LOCUS       sequence-218698-        5892 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of His-I3 V41A / W43 A in insect cells (Baculovirus 
            system).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5892)
  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 5892)
  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-I3 V41A / W43A
FEATURES             Location/Qualifiers
     source          1..5892
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        33..124
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     49..66
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             185..202
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             224..244
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    825..959
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(862..881)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     916..935
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1069..1234
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      1418..1872
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1505..1524)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1714..1735
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1898..2002
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2003..2071,2072..2863)
                     /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(2221..2240)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3034..3622
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3523..3542
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3776..3793
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  3927..4151
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4218..4751)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4940..4945,4946..4962,4963..4968))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(5364..5397)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    5587..5635
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5660..5679
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(5771..5880)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
ORIGIN
        1 tcgattcgcg acctactccg gaatattaat agatcatgga gataattaaa atgataacca
       61 tctcgcaaat aaataagtat tttactgttt tcgtaacagt tttgtaataa aaaaacctat
      121 aaatattccg gattattcat accgtcccac catcgggcgc ggatctacaa ccatgtcgta
      181 ctaccatcac catcaccatc acgattacga tatcccaacg accgaaaacc tgtattttca
      241 gggcgccatg ggatccatgg ccgaggcagg ggctgggctg agcgagaccg tcactgagac
      301 aacggttacc gtgacaaccg agcccgagaa ccggagcctt accatcaaac ttcggaaacg
      361 gaagccagag aaaaaggcag aagcgacaag tgacactgtg gacaatgaac acatgggccg
      421 ccgctcatcc aaatgctgct gtatttatga gaaacctcgg gcctttggcg agagctccac
      481 ggaaagtgat gaggaggaag aagagggctg tggtcataca cactgtgtac gtggccaccg
      541 caaaggacgg cgtcgtgcaa ccctaggacc gacccccacc acccctcccc agcctcctga
      601 cccttcccag ccccctccag ggccaatgca gcactaatga gtcgacgagc tcactagtcg
      661 cggccgcttt cgaatctaga gcctgcagtc tcgacaagct tgtcgagaag tactagagga
      721 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
      781 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
      841 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
      901 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct
      961 gatcactgct tgagcctaga agatccggct gctaacaaag cccgaaagga agctgagttg
     1021 gctgctgcca ccgctgagca ataactatca taacccctag gagatccgaa ccagataagt
     1081 gaaatctagt tccaaactat tttgtcattt ttaattttcg tattagctta cgacgctaca
     1141 cccagttccc atctattttg tcactcttcc ctaaataatc cttaaaaact ccatttccac
     1201 ccctcccagt tcccaactat tttgtccgcc cacagcgggg catttttctt cctgttatgt
     1261 ttttaatcaa acatcctgcc aactccatgt gacaaaccgt catcttcggc tactttttct
     1321 ctgtcacaga atgaaaattt ttctgtcatc tcttcgttat taatgtttgt aattgactga
     1381 atatcaacgc ttatttgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     1441 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     1501 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     1561 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     1621 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     1681 cgccctttga cgttggagtc cacgttctta atagtggact cttgttccaa actggaacaa
     1741 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     1801 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     1861 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     1921 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     1981 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2041 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2101 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2161 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     2221 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     2281 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     2341 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     2401 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     2461 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     2521 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     2581 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     2641 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2701 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     2761 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     2821 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     2881 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     2941 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3001 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3061 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3121 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3181 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3241 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3301 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3361 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3421 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3481 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3541 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3601 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3661 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3721 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3781 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     3841 cggtatttca caccgcatag accagccgcg taacctggca aaatcggtta cggttgagta
     3901 ataaatggat gccctgcgta agcgggtgtg ggcggacaat aaagtcttaa actgaacaaa
     3961 atagatctaa actatgacaa taaagtctta aactagacag aatagttgta aactgaaatc
     4021 agtccagtta tgctgtgaaa aagcatactg gacttttgtt atggctaaag caaactcttc
     4081 attttctgaa gtgcaaattg cccgtcgtat taaagagggg cgtggccaag ggcatggtaa
     4141 agactatatt cgcggcgttg tgacaattta ccgaacaact ccgcggccgg gaagccgatc
     4201 tcggcttgaa cgaattgtta ggtggcggta cttgggtcga tatcaaagtg catcacttct
     4261 tcccgtatgc ccaactttgt atagagagcc actgcgggat cgtcaccgta atctgcttgc
     4321 acgtagatca cataagcacc aagcgcgttg gcctcatgct tgaggagatt gatgagcgcg
     4381 gtggcaatgc cctgcctccg gtgctcgccg gagactgcga gatcatagat atagatctca
     4441 ctacgcggct gctcaaactt gggcagaacg taagccgcga gagcgccaac aaccgcttct
     4501 tggtcgaagg cagcaagcgc gatgaatgtc ttactacgga gcaagttccc gaggtaatcg
     4561 gagtccggct gatgttggga gtaggtggct acgtctccga actcacgacc gaaaagatca
     4621 agagcagccc gcatggattt gacttggtca gggccgagcc tacatgtgcg aatgatgccc
     4681 atacttgagc cacctaactt tgttttaggg cgactgccct gctgcgtaac atcgttgctg
     4741 ctgcgtaaca tcgttgctgc tccataacat caaacatcga cccacggcgt aacgcgcttg
     4801 ctgcttggat gcccgaggca tagactgtac aaaaaaacag tcataacaag ccatgaaaac
     4861 cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg
     4921 catacgctac ttgcattaca gtttacgaac cgaacaggct tatgtcaact gggtttgtgc
     4981 cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg ggcagcagcg aagtcgaggc
     5041 atttctgtcc tggctggcga acgagcgcaa ggtttcggtc tccacgcatc gtcaggcatt
     5101 ggcggccttg ctgttcttct acggcaaggt gctgtgcacg gatctgccct tgcttcagga
     5161 gatcggtaga cctcggccgt cgcggcgctt gccggtggtg ctgaccccgg atgaagtggt
     5221 tcgcatcctc ggttttctgg aaggcgagca tcgtttgttc gcccaggact ctagctatag
     5281 ttctagtggt tggctacagc tttgtttgta ctatcaacag gttgaactgc tgatcaacag
     5341 atcctctacg cggccgcggt accataactt cgtatagcat acattatacg aagttatctg
     5401 gtttaaacgt acccgtagtg gctatggcag ggcttgccgc cccgacgttg gctgcgagcc
     5461 ctgggccttc acccgaactt gggggttggg gtggggaaaa ggaagaaacg cgggcgtatt
     5521 ggtcccaatg gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc
     5581 gtttatgaac aaacgaccca acacccgtgc gttttattct gtctttttat tgccgtcata
     5641 gcgcgggttc cttccggtat tgtctccttc cgtgtttcag ttagcctccc ccatctcccg
     5701 gtaccgcatg ctatgcatca gctgctagca ccatggctcg agatcccggg tgatcaagtc
     5761 ttcgtcgagt gattgtaaat aaaatgtaat ttacagtata gtattttaat taatatacaa
     5821 atgatttgat aataattctt atttaactat aatatattgt gttgggttga attaaaggtc
     5881 cgtatactag ta
//
