LOCUS       sequence-313186-        5303 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculovirus protein expression of His6-tev-Hs.SPRED1(13-125).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5303)
  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 5303)
  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: R725-X09-635: His6-tev-Hs.SPRED1(13-125)
FEATURES             Location/Qualifiers
     source          1..5303
                     /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..206
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             206..226
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     protein_bind    complement(564..588)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    746..880
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(783..802)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     837..856
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  909..1074
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      1258..1712
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1345..1364)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1554..1575
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1738..1842
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1843..1911,1912..2703)
                     /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(2061..2080)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2874..3462
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3363..3382
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3616..3633
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  3767..3991
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4058..4591)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4780..4785,4786..4802,4803..4808))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(5171..5280)
                     /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 ttggcgaaaa cctgtacttc caaagttatg cacgagtgcg
      241 agctgtggtg atgacccgag atgactcaag tggtggatgg ttaccacttg gagggagtgg
      301 actaagcagc gtcactgtct tcaaagtccc tcatcaggaa gagaatggct gtgctgactt
      361 ttttatccgt ggagagcgac tcagggacaa aatggtggtt ttggaatgta tgcttaaaaa
      421 agacctcatt tataataagg tcactccaac atttcaccac tggaagattg atgacaaaaa
      481 gtttggtctt acgtttcaaa gtcctgctga tgctagggct tttgatagag gtatccgaag
      541 agctatagag gatatttctc aataatcaac tttcttgtac aaagtggtga taattaatta
      601 agatcgagct agctagaagc ttgtcgagaa gtactagagg atcataatca gccataccac
      661 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
      721 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
      781 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
      841 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc tgatcactgc ttgagcctag
      901 gagatccgaa ccagataagt gaaatctagt tccaaactat tttgtcattt ttaattttcg
      961 tattagctta cgacgctaca cccagttccc atctattttg tcactcttcc ctaaataatc
     1021 cttaaaaact ccatttccac ccctcccagt tcccaactat tttgtccgcc cacagcgggg
     1081 catttttctt cctgttatgt ttttaatcaa acatcctgcc aactccatgt gacaaaccgt
     1141 catcttcggc tactttttct ctgtcacaga atgaaaattt ttctgtcatc tcttcgttat
     1201 taatgtttgt aattgactga atatcaacgc ttatttgcag cctgaatggc gaatgggacg
     1261 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1321 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     1381 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1441 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     1501 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttctta atagtggact
     1561 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     1621 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     1681 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     1741 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1801 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1861 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1921 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1981 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     2041 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     2101 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     2161 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     2221 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     2281 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2341 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     2401 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2461 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2521 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2581 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     2641 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     2701 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2761 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2821 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2881 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2941 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3001 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     3061 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3121 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3181 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3241 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3301 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3361 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3421 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3481 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3541 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3601 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     3661 tttctcctta cgcatctgtg cggtatttca caccgcatag accagccgcg taacctggca
     3721 aaatcggtta cggttgagta ataaatggat gccctgcgta agcgggtgtg ggcggacaat
     3781 aaagtcttaa actgaacaaa atagatctaa actatgacaa taaagtctta aactagacag
     3841 aatagttgta aactgaaatc agtccagtta tgctgtgaaa aagcatactg gacttttgtt
     3901 atggctaaag caaactcttc attttctgaa gtgcaaattg cccgtcgtat taaagagggg
     3961 cgtggccaag ggcatggtaa agactatatt cgcggcgttg tgacaattta ccgaacaact
     4021 ccgcggccgg gaagccgatc tcggcttgaa cgaattgtta ggtggcggta cttgggtcga
     4081 tatcaaagtg catcacttct tcccgtatgc ccaactttgt atagagagcc actgcgggat
     4141 cgtcaccgta atctgcttgc acgtagatca cataagcacc aagcgcgttg gcctcatgct
     4201 tgaggagatt gatgagcgcg gtggcaatgc cctgcctccg gtgctcgccg gagactgcga
     4261 gatcatagat atagatctca ctacgcggct gctcaaactt gggcagaacg taagccgcga
     4321 gagcgccaac aaccgcttct tggtcgaagg cagcaagcgc gatgaatgtc ttactacgga
     4381 gcaagttccc gaggtaatcg gagtccggct gatgttggga gtaggtggct acgtctccga
     4441 actcacgacc gaaaagatca agagcagccc gcatggattt gacttggtca gggccgagcc
     4501 tacatgtgcg aatgatgccc atacttgagc cacctaactt tgttttaggg cgactgccct
     4561 gctgcgtaac atcgttgctg ctgcgtaaca tcgttgctgc tccataacat caaacatcga
     4621 cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac aaaaaaacag
     4681 tcataacaag ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt
     4741 ctggaccagt tgcgtgagcg catacgctac ttgcattaca gtttacgaac cgaacaggct
     4801 tatgtcaact gggttcgtgc cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg
     4861 ggcagcagcg aagtcgaggc atttctgtcc tggctggcga acgagcgcaa ggtttcggtc
     4921 tccacgcatc gtcaggcatt ggcggccttg ctgttcttct acggcaaggt gctgtgcacg
     4981 gatctgccct tgcttcagga gatcggtaga cctcggccgt cgcggcgctt gccggtggtg
     5041 ctgaccccgg atgaagtggt tcgcatcctc ggttttctgg aaggcgagca tcgtttgttc
     5101 gcccaggact ctagctatag ttctagtggt tggctactcg agatcccggg tgatcaagtc
     5161 ttcgtcgagt gattgtaaat aaaatgtaat ttacagtata gtattttaat taatatacaa
     5221 atgatttgat aataattctt atttaactat aatatattgt gttgggttga attaaaggtc
     5281 cgtatactcc ggaatattaa tag
//
