LOCUS       pTriEx-3_Neo.        6445 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Insect, bacterial, and mammalian vector with a CMV promoter and a 
            neomycin resistance marker, for expressing proteins with a 
            C-terminal HSV-8xHis cassette.
ACCESSION   .
VERSION     .
KEYWORDS    pTriEx-3 Neo
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6445)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6445)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6445
                     /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    2386..2848
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2884..3684
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     polyA_signal    3805..3860
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     terminator      3948..3995
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     4008..4713
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     protein_bind    complement(4722..4755)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(4923..5510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5526..6383)
                     /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 gtaattccgg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     2401 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     2461 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     2521 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     2581 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     2641 gagttggata gttgtggaaa gagtcaaatg gctcacctca agcgtattca acaaggggct
     2701 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     2761 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     2821 ggttttcctt tgaaaaacac gatgataagc ttgccacaac ccgcgataat tcctgcagcc
     2881 aatatgggat cggccattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     2941 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3001 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3061 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3121 tgcgcagcag tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3181 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3241 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3301 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3361 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3421 cgtatgcccg acggcgatga tctcgtcgtg actcatggcg atgcctgctt gccgaatatc
     3481 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3541 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     3601 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     3661 tatcgccttc ttgacgagtt cttctgaggg gatcaattct ctagccaatg ccctggctca
     3721 caaataccac tgagatcgat ctttttccct ctgccaaaaa ttatggggac atcatgaagc
     3781 cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt
     3841 ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc
     3901 agaatgagta tttggtttag agtttggcaa catatgccca tatgtaacta gcataacccc
     3961 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa agcatgcgga ggaaattctc
     4021 cttgaagttt ccctggtgtt caaagtaaag gagtttgcac cagacgcacc tctgttcact
     4081 ggtccggcgt attaaaacac gatacattgt tattagtaca tttattaagc gctagattct
     4141 gtgcgttgtt gatttacaga caattgttgt acgtatttta ataattcatt aaatttataa
     4201 tctttagggt ggtatgttag agcgaaaatc aaatgatttt cagcgtcttt atatctgaat
     4261 ttaaatatta aatcctcaat agatttgtaa aataggtttc gattagtttc aaacaagggt
     4321 tgtttttccg aaccgatggc tggactatct aatggatttt cgctcaacgc cacaaaactt
     4381 gccaaatctt gtagcagcaa tctagctttg tcgatattcg tttgtgtttt gttttgtaat
     4441 aaaggttcga cgtcgttcaa aatattatgc gcttttgtat ttctttcatc actgtcgtta
     4501 gtgtacaatt gactcgacgt aaacacgtta aatagagctt ggacatattt aacatcgggc
     4561 gtgttagctt tattaggccg attatcgtcg tcgtcccaac cctcgtcgtt agaagttgct
     4621 tccgaagacg attttgccat agccacacga cgcctattaa ttgtgtcggc taacacgtcc
     4681 gcgatcaaat ttgtagttga gctttttgga attgcgatcg cataacttcg tatagcatac
     4741 attatacgaa gttataagct cggaacgctg cgctcggtcg ttcggctgcg gcgagcggta
     4801 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4861 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4921 tttttccata ggctccgccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4981 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5041 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5101 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5161 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5221 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5281 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5341 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5401 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5461 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5521 tgttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5581 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5641 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5701 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5761 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5821 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5881 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5941 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6001 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6061 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6121 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6181 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6241 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6301 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6361 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6421 ggttattgtc tcatgtccgc gcgtt
//
