgcac agctgtatac acgtgcaagc cagccagaac tcgccccgga
     2401 agaccccgag gatctcgagc accaccatca ccatcaccat cactaagtga ttaacctcag
     2461 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac
     2521 cactgagatc gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     2581 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     2641 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     2701 gtatttggtt tagagtttgg caacatatgc ccatatgtaa ctagcataac cccttggggc
     2761 ctctaaacgg gtcttgaggg gttttttgct gaaagcatgc ggaggaaatt ctccttgaag
     2821 tttccctggt gttcaaagta aaggagtttg caccagacgc acctctgttc actggtccgg
     2881 cgtattaaaa cacgatacat tgttattagt acatttatta agcgctagat tctgtgcgtt
     2941 gttgatttac agacaattgt tgtacgtatt ttaataattc attaaattta taatctttag
     3001 ggtggtatgt tagagcgaaa atcaaatgat tttcagcgtc tttatatctg aatttaaata
     3061 ttaaatcctc aatagatttg taaaataggt ttcgattagt ttcaaacaag ggttgttttt
     3121 ccgaaccgat ggctggacta tctaatggat tttcgctcaa cgccacaaaa cttgccaaat
     3181 cttgtagcag caatctagct ttgtcgatat tcgtttgtgt tttgttttgt aataaaggtt
     3241 cgacgtcgtt caaaatatta tgcgcttttg tatttctttc atcactgtcg ttagtgtaca
     3301 attgactcga cgtaaacacg ttaaatagag cttggacata tttaacatcg ggcgtgttag
     3361 ctttattagg ccgattatcg tcgtcgtccc aaccctcgtc gttagaagtt gcttccgaag
     3421 acgattttgc catagccaca cgacgcctat taattgtgtc ggctaacacg tccgcgatca
     3481 aatttgtagt tgagcttttt ggaattgcga tcgcataact tcgtatagca tacattatac
     3541 gaagttataa gctcggaacg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3601 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3661 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3721 ataggctccg ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3781 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3841 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3901 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3961 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4021 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4081 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4141 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4201 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4261 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgttacc
     4321 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4381 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4441 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4501 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4561 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4621 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4681 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4741 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4801 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4861 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4921 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4981 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5041 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5101 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5161 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5221 gtctcatgtc cgcgcgtt
//
