LOCUS       pTriEx-3_Hygro.        6732 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Insect, bacterial, and mammalian vector with a CMV promoter and a 
            hygromycin resistance marker, for expressing proteins with a 
            C-terminal HSV-8xHis cassette.
ACCESSION   .
VERSION     .
KEYWORDS    pTriEx-3 Hygro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6732)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6732)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6732
                     /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"
     misc_feature    2112..2219
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             2114..2116
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             2225..2257
                     /label=HSV tag
                     /note="HSV (herpes simplex virus) epitope tag"
     CDS             2264..2287
                     /label=8xHis
                     /note="8xHis affinity tag"
     misc_feature    2341..2804
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2843..3859
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     polyA_signal    4092..4147
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     terminator      4235..4282
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     4295..5000
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     protein_bind    complement(5009..5042)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(5210..5797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5813..6670)
                     /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 accatggcga tatcccggga gctcgtggat ccgaattctc agatctcggc
     2161 gcgcctgcag gtcgacggta ccggttcgaa gcttgcggcc gcacagctgt atacacgtgc
     2221 aagccagcca gaactcgccc cggaagaccc cgaggatctc gagcaccacc atcaccatca
     2281 ccatcactaa gtgattaacc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg
     2341 gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct
     2401 cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc
     2461 ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga
     2521 caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc
     2581 ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctca cctcaagcgt
     2641 attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggaa tctgatctgg
     2701 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     2761 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataagcttgc cacaacccgt
     2821 accaaagatg gatagatccg gaaagcctga actcaccgcg acgtctgttg agaagtttct
     2881 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg
     2941 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
     3001 tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc
     3061 ggaagtgctt gacattgggg aatttagcga gagcctgacc tattgcatct cccgccgtgc
     3121 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcaaccggt
     3181 cgcggaggct atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc
     3241 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
     3301 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc
     3361 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
     3421 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat
     3481 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
     3541 gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga
     3601 gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta
     3661 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc
     3721 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc
     3781 cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac
     3841 tcgtccgagg gcaaaggaat agagtagatg ccgaccgaac aagagctgat ttcgagaacg
     3901 cctcagccag caactcgcgc gagcctagca aggcaaatgc gagagaacgg ccttacgctt
     3961 ggtggcacag ttctcgtcca cagttcgcta accaatgccc tggctcacaa ataccactga
     4021 gatcgatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct
     4081 gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     4141 tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt
     4201 ggtttagagt ttggcaacat atgcccatat gtaactagca taaccccttg gggcctctaa
     4261 acgggtcttg aggggttttt tgctgaaagc atgcggagga aattctcctt gaagtttccc
     4321 tggtgttcaa agtaaaggag tttgcaccag acgcacctct gttcactggt ccggcgtatt
     4381 aaaacacgat acattgttat tagtacattt attaagcgct agattctgtg cgttgttgat
     4441 ttacagacaa ttgttgtacg tattttaata attcattaaa tttataatct ttagggtggt
     4501 atgttagagc gaaaatcaaa tgattttcag cgtctttata tctgaattta aatattaaat
     4561 cctcaataga tttgtaaaat aggtttcgat tagtttcaaa caagggttgt ttttccgaac
     4621 cgatggctgg actatctaat ggattttcgc tcaacgccac aaaacttgcc aaatcttgta
     4681 gcagcaatct agctttgtcg atattcgttt gtgttttgtt ttgtaataaa ggttcgacgt
     4741 cgttcaaaat attatgcgct tttgtatttc tttcatcact gtcgttagtg tacaattgac
     4801 tcgacgtaaa cacgttaaat agagcttgga catatttaac atcgggcgtg ttagctttat
     4861 taggccgatt atcgtcgtcg tcccaaccct cgtcgttaga agttgcttcc gaagacgatt
     4921 ttgccatagc cacacgacgc ctattaattg tgtcggctaa cacgtccgcg atcaaatttg
     4981 tagttgagct ttttggaatt gcgatcgcat aacttcgtat agcatacatt atacgaagtt
     5041 ataagctcgg aacgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5101 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5161 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5221 tccgccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5281 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5341 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5401 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5461 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5521 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5581 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5641 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5701 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5761 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttgt taccaatgct
     5821 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5881 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5941 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6001 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6061 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6121 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6181 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6241 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6301 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6361 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6421 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6481 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6541 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6601 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6661 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6721 tgtccgcgcg tt
//
