LOCUS       sequence-313763-        7689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Destination vector for protein expression in baculovirus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7689)
  AUTHORS   Gillette WK, Esposito D, Abreu Blanco M, Alexander P, Bindu L, 
            Bittner C, Chertov O, Frank PH, Grose C, Jones JE, Meng Z, Perkins 
            S, Van Q, Ghirlando R, Fivash M, Nissley DV, McCormick F, 
            Holderfield M, Stephen AG
  TITLE     Farnesylated and methylated KRAS4b: high yield production of protein
            suitable for biophysical studies of prenylated protein-lipid 
            interactions.
  JOURNAL   Sci Rep. 2015 Nov 2;5:15916. doi: 10.1038/srep15916.
  PUBMED    26522388
REFERENCE   2  (bases 1 to 7689)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2015-11-2"; volume: "5:15916. doi"; pages: " 
            10.1038/srep15916"
COMMENT     Sequence Label: pDest-636
FEATURES             Location/Qualifiers
     source          1..7689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..92
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     17..34
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             146..163
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             167..1264
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1121..1130
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1238..1261
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     protein_bind    1283..1407
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1432..1437,1438..1455,1456..1462)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1516..2175
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1582..1601)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2517..2822
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2701..2720
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2863..2987)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    3132..3266
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3169..3188)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3223..3242
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3295..3460
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3644..4098
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3731..3750)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3940..3961
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4124..4228
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4229..4297,4298..5089)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4447..4466)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5260..5848
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5749..5768
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6002..6019
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6153..6377
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6444..6977)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7166..7171,7172..7188,7189..7194))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(7557..7666)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
ORIGIN
        1 atcatggaga taattaaaat gataaccatc tcgcaaataa ataagtattt tactgttttc
       61 gtaacagttt tgtaataaaa aaacctataa atattccgga ttattcatac cgtcccacca
      121 tcgggcgcgg atcaccatgg gatcccatca ccatcaccat cacggcaaaa tcgaagaagg
      181 taaactggta atctggatta acggcgataa aggctataac ggtctcgctg aagtcggtaa
      241 gaaattcgag aaagataccg gaattaaagt caccgttgag catccggata aactggaaga
      301 gaaattccca caggttgcgg caactggcga tggccctgac attatcttct gggcacacga
      361 ccgctttggt ggctacgctc aatctggcct gttggctgaa atcaccccgg acaaagcgtt
      421 ccaggacaag ctgtatccgt ttacctggga tgccgtacgt tacaacggca agctgattgc
      481 ttacccgatc gctgttgaag cgttatcgct gatttataac aaagatctgc tgccgaaccc
      541 gccaaaaacc tgggaagaga tcccggcgct ggataaagaa ctgaaagcga aaggtaagag
      601 cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg ccgctgattg ctgctgacgg
      661 gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa
      721 cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg attaaaaaca aacacatgaa
      781 tgcagacacc gattactcca tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac
      841 catcaacggc ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt atggtgtaac
      901 ggtactgccg accttcaagg gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg
      961 tattaacgct gccagtccga acaaagagct ggcaaaagag ttcctcgaaa actatctgct
     1021 gactgatgaa ggtctggaag cggttaataa agacaaaccg ctgggtgccg tagcgctgaa
     1081 gtcttacgag gaagagttgg cgaaagatcc acgtattgcc gccaccatgg aaaacgccca
     1141 gaaaggtgaa atcatgccga acatcccgca gatgtccgct ttctggtatg ccgtgcgtac
     1201 tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat gaagccctga aagacgcgca
     1261 gactaattcg ggatcctcga tcacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa
     1321 tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta cataatactg
     1381 taaaacacaa catatccagt cactatggcg gccgcattag gcaccccagg ctttacactt
     1441 tatgcttccg gctcgtataa tgtgtggatt ttgagttagg atccgtcgag attttcagga
     1501 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
     1561 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
     1621 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
     1681 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg
     1741 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
     1801 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
     1861 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
     1921 aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt
     1981 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
     2041 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
     2101 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
     2161 cagggcgggg cgtaaacgcg tggatccggc ttactaaaag ccagataaca gtatgcgtat
     2221 ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca
     2281 aaaagaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt
     2341 tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga
     2401 agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt
     2461 cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct ggtgaaatgc
     2521 agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga
     2581 gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc
     2641 tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc
     2701 gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg
     2761 atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat
     2821 aaatgtcagg ctcccttata cacagccagt ctgcaggtcg accatagtga ctggatatgt
     2881 tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata
     2941 tttatatcat tttacgtttc tcgttcagct ttcttgtaca aagtggtgat cgagctagct
     3001 agaagcttgt cgagaagtac tagaggatca taatcagcca taccacattt gtagaggttt
     3061 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     3121 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3181 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3241 tcaatgtatc ttatcatgtc tggatctgat cactgcttga gcctaggaga tccgaaccag
     3301 ataagtgaaa tctagttcca aactattttg tcatttttaa ttttcgtatt agcttacgac
     3361 gctacaccca gttcccatct attttgtcac tcttccctaa ataatcctta aaaactccat
     3421 ttccacccct cccagttccc aactattttg tccgcccaca gcggggcatt tttcttcctg
     3481 ttatgttttt aatcaaacat cctgccaact ccatgtgaca aaccgtcatc ttcggctact
     3541 ttttctctgt cacagaatga aaatttttct gtcatctctt cgttattaat gtttgtaatt
     3601 gactgaatat caacgcttat ttgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
     3661 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3721 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3781 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3841 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3901 gtttttcgcc ctttgacgtt ggagtccacg ttcttaatag tggactcttg ttccaaactg
     3961 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4021 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4081 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4141 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4201 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4261 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4321 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4381 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4441 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4501 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4561 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4621 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4681 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4741 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4801 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4861 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4921 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4981 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5041 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5101 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5161 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5221 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5281 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5341 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5401 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5461 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5521 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5581 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5641 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5701 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5761 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5821 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5881 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5941 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6001 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     6061 tctgtgcggt atttcacacc gcatagacca gccgcgtaac ctggcaaaat cggttacggt
     6121 tgagtaataa atggatgccc tgcgtaagcg ggtgtgggcg gacaataaag tcttaaactg
     6181 aacaaaatag atctaaacta tgacaataaa gtcttaaact agacagaata gttgtaaact
     6241 gaaatcagtc cagttatgct gtgaaaaagc atactggact tttgttatgg ctaaagcaaa
     6301 ctcttcattt tctgaagtgc aaattgcccg tcgtattaaa gaggggcgtg gccaagggca
     6361 tggtaaagac tatattcgcg gcgttgtgac aatttaccga acaactccgc ggccgggaag
     6421 ccgatctcgg cttgaacgaa ttgttaggtg gcggtacttg ggtcgatatc aaagtgcatc
     6481 acttcttccc gtatgcccaa ctttgtatag agagccactg cgggatcgtc accgtaatct
     6541 gcttgcacgt agatcacata agcaccaagc gcgttggcct catgcttgag gagattgatg
     6601 agcgcggtgg caatgccctg cctccggtgc tcgccggaga ctgcgagatc atagatatag
     6661 atctcactac gcggctgctc aaacttgggc agaacgtaag ccgcgagagc gccaacaacc
     6721 gcttcttggt cgaaggcagc aagcgcgatg aatgtcttac tacggagcaa gttcccgagg
     6781 taatcggagt ccggctgatg ttgggagtag gtggctacgt ctccgaactc acgaccgaaa
     6841 agatcaagag cagcccgcat ggatttgact tggtcagggc cgagcctaca tgtgcgaatg
     6901 atgcccatac ttgagccacc taactttgtt ttagggcgac tgccctgctg cgtaacatcg
     6961 ttgctgctgc gtaacatcgt tgctgctcca taacatcaaa catcgaccca cggcgtaacg
     7021 cgcttgctgc ttggatgccc gaggcataga ctgtacaaaa aaacagtcat aacaagccat
     7081 gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc aaggttctgg accagttgcg
     7141 tgagcgcata cgctacttgc attacagttt acgaaccgaa caggcttatg tcaactgggt
     7201 tcgtgccttc atccgtttcc acggtgtgcg tcacccggca accttgggca gcagcgaagt
     7261 cgaggcattt ctgtcctggc tggcgaacga gcgcaaggtt tcggtctcca cgcatcgtca
     7321 ggcattggcg gccttgctgt tcttctacgg caaggtgctg tgcacggatc tgcccttgct
     7381 tcaggagatc ggtagacctc ggccgtcgcg gcgcttgccg gtggtgctga ccccggatga
     7441 agtggttcgc atcctcggtt ttctggaagg cgagcatcgt ttgttcgccc aggactctag
     7501 ctatagttct agtggttggc tactcgagat cccgggtgat caagtcttcg tcgagtgatt
     7561 gtaaataaaa tgtaatttac agtatagtat tttaattaat atacaaatga tttgataata
     7621 attcttattt aactataata tattgtgttg ggttgaatta aaggtccgta tactccggaa
     7681 tattaatag
//
