LOCUS       pTriEx-2_Hygro.        7107 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Insect, bacterial, and mammalian vector with a hygromycin resistance
            marker, encoding an N-terminal 6xHis-S-Tag cassette and a C-terminal
            HSV-8xHis cassette.
ACCESSION   .
VERSION     .
KEYWORDS    pTriEx-2 Hygro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7107)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7107)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7107
                     /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        1140..1443
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1449..1726
                     /label=chicken beta-actin promoter
     exon            1727..1819
                     /note="Exon 1"
     intron          1820..2081
     promoter        2150..2168
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2169..2193
                     /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        2209..2318
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     RBS             2320..2325
                     /note="ribosome binding site"
     CDS             2333..2335
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             2339..2356
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             2366..2410
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             2426..2443
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             2462..2476
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             2600..2632
                     /label=HSV tag
                     /note="HSV (herpes simplex virus) epitope tag"
     CDS             2639..2662
                     /label=8xHis
                     /note="8xHis affinity tag"
     misc_feature    2716..3179
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             3218..4234
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     polyA_signal    4467..4522
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     terminator      4610..4657
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     4670..5375
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     protein_bind    complement(5384..5417)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(5585..6172)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6188..7045)
                     /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 tggccggccg
     1021 taatgagacg cacaaactaa tatcacaaac tggaaatgtc tatcaatata tagttgctct
     1081 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     1141 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     1201 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     1261 tgggtggact atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     1321 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     1381 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     1441 atgcatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc
     1501 acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg
     1561 gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg
     1621 gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag
     1681 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg
     1741 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
     1801 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctccttcgg gctgtaatta
     1861 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct
     1921 ccgggagggc cctttgtgcg gggggagcgg ctcggggctg tccgcggggg gacggctgcc
     1981 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag
     2041 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
     2101 ttattgtgct gtctcatcat tttggcaaag aattggatcg gaccgaaatt aatacgactc
     2161 actatagggg aattgtgagc ggataacaat tccccggagt taatccggga cctttaattc
     2221 aacccaacac aatatattat agttaaataa gaattattat caaatcattt gtatattaat
     2281 taaaatacta tactgtaaat tacattttat ttacaatcaa aggagatata ccatggcaca
     2341 ccatcaccac catcactctt ctggtaaaga aaccgctgct gcgaaatttg aacgccagca
     2401 catggactcg ccaccgcctt ctggtctggt cccccggggc agcgcaggtt ctggtacgat
     2461 tgatgacgac gacaagagtc cgggcttctc ctcaacgata tctgagctcg tggatccgaa
     2521 ttctcagatc tcggcgcgcc tgcaggtcga cggtaccggt tcgaagcttg cggccgcaca
     2581 gctgtataca cgtgcaagcc agccagaact cgccccggaa gaccccgagg atctcgagca
     2641 ccaccatcac catcaccatc actaagtgat taacctcagg tgcaggctgc ctatcagaag
     2701 gtggtggctg gtgtggaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     2761 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     2821 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     2881 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     2941 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     3001 gctcacctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     3061 gggaatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa
     3121 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataag
     3181 cttgccacaa cccgtaccaa agatggatag atccggaaag cctgaactca ccgcgacgtc
     3241 tgttgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga
     3301 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt
     3361 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc
     3421 cgcgctcccg attccggaag tgcttgacat tggggaattt agcgagagcc tgacctattg
     3481 catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
     3541 tgttctgcaa ccggtcgcgg aggctatgga tgcgatcgct gcggccgatc ttagccagac
     3601 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
     3661 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
     3721 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
     3781 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
     3841 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
     3901 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
     3961 gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg
     4021 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
     4081 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
     4141 ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
     4201 ccgacgcccc agcactcgtc cgagggcaaa ggaatagagt agatgccgac cgaacaagag
     4261 ctgatttcga gaacgcctca gccagcaact cgcgcgagcc tagcaaggca aatgcgagag
     4321 aacggcctta cgcttggtgg cacagttctc gtccacagtt cgctaaccaa tgccctggct
     4381 cacaaatacc actgagatcg atctttttcc ctctgccaaa aattatgggg acatcatgaa
     4441 gccccttgag catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg
     4501 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca
     4561 tcagaatgag tatttggttt agagtttggc aacatatgcc catatgtaac tagcataacc
     4621 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaagcatgcg gaggaaattc
     4681 tccttgaagt ttccctggtg ttcaaagtaa aggagtttgc accagacgca cctctgttca
     4741 ctggtccggc gtattaaaac acgatacatt gttattagta catttattaa gcgctagatt
     4801 ctgtgcgttg ttgatttaca gacaattgtt gtacgtattt taataattca ttaaatttat
     4861 aatctttagg gtggtatgtt agagcgaaaa tcaaatgatt ttcagcgtct ttatatctga
     4921 atttaaatat taaatcctca atagatttgt aaaataggtt tcgattagtt tcaaacaagg
     4981 gttgtttttc cgaaccgatg gctggactat ctaatggatt ttcgctcaac gccacaaaac
     5041 ttgccaaatc ttgtagcagc aatctagctt tgtcgatatt cgtttgtgtt ttgttttgta
     5101 ataaaggttc gacgtcgttc aaaatattat gcgcttttgt atttctttca tcactgtcgt
     5161 tagtgtacaa ttgactcgac gtaaacacgt taaatagagc ttggacatat ttaacatcgg
     5221 gcgtgttagc tttattaggc cgattatcgt cgtcgtccca accctcgtcg ttagaagttg
     5281 cttccgaaga cgattttgcc atagccacac gacgcctatt aattgtgtcg gctaacacgt
     5341 ccgcgatcaa atttgtagtt gagctttttg gaattgcgat cgcataactt cgtatagcat
     5401 acattatacg aagttataag ctcggaacgc tgcgctcggt cgttcggctg cggcgagcgg
     5461 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5521 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5581 cgtttttcca taggctccgc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5641 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5701 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5761 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5821 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5881 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5941 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6001 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6061 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6121 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6181 tttgttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6241 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6301 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6361 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6421 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6481 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6541 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6601 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6661 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6721 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6781 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6841 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6901 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6961 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7021 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7081 agggttattg tctcatgtcc gcgcgtt
//
