LOCUS       sequence-218652-        7851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of His-p97 in insect cells (Baculovirus system).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7851)
  AUTHORS   Weith M, Seiler J, van den Boom J, Kracht M, Hulsmann J, Primorac I,
            Del Pino Garcia J, Kaschani F, Kaiser M, Musacchio A, Bollen M, 
            Meyer H
  TITLE     Ubiquitin-Independent Disassembly by a p97 AAA-ATPase Complex Drives
            PP1 Holoenzyme Formation.
  JOURNAL   Mol Cell. 2018 Oct 11. pii: S1097-2765(18)30791-3. doi: 
            10.1016/j.molcel.2018.09.020.
  PUBMED    30344098
REFERENCE   2  (bases 1 to 7851)
  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: "Mol Cell. 
            2018 Oct 11. pii: S1097-2765(18)30791-3. doi: 
            10.1016/j.molcel.2018.09.020."
COMMENT     Sequence Label: pFL His-p97
FEATURES             Location/Qualifiers
     source          1..7851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        59..150
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     75..92
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             196..213
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2810..2944
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2847..2866)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2901..2920
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3054..3219
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3403..3857
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3490..3509)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3699..3720
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3883..3987
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3988..4056,4057..4848)
                     /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(4206..4225)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5019..5607
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5508..5527
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5761..5778
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  5912..6136
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6203..6736)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(6925..6930,6931..6947,6948..6953))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(7349..7382)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    7572..7620
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     7645..7664
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(join(7756..7851,1..14))
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
ORIGIN
        1 ttgaattaaa ggtccgtata ctagtatcga ttcgcgacct actccggaat attaatagat
       61 catggagata attaaaatga taaccatctc gcaaataaat aagtatttta ctgttttcgt
      121 aacagttttg taataaaaaa acctataaat attccggatt attcataccg tcccaccatc
      181 gggcgcggat ccatgcacca tcaccatcac catgcttctg gagccgattc aaaaggtgat
      241 gacctatcaa cagccattct caaacagaag aaccgtccca atcggttaat tgttgatgaa
      301 gccatcaatg aggacaacag tgtggtgtcc ttgtcccagc ccaagatgga tgaattgcag
      361 ttgttccgag gtgacacagt gttgctgaaa ggaaagaaga gacgagaagc tgtttgcatc
      421 gtcctttctg atgatacttg ttctgatgag aagattcgga tgaatagagt tgttcggaat
      481 aaccttcgtg tacgcctagg ggatgtcatc agcatccagc catgccctga tgtgaagtac
      541 ggcaaacgta tccatgtgct gcccattgat gacacagtgg aaggcattac tggtaatctc
      601 ttcgaggtat accttaagcc gtacttcctg gaagcgtatc gacccatccg gaaaggagac
      661 atttttcttg tccgtggtgg gatgcgtgct gtggagttca aagtggtgga aacagatcct
      721 agcccttatt gcattgttgc tccagacaca gtgatccact gcgaagggga gcctatcaaa
      781 cgagaggatg aggaagagtc cttgaatgaa gtagggtatg atgacattgg tggctgcagg
      841 aagcagctag ctcagataaa ggagatggtg gaactgcccc tgagacatcc tgccctcttt
      901 aaggcaattg gtgtgaagcc tcctagagga atcctgcttt acggacctcc tggaacagga
      961 aagaccctga ttgctcgagc tgtagcaaat gagactggag ccttcttctt cttgatcaat
     1021 ggtcctgaga tcatgagcaa attggctggt gagtctgaga gcaaccttcg taaagccttt
     1081 gaggaggctg agaagaatgc tcctgccatc atcttcattg atgagctaga tgccatcgct
     1141 cccaaaagag agaaaactca tggcgaggtg gagcggcgca ttgtatcaca gttgttgacc
     1201 ctcatggatg gcctaaagca gagggcacat gtgattgtta tggcagcaac caacagaccc
     1261 aacagcattg acccagctct acggcgattt ggtcgctttg acagggaggt agatattgga
     1321 attcctgatg ctacaggacg cttagagatt cttcagatcc ataccaagaa catgaagctg
     1381 gcagatgatg tggacctgga acaggtagcc aatgagactc acgggcatgt gggtgctgac
     1441 ttagcagccc tgtgctcaga ggctgctctg caagccatcc gcaagaagat ggatctcatt
     1501 gacctagagg atgagaccat tgatgccgag gtcatgaact ctctagcagt tactatggat
     1561 gacttccggt gggccttgag ccagagtaac ccatcagcac tgcgggaaac cgtggtagag
     1621 gtgccacagg taacctggga agacatcggg ggcctagagg atgtcaaacg tgagctacag
     1681 gagctggtcc agtatcctgt ggagcaccca gacaaattcc tgaagtttgg catgacacct
     1741 tccaagggag ttctgttcta tggacctcct ggctgtggga aaactttgtt ggccaaagcc
     1801 attgctaatg aatgccaggc caacttcatc tccatcaagg gtcctgagct gctcaccatg
     1861 tggtttgggg agtctgaggc caatgtcaga gaaatctttg acaaggcccg ccaagctgcc
     1921 ccctgtgtgc tattctttga tgagctggat tcgattgcca aggctcgtgg aggtaacatt
     1981 ggagatggtg gtggggctgc tgaccgagtc atcaaccaga tcctgacaga aatggatggc
     2041 atgtccacta aaaaaaatgt gttcatcatt ggcgctacca accggcctga catcattgat
     2101 cctgccatcc tcagacctgg ccgtcttgat cagctcatct acatcccact tcctgatgag
     2161 aagtcccgtg ttgccatcct caaggctaac ctgcgcaagt ccccagttgc caaggatgtg
     2221 gacttggagt tcctggctaa aatgactaat ggcttctctg gagctgacct gacagagatt
     2281 tgccagcgtg cttgcaagct ggccatccgt gaatccatcg agagtgagat taggcgagaa
     2341 cgagagaggc agacaaaccc atcagccatg gaggtagaag aggatgatcc agtgcctgag
     2401 atccgtcgag atcactttga agaagccatg cgctttgcgc gccgttctgt cagtgacaat
     2461 gacattcgga agtatgagat gtttgcccag acccttcagc agagtcgggg ctttggcagc
     2521 ttcagattcc cttcagggaa ccagggtgga gctggcccca gtcagggcag tggaggcggc
     2581 acaggtggca gtgtatacac agaagacaat gatgatgacc tgtatggcta agagctcact
     2641 agtcgcggcc gctttcgaat ctagagcctg cagtctcgac aagcttgtcg agaagtacta
     2701 gaggatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     2761 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     2821 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     2881 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     2941 gatctgatca ctgcttgagc ctagaagatc cggctgctaa caaagcccga aaggaagctg
     3001 agttggctgc tgccaccgct gagcaataac tatcataacc cctaggagat ccgaaccaga
     3061 taagtgaaat ctagttccaa actattttgt catttttaat tttcgtatta gcttacgacg
     3121 ctacacccag ttcccatcta ttttgtcact cttccctaaa taatccttaa aaactccatt
     3181 tccacccctc ccagttccca actattttgt ccgcccacag cggggcattt ttcttcctgt
     3241 tatgttttta atcaaacatc ctgccaactc catgtgacaa accgtcatct tcggctactt
     3301 tttctctgtc acagaatgaa aatttttctg tcatctcttc gttattaatg tttgtaattg
     3361 actgaatatc aacgcttatt tgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     3421 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3481 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3541 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3601 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3661 tttttcgccc tttgacgttg gagtccacgt tcttaatagt ggactcttgt tccaaactgg
     3721 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3781 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3841 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3901 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3961 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4021 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4081 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4141 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4201 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4261 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4321 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4381 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4441 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4501 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4561 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4621 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4681 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4741 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4801 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4861 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4921 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4981 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5041 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5101 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5161 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5221 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5281 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5341 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5401 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5461 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5521 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5581 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5641 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5701 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     5761 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     5821 ctgtgcggta tttcacaccg catagaccag ccgcgtaacc tggcaaaatc ggttacggtt
     5881 gagtaataaa tggatgccct gcgtaagcgg gtgtgggcgg acaataaagt cttaaactga
     5941 acaaaataga tctaaactat gacaataaag tcttaaacta gacagaatag ttgtaaactg
     6001 aaatcagtcc agttatgctg tgaaaaagca tactggactt ttgttatggc taaagcaaac
     6061 tcttcatttt ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg ccaagggcat
     6121 ggtaaagact atattcgcgg cgttgtgaca atttaccgaa caactccgcg gccgggaagc
     6181 cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg gtcgatatca aagtgcatca
     6241 cttcttcccg tatgcccaac tttgtataga gagccactgc gggatcgtca ccgtaatctg
     6301 cttgcacgta gatcacataa gcaccaagcg cgttggcctc atgcttgagg agattgatga
     6361 gcgcggtggc aatgccctgc ctccggtgct cgccggagac tgcgagatca tagatataga
     6421 tctcactacg cggctgctca aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg
     6481 cttcttggtc gaaggcagca agcgcgatga atgtcttact acggagcaag ttcccgaggt
     6541 aatcggagtc cggctgatgt tgggagtagg tggctacgtc tccgaactca cgaccgaaaa
     6601 gatcaagagc agcccgcatg gatttgactt ggtcagggcc gagcctacat gtgcgaatga
     6661 tgcccatact tgagccacct aactttgttt tagggcgact gccctgctgc gtaacatcgt
     6721 tgctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc
     6781 gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg
     6841 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
     6901 gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt
     6961 tgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc
     7021 gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag
     7081 gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct gcccttgctt
     7141 caggagatcg gtagacctcg gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa
     7201 gtggttcgca tcctcggttt tctggaaggc gagcatcgtt tgttcgccca ggactctagc
     7261 tatagttcta gtggttggct acagctttgt ttgtactatc aacaggttga actgctgatc
     7321 aacagatcct ctacgcggcc gcggtaccat aacttcgtat agcatacatt atacgaagtt
     7381 atctggttta aacgtacccg tagtggctat ggcagggctt gccgccccga cgttggctgc
     7441 gagccctggg ccttcacccg aacttggggg ttggggtggg gaaaaggaag aaacgcgggc
     7501 gtattggtcc caatggggtc tcggtggggt atcgacagag tgccagccct gggaccgaac
     7561 cccgcgttta tgaacaaacg acccaacacc cgtgcgtttt attctgtctt tttattgccg
     7621 tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc ctcccccatc
     7681 tcccggtacc gcatgctatg catcagctgc tagcaccatg gctcgagatc ccgggtgatc
     7741 aagtcttcgt cgagtgattg taaataaaat gtaatttaca gtatagtatt ttaattaata
     7801 tacaaatgat ttgataataa ttcttattta actataatat attgtgttgg g
//
