LOCUS       sequence-313765-        6580 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 6580)
  AUTHORS   Yan W, Markegard E, Dharmaiah S, Urisman A, Drew M, Esposito D, 
            Scheffzek K, Nissley DV, McCormick F, Simanshu DK
  TITLE     Structural Insights into the SPRED1-Neurofibromin-KRAS Complex and 
            Disruption of SPRED1-Neurofibromin Interaction by Oncogenic EGFR.
  JOURNAL   Cell Rep. 2020 Jul 21;32(3):107909. doi: 
            10.1016/j.celrep.2020.107909.
  PUBMED    32697994
REFERENCE   2  (bases 1 to 6580)
  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: "Cell Rep.";
            date: "2020-07-21"; volume: "32(3):107909. doi"; pages: " 
            10.1016/j.celrep.2020.107909"
COMMENT     Sequence Label: pDest-635
FEATURES             Location/Qualifiers
     source          1..6580
                     /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             152..169
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    182..306
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(331..336,337..354,355..361)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             415..1074
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(481..500)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1395..1700
                     /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     1579..1598
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1741..1865)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    2023..2157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2060..2079)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2114..2133
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2186..2351
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2535..2989
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2622..2641)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2831..2852
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3015..3119
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3120..3188,3189..3980)
                     /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(3338..3357)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4151..4739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4640..4659
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4893..4910
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  5044..5268
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(5335..5868)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(6057..6062,6063..6079,6080..6085))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(6448..6557)
                     /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 gatctggatc ccaccatcac catcaccata gatcctcgat
      181 cacaagtttg tacaaaaaag ctgaacgaga aacgtaaaat gatataaata tcaatatatt
      241 aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc
      301 actatggcgg ccgcattagg caccccaggc tttacacttt atgcttccgg ctcgtataat
      361 gtgtggattt tgagttagga tccgtcgaga ttttcaggag ctaaggaagc taaaatggag
      421 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
      481 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
      541 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
      601 cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg
      661 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
      721 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
      781 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
      841 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
      901 atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag
      961 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
     1021 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaaacgcgt
     1081 ggatccggct tactaaaagc cagataacag tatgcgtatt tgcgcgcacc ggtgctagcg
     1141 tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca gcgtattaca gtgacagttg
     1201 acagcgacag ctatcagttg ctcaaggcat atatgatgtc aatatctccg gtctggtaag
     1261 cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca
     1321 ggaagggatg gctgaggtcg cccggtttat tgaaatgaac ggctcttttg ctgacgagaa
     1381 caggggctgg tgaaatgcag tttaaggttt acacctataa aagagagagc cgttatcgtc
     1441 tgtttgtgga tgtacagagt gatattattg acacgcccgg gcgacggatg gtgatccccc
     1501 tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata
     1561 tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta
     1621 tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc
     1681 tgatgttctg gggaatataa atgtcaggct cccttataca cagccagtct gcaggtcgac
     1741 catagtgact ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc
     1801 taatttaata tattgatatt tatatcattt tacgtttctc gttcagcttt cttgtacaaa
     1861 gtggtgataa ttaattaaga tcgagctagc tagaagcttg tcgagaagta ctagaggatc
     1921 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     1981 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     2041 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     2101 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatctga
     2161 tcactgcttg agcctaggag atccgaacca gataagtgaa atctagttcc aaactatttt
     2221 gtcattttta attttcgtat tagcttacga cgctacaccc agttcccatc tattttgtca
     2281 ctcttcccta aataatcctt aaaaactcca tttccacccc tcccagttcc caactatttt
     2341 gtccgcccac agcggggcat ttttcttcct gttatgtttt taatcaaaca tcctgccaac
     2401 tccatgtgac aaaccgtcat cttcggctac tttttctctg tcacagaatg aaaatttttc
     2461 tgtcatctct tcgttattaa tgtttgtaat tgactgaata tcaacgctta tttgcagcct
     2521 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2581 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2641 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2701 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     2761 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2821 gttcttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     2881 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     2941 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     3001 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3061 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3121 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3181 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3241 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3301 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3361 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3421 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3481 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3541 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3601 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3661 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3721 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3781 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3841 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3901 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3961 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4021 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4081 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4141 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4201 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4261 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4321 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4381 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4441 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4501 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4561 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4621 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4681 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4741 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4801 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     4861 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     4921 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatagacc
     4981 agccgcgtaa cctggcaaaa tcggttacgg ttgagtaata aatggatgcc ctgcgtaagc
     5041 gggtgtgggc ggacaataaa gtcttaaact gaacaaaata gatctaaact atgacaataa
     5101 agtcttaaac tagacagaat agttgtaaac tgaaatcagt ccagttatgc tgtgaaaaag
     5161 catactggac ttttgttatg gctaaagcaa actcttcatt ttctgaagtg caaattgccc
     5221 gtcgtattaa agaggggcgt ggccaagggc atggtaaaga ctatattcgc ggcgttgtga
     5281 caatttaccg aacaactccg cggccgggaa gccgatctcg gcttgaacga attgttaggt
     5341 ggcggtactt gggtcgatat caaagtgcat cacttcttcc cgtatgccca actttgtata
     5401 gagagccact gcgggatcgt caccgtaatc tgcttgcacg tagatcacat aagcaccaag
     5461 cgcgttggcc tcatgcttga ggagattgat gagcgcggtg gcaatgccct gcctccggtg
     5521 ctcgccggag actgcgagat catagatata gatctcacta cgcggctgct caaacttggg
     5581 cagaacgtaa gccgcgagag cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat
     5641 gaatgtctta ctacggagca agttcccgag gtaatcggag tccggctgat gttgggagta
     5701 ggtggctacg tctccgaact cacgaccgaa aagatcaaga gcagcccgca tggatttgac
     5761 ttggtcaggg ccgagcctac atgtgcgaat gatgcccata cttgagccac ctaactttgt
     5821 tttagggcga ctgccctgct gcgtaacatc gttgctgctg cgtaacatcg ttgctgctcc
     5881 ataacatcaa acatcgaccc acggcgtaac gcgcttgctg cttggatgcc cgaggcatag
     5941 actgtacaaa aaaacagtca taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc
     6001 tgcgttcggt caaggttctg gaccagttgc gtgagcgcat acgctacttg cattacagtt
     6061 tacgaaccga acaggcttat gtcaactggg ttcgtgcctt catccgtttc cacggtgtgc
     6121 gtcacccggc aaccttgggc agcagcgaag tcgaggcatt tctgtcctgg ctggcgaacg
     6181 agcgcaaggt ttcggtctcc acgcatcgtc aggcattggc ggccttgctg ttcttctacg
     6241 gcaaggtgct gtgcacggat ctgcccttgc ttcaggagat cggtagacct cggccgtcgc
     6301 ggcgcttgcc ggtggtgctg accccggatg aagtggttcg catcctcggt tttctggaag
     6361 gcgagcatcg tttgttcgcc caggactcta gctatagttc tagtggttgg ctactcgaga
     6421 tcccgggtga tcaagtcttc gtcgagtgat tgtaaataaa atgtaattta cagtatagta
     6481 ttttaattaa tatacaaatg atttgataat aattcttatt taactataat atattgtgtt
     6541 gggttgaatt aaaggtccgt atactccgga atattaatag
//
