LOCUS       sequence-214246-        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"
     rep_origin      102..556
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(189..208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     398..419
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        582..686
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(687..755,756..1547)
                     /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(905..924)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1718..2306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2207..2226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2460..2477
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2611..2835
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2902..3435)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3624..3629,3630..3646,3647..3652))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(4048..4081)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    4271..4319
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4344..4363
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(4455..4564)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        4609..4700
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4625..4642
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4746..4763
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    7360..7494
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7397..7416)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7451..7470
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  7604..7769
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
       61 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
      121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      361 ttttcgccct ttgacgttgg agtccacgtt cttaatagtg gactcttgtt ccaaactgga
      421 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
      481 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      541 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      601 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      661 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      721 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      781 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
      841 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
      901 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
      961 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1021 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1081 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1141 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1201 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     1261 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1321 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1381 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1441 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1501 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1561 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1621 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1681 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1741 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1801 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1861 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1921 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1981 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2041 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2101 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2161 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2221 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2281 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2341 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2401 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2461 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     2521 tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct ggcaaaatcg gttacggttg
     2581 agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa
     2641 caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga
     2701 aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact
     2761 cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg
     2821 gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc
     2881 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac
     2941 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc
     3001 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag
     3061 cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat
     3121 ctcactacgc ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc caacaaccgc
     3181 ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta
     3241 atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag
     3301 atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat
     3361 gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt
     3421 gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
     3481 cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga
     3541 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     3601 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc
     3661 gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg
     3721 aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg
     3781 cattggcggc cttgctgttc ttctacggca aggtgctgtg cacggatctg cccttgcttc
     3841 aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag
     3901 tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct
     3961 atagttctag tggttggcta cagctttgtt tgtactatca acaggttgaa ctgctgatca
     4021 acagatcctc tacgcggccg cggtaccata acttcgtata gcatacatta tacgaagtta
     4081 tctggtttaa acgtacccgt agtggctatg gcagggcttg ccgccccgac gttggctgcg
     4141 agccctgggc cttcacccga acttgggggt tggggtgggg aaaaggaaga aacgcgggcg
     4201 tattggtccc aatggggtct cggtggggta tcgacagagt gccagccctg ggaccgaacc
     4261 ccgcgtttat gaacaaacga cccaacaccc gtgcgtttta ttctgtcttt ttattgccgt
     4321 catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc tcccccatct
     4381 cccggtaccg catgctatgc atcagctgct agcaccatgg ctcgagatcc cgggtgatca
     4441 agtcttcgtc gagtgattgt aaataaaatg taatttacag tatagtattt taattaatat
     4501 acaaatgatt tgataataat tcttatttaa ctataatata ttgtgttggg ttgaattaaa
     4561 ggtccgtata ctagtatcga ttcgcgacct actccggaat attaatagat catggagata
     4621 attaaaatga taaccatctc gcaaataaat aagtatttta ctgttttcgt aacagttttg
     4681 taataaaaaa acctataaat attccggatt attcataccg tcccaccatc gggcgcggat
     4741 ccatgcacca tcaccatcac catgcttctg gagccgattc aaaaggtgat gacctatcaa
     4801 cagccattct caaacagaag aaccgtccca atcggttaat tgttgatgaa gccatcaatg
     4861 aggacaacag tgtggtgtcc ttgtcccagc ccaagatgga tgaattgcag ttgttccgag
     4921 gtgacacagt gttgctgaaa ggaaagaaga gacgagaagc tgtttgcatc gtcctttctg
     4981 atgatacttg ttctgatgag aagattcgga tgaatagagt tgttcggaat aaccttcgtg
     5041 tacgcctagg ggatgtcatc agcatccagc catgccctga tgtgaagtac ggcaaacgta
     5101 tccatgtgct gcccattgat gacacagtgg aaggcattac tggtaatctc ttcgaggtat
     5161 accttaagcc gtacttcctg gaagcgtatc gacccatccg gaaaggagac atttttcttg
     5221 tccgtggtgg gatgcgtgct gtggagttca aagtggtgga aacagatcct agcccttatt
     5281 gcattgttgc tccagacaca gtgatccact gcgaagggga gcctatcaaa cgagaggatg
     5341 aggaagagtc cttgaatgaa gtagggtatg atgacattgg tggctgcagg aagcagctag
     5401 ctcagataaa ggagatggtg gaactgcccc tgagacatcc tgccctcttt aaggcaattg
     5461 gtgtgaagcc tcctagagga atcctgcttt acggacctcc tggaacagga aagaccctga
     5521 ttgctcgagc tgtagcaaat gagactggag ccttcttctt cttgatcaat ggtcctgaga
     5581 tcatgagcaa attggctggt gagtctgaga gcaaccttcg taaagccttt gaggaggctg
     5641 agaagaatgc tcctgccatc atcttcattg atgagctaga tgccatcgct cccaaaagag
     5701 agaaaactca tggcgaggtg gagcggcgca ttgtatcaca gttgttgacc ctcatggatg
     5761 gcctaaagca gagggcacat gtgattgtta tggcagcaac caacagaccc aacagcattg
     5821 acccagctct acggcgattt ggtcgctttg acagggaggt agatattgga attcctgatg
     5881 ctacaggacg cttagagatt cttcagatcc ataccaagaa catgaagctg gcagatgatg
     5941 tggacctgga acaggtagcc aatgagactc acgggcatgt gggtgctgac ttagcagccc
     6001 tgtgctcaga ggctgctctg caagccatcc gcaagaagat ggatctcatt gacctagagg
     6061 atgagaccat tgatgccgag gtcatgaact ctctagcagt tactatggat gacttccggt
     6121 gggccttgag ccagagtaac ccatcagcac tgcgggaaac cgtggtagag gtgccacagg
     6181 taacctggga agacatcggg ggcctagagg atgtcaaacg tgagctacag gagctggtcc
     6241 agtatcctgt ggagcaccca gacaaattcc tgaagtttgg catgacacct tccaagggag
     6301 ttctgttcta tggacctcct ggctgtggga aaactttgtt ggccaaagcc attgctaatg
     6361 aatgccaggc caacttcatc tccatcaagg gtcctgagct gctcaccatg tggtttgggg
     6421 agtctgaggc caatgtcaga gaaatctttg acaaggcccg ccaagctgcc ccctgtgtgc
     6481 tattctttga tgagctggat tcgattgcca aggctcgtgg aggtaacatt ggagatggtg
     6541 gtggggctgc tgaccgagtc atcaaccaga tcctgacaga aatggatggc atgtccacta
     6601 aaaaaaatgt gttcatcatt ggcgctacca accggcctga catcattgat cctgccatcc
     6661 tcagacctgg ccgtcttgat cagctcatct acatcccact tcctgatgag aagtcccgtg
     6721 ttgccatcct caaggctaac ctgcgcaagt ccccagttgc caaggatgtg gacttggagt
     6781 tcctggctaa aatgactaat ggcttctctg gagctgacct gacagagatt tgccagcgtg
     6841 cttgcaagct ggccatccgt gaatccatcg agagtgagat taggcgagaa cgagagaggc
     6901 agacaaaccc atcagccatg gaggtagaag aggatgatcc agtgcctgag atccgtcgag
     6961 atcactttga agaagccatg cgctttgcgc gccgttctgt cagtgacaat gacattcgga
     7021 agtatgagat gtttgcccag acccttcagc agagtcgggg ctttggcagc ttcagattcc
     7081 cttcagggaa ccagggtgga gctggcccca gtcagggcag tggaggcggc acaggtggca
     7141 gtgtatacac agaagacaat gatgatgacc tgtatggcta agagctcact agtcgcggcc
     7201 gctttcgaat ctagagcctg cagtctcgac aagcttgtcg agaagtacta gaggatcata
     7261 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     7321 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     7381 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     7441 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgatca
     7501 ctgcttgagc ctagaagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     7561 tgccaccgct gagcaataac tatcataacc cctaggagat ccgaaccaga taagtgaaat
     7621 ctagttccaa actattttgt catttttaat tttcgtatta gcttacgacg ctacacccag
     7681 ttcccatcta ttttgtcact cttccctaaa taatccttaa aaactccatt tccacccctc
     7741 ccagttccca actattttgt ccgcccacag cggggcattt ttcttcctgt tatgttttta
     7801 atcaaacatc ctgccaactc catgtgacaa accgtcatct tcggctactt t
//
