LOCUS       pAcGHLT_B.        8755 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Baculovirus transfer vector for expressing proteins with an 
            N-terminal GST-6xHis-PKA-thrombin cassette. For other reading 
            frames, use pAcGHLT A or pAcGHLT C.
ACCESSION   .
VERSION     .
KEYWORDS    pAcGHLT B.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8755)
  AUTHORS   BD Biosciences
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8755)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             761..1390
                     /gene="LEF-2"
                     /label=LEF-2
                     /note="Primase-associated factor LEF-2 from Autographa 
                     californica nuclear polyhedrosis virus. Accession#: P41418"
     misc_recomb     1269..2096
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     promoter        2100..2191
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     CDS             2206..2859
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
     CDS             2875..2892
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             2899..2916
                     /codon_start=1
                     /product="recognition site for phosphorylation by protein
                     kinase A"
                     /label=recognition site for phosphorylation by protein
                     /note="PKA site"
                     /translation="RRASVA"
     CDS             2926..2943
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     misc_feature    2966..3027
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     misc_recomb     3047..4505
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     primer_bind     complement(5649..5665)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5878..6333
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6615..6719
                     /label=AmpR promoter
     CDS             6720..7577
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      7751..8339
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8627..8648
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8663..8693
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8701..8717
                     /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)."
     primer_bind     8725..8741
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tcgagcagtt cgttgacgcc ttcctccgtg tggccgaaca cgtcgagcgg gtggtcgatg
       61 accagcggcg tgccgcacgc gacgcacaag tatctgtaca ccgaatgatc gtcgggcgaa
      121 ggcacgtcgg cctccaagtg gcaatattgg caaattcgaa aatatataca gttgggttgt
      181 ttgcgcatat ctatcgtggc gttgggcatg tacgtccgaa cgttgatttg catgcaagcc
      241 gaaattaaat cattgcgatt agtgcgatta aaacgttgta catcctcgct tttaatcatg
      301 ccgtcgatta aatcgcgcaa tcgagtcaag tgatcaaagt gtggaataat gttttctttg
      361 tattcccgag tcaagcgcag cgcgtatttt aacaaactag ccatcttgta agttagtttc
      421 atttaatgca actttatcca ataatatatt atgtatcgca cgtcaagaat taacaatgcg
      481 cccgttgtcg catctcaaca cgactatgat agagatcaaa taaagcgcga attaaatagc
      541 ttgcgacgca acgtgcacga tctgtgcacg cgttccggca cgagctttga ttgtaataag
      601 tttttacgaa gcgatgacat gacccccgta gtgacaacga tcacgcccaa aagaactgcc
      661 gactacaaaa ttaccgagta tgtcggtgac gttaaaacta ttaagccatc caatcgaccg
      721 ttagtcgaat caggaccgct ggtgcgagaa gccgcgaagt atggcgaatg catcgtataa
      781 cgtgtggagt ccgctcatta gagcgtcatg tttagacaag aaagctacat atttaattga
      841 tcccgatgat tttattgata aattgaccct aactccatac acggtattct acaatggcgg
      901 ggttttggtc aaaatttccg gactgcgatt gtacatgctg ttaacggctc cgcccactat
      961 taatgaaatt aaaaattcca attttaaaaa acgcagcaag agaaacattt gtatgaaaga
     1021 atgcgtagaa ggaaagaaaa atgtcgtcga catgctgaac aacaagatta atatgcctcc
     1081 gtgtataaaa aaaatattga acgatttgaa agaaaacaat gtaccgcgcg gcggtatgta
     1141 caggaagagg tttatactaa actgttacat tgcaaacgtg gtttcgtgtg ccaagtgtga
     1201 aaaccgatgt ttaatcaagg ctctgacgca tttctacaac cacgactcca agtgtgtggg
     1261 tgaagtcatg catcttttaa tcaaatccca agatgtgtat aaaccaccaa actgccaaaa
     1321 aatgaaaact gtcgacaagc tctgtccgtt tgctggcaac tgcaagggtc tcaatcctat
     1381 ttgtaattat tgaataataa aacaattata aatgctaaat ttgtttttta ttaacgatac
     1441 aaaccaaacg caacaagaac atttgtagta ttatctataa ttgaaaacgc gtagttataa
     1501 tcgctgaggt aatatttaaa atcattttca aatgattcac agttaatttg cgacaatata
     1561 attttatttt cacataaact agacgccttg tcgtcttctt cttcgtattc cttctctttt
     1621 tcatttttct cctcataaaa attaacatag ttattatcgt atccatatat gtatctatcg
     1681 tatagagtaa attttttgtt gtcataaata tatatgtctt ttttaatggg gtgtatagta
     1741 ccgctgcgca tagtttttct gtaatttaca acagtgctat tttctggtag ttcttcggag
     1801 tgtgttgctt taattattaa atttatataa tcaatgaatt tgggatcgtc ggttttgtac
     1861 aatatgttgc cggcatagta cgcagcttct tctagttcaa ttacaccatt ttttagcagc
     1921 accggattaa cataactttc caaaatgttg tacgaaccgt taaacaaaaa cagttcacct
     1981 cccttttcta tactattgtc tgcgagcagt tgtttgttgt taaaaataac agccattgta
     2041 atgagacgca caaactaata tcacaaactg gaaatgtcta tcaatatata gttgctgata
     2101 tcatggagat aattaaaatg ataaccatct cgcaaataaa taagtatttt actgttttcg
     2161 taacagtttt gtaataaaaa aacctataaa tacggatctg tattcatgtc ccctatacta
     2221 ggttattgga aaattaaggg ccttgtgcaa cccactcgac ttcttttgga atatcttgaa
     2281 gaaaaatatg aagagcattt gtatgagcgc gatgaaggtg ataaatggcg aaacaaaaag
     2341 tttgaattgg gtttggagtt tcccaatctt ccttattata ttgatggtga tgttaaatta
     2401 acacagtcta tggccatcat acgttatata gctgacaagc acaacatgtt gggtggttgt
     2461 ccaaaagagc gtgcagagat ttcaatgctt gaaggagcgg ttttggatat tagatacggt
     2521 gtttcgagaa ttgcatatag taaagacttt gaaactctca aagttgattt tcttagcaag
     2581 ctacctgaaa tgctgaaaat gttcgaagat cgtttatgtc ataaaacata tttaaatggt
     2641 gatcatgtaa cccatcctga cttcatgttg tatgacgctc ttgatgttgt tttatacatg
     2701 gacccaatgt gcctggatgc gttcccaaaa ttagtttgtt ttaaaaaacg tattgaagct
     2761 atcccacaaa ttgataagta cttgaaatcc agcaagtata tagcatggcc tttgcagggc
     2821 tggcaagcca cgtttggtgg tggcgaccat cctccaaaat cggatccgat gggacatcat
     2881 catcatcatc acggaaggag aagggccagt gttgcggcgg gaattttggt ccctcgtgga
     2941 agcccaggac tcgatggcat atatgctcga ggaattcagg cctccatggg agctcgcggc
     3001 cgcctgcagg gtacccccgg gagatctgta ccgactctgc tgaagaggag gaaattctcc
     3061 ttgaagtttc cctggtgttc aaagtaaagg agtttgcacc agacgcacct ctgttcactg
     3121 gtccggcgta ttaaaacacg atacattgtt attagtacat ttattaagcg ctagattctg
     3181 tgcgttgttg atttacagac aattgttgta cgtattttaa taattcatta aatttataat
     3241 ctttagggtg gtatgttaga gcgaaaatca aatgattttc agcgtcttta tatctgaatt
     3301 taaatattaa atcctcaata gatttgtaaa ataggtttcg attagtttca aacaagggtt
     3361 gtttttccga accgatggct ggactatcta atggattttc gctcaacgcc acaaaacttg
     3421 ccaaatcttg tagcagcaat ctagctttgt cgatattcgt ttgtgttttg ttttgtaata
     3481 aaggttcgac gtcgttcaaa atattatgcg cttttgtatt tctttcatca ctgtcgttag
     3541 tgtacaattg actcgacgta aacacgttaa ataaagcttg gacatattta acatcgggcg
     3601 tgttagcttt attaggccga ttatcgtcgt cgtcccaacc ctcgtcgtta gaagttgctt
     3661 ccgaagacga ttttgccata gccacacgac gcctattaat tgtgtcggct aacacgtccg
     3721 cgatcaaatt tgtagttgag ctttttggaa ttatttctga ttgcgggcgt ttttgggcgg
     3781 gtttcaatct aactgtgccc gattttaatt cagacaacac gttagaaagc gatggtgcag
     3841 gcggtggtaa catttcagac ggcaaatcta ctaatggcgg cggtggtgga gctgatgata
     3901 aatctaccat cggtggaggc gcaggcgggg ctggcggcgg aggcggaggc ggaggtggtg
     3961 gcggtgatgc agacggcggt ttaggctcaa atgtctcttt aggcaacaca gtcggcacct
     4021 caactattgt actggtttcg ggcgccgttt ttggtttgac cggtctgaga cgagtgcgat
     4081 ttttttcgtt tctaatagct tccaacaatt gttgtctgtc gtctaaaggt gcagcgggtt
     4141 gaggttccgt cggcattggt ggagcgggcg gcaattcaga catcgatggt ggtggtggtg
     4201 gtggaggcgc tggaatgtta ggcacgggag aaggtggtgg cggcggtgcc gccggtataa
     4261 tttgttctgg tttagtttgt tcgcgcacga ttgtgggcac cggcgcaggc gccgctggct
     4321 gcacaacgga aggtcgtctg cttcgaggca gcgcttgggg tggtggcaat tcaatattat
     4381 aattggaata caaatcgtaa aaatctgcta taagcattgt aatttcgcta tcgtttaccg
     4441 tgccgatatt taacaaccgc tcaatgtaag caattgtatt gtaaagagat tgtctcaagc
     4501 tcggatcgat cccgcacgcc gataacaagc cttttcattt ttactacagc attgtagtgg
     4561 cgagacactt cgctgtcgtc gacgtacatg tatgctttgt tgtcaaaaac gtcgttggca
     4621 agctttaaaa tatttaaaag aacatctctg ttcagcacca ctgtgttgtc gtaaatgttg
     4681 tttttgataa tttgcgcttc cgcagtatcg acacgttcaa aaaattgatg cgcatcaatt
     4741 ttgttgttcc tattattgaa taaataagat tgtacagatt catatctacg attcgtcatg
     4801 gccaccacaa atgctacgct gcaaacgctg gtacaatttt acgaaaactg caaaaacgtc
     4861 aaaactcggt ataaaataat caacgggcgc tttggcaaaa tatctatttt atcgcacaag
     4921 cccactagca aattgtattt gcagaaaaca atttcggcgc acaattttaa cgctgacgaa
     4981 ataaaagttc accagttaat gagcgaccac ccaaatttta taaaaatcta ttttaatcac
     5041 ggttccatca acaaccaagt gatcgtgatg gactacattg actgtcccga tttatttgaa
     5101 acactacaaa ttaaaggcga gctttcgtac caacttgtta gcaatattat tagacagctg
     5161 tgtgaagcgc tcaacgattt gcacaagcac aatttcatac acaacgacat aaaactcgaa
     5221 aatgtcttat atttcgaagc acttgatcgc gtgtatgttt gcgattacgg attgtgcaaa
     5281 cacgaaaact cacttagcgt gcacgacggc acgttggagt attttagtcc ggaaaaaatt
     5341 cgacacacaa ctatgcacgt ttcgtttgac tggtacgcgg cgtgttaaca tacaagttgc
     5401 taaccggcgg ccgacaccca tttgaaaaaa gcgaagacga aatgttggac ttgaatagca
     5461 tgaagcgtcg tcagcaatac aatgacattg gcgttttaaa acacgttcgt aacgttaacg
     5521 ctcgtgactt tgtgtactgc ctaacaagat acaacataga ttgtagactc acaaattaca
     5581 aacaaattat aaaacatgag tttttgtcgt aaaaatgcca cttgttttac gagtagaatt
     5641 agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     5701 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     5761 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
     5821 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     5881 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     5941 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     6001 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     6061 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     6121 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     6181 tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag
     6241 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     6301 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     6361 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     6421 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     6481 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     6541 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     6601 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     6661 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     6721 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     6781 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     6841 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     6901 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     6961 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     7021 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     7081 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     7141 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     7201 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     7261 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     7321 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     7381 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     7441 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     7501 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     7561 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     7621 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     7681 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     7741 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     7801 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     7861 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     7921 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7981 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     8041 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     8101 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     8161 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     8221 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     8281 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     8341 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     8401 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     8461 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     8521 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     8581 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     8641 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     8701 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gaatt
//
