LOCUS       sequence-218695-        6617 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of His-SDS22 in insect cells (Baculovirus system).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6617)
  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 6617)
  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-SDS22
FEATURES             Location/Qualifiers
     source          1..6617
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..121
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     46..63
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             182..199
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             221..241
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     regulatory      252..261
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1547..1681
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1584..1603)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1638..1657
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1791..1956
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2140..2594
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2227..2246)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2436..2457
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2620..2724
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2725..2793,2794..3585)
                     /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(2943..2962)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3756..4344
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4245..4264
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4498..4515
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4649..4873
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4940..5473)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5662..5667,5668..5684,5685..5690))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(6086..6119)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    6309..6357
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     6382..6401
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(6493..6602)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
ORIGIN
        1 attcgcgacc tactccggaa tattaataga tcatggagat aattaaaatg ataaccatct
       61 cgcaaataaa taagtatttt actgttttcg taacagtttt gtaataaaaa aacctataaa
      121 tattccggat tattcatacc gtcccaccat cgggcgcgga tctacaacca tgtcgtacta
      181 ccatcaccat caccatcacg attacgatat cccaacgacc gaaaacctgt attttcaggg
      241 cgccatggga tccatggcgg cggaacgcgg cgcggggcag caacagtcgc aggagatgat
      301 ggaggttgac aggcgggtcg agtctgaaga atccggcgat gaagaaggga agaaacacag
      361 cagtggcatc gtggccgacc tcagtgaaca gagcctgaag gatggggagg agcgggggga
      421 ggaggaccca gaagaagaac atgagctgcc tgtggacatg gaaaccatca acctggacag
      481 agatgcagag gatgttgatt tgaatcacta tcgcataggg aagattgaag gatttgaggt
      541 actgaagaaa gtgaagactc tctgcctccg ccaaaattta attaaatgca ttgagaatct
      601 ggaggagcta cagagtcttc gagagctgga tctttacgac aaccagatca agaagattga
      661 gaatctggag gcgctaacag agctggagat tctagatatt tcttttaatc tgctgagaaa
      721 catcgaaggg gttgacaagt tgacacgact gaaaaaactc ttcttggtca acaataaaat
      781 cagtaaaatt gagaacttaa gcaacttaca tcaactacag atgctagagc tgggatctaa
      841 ccgcatccgg gcaatcgaaa atatcgacac cttaaccaac ctggagagtt tgtttttggg
      901 gaaaaacaaa attactaaac ttcagaacct ggatgcgctc accaacctga cagtcctcag
      961 tatgcagagc aaccggctga ccaagatcga gggtctgcag aacctggtga acctgcggga
     1021 gctgtacctt agccacaatg gcatcgaggt catcgagggc ctggagaaca ataacaaact
     1081 cacgatgttg gacattgcat caaatagaat caaaaagatt gaaaatatca gccatctaac
     1141 agagctgcaa gagttctgga tgaacgacaa tctccttgag agctggagcg acctcgacga
     1201 gctgaaggga gccaggagcc tggagacagt gtacctggag cggaacccct tgcagaagga
     1261 cccccagtac cggcggaagg tcatgctcgc cctcccctcc gtgcggcaga tcgatgccac
     1321 gttcgtcagg ttctgataag gatccggaat tcaaaggcct acgtcgacga gctcactagt
     1381 cgcggccgct ttcgaatcta gagcctgcag tctcgacaag cttgtcgaga agtactagag
     1441 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     1501 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     1561 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     1621 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     1681 ctgatcactg cttgagccta gaagatccgg ctgctaacaa agcccgaaag gaagctgagt
     1741 tggctgctgc caccgctgag caataactat cataacccct aggagatccg aaccagataa
     1801 gtgaaatcta gttccaaact attttgtcat ttttaatttt cgtattagct tacgacgcta
     1861 cacccagttc ccatctattt tgtcactctt ccctaaataa tccttaaaaa ctccatttcc
     1921 acccctccca gttcccaact attttgtccg cccacagcgg ggcatttttc ttcctgttat
     1981 gtttttaatc aaacatcctg ccaactccat gtgacaaacc gtcatcttcg gctacttttt
     2041 ctctgtcaca gaatgaaaat ttttctgtca tctcttcgtt attaatgttt gtaattgact
     2101 gaatatcaac gcttatttgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     2161 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2221 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2281 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2341 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2401 ttcgcccttt gacgttggag tccacgttct taatagtgga ctcttgttcc aaactggaac
     2461 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     2521 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     2581 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2641 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     2701 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     2761 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     2821 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     2881 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     2941 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     3001 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3061 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3121 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3181 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3241 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     3301 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     3361 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     3421 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     3481 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     3541 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     3601 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3661 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3721 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3781 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3841 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3901 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3961 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4021 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4081 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4141 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4201 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4261 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4321 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4381 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4441 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     4501 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     4561 tgcggtattt cacaccgcat agaccagccg cgtaacctgg caaaatcggt tacggttgag
     4621 taataaatgg atgccctgcg taagcgggtg tgggcggaca ataaagtctt aaactgaaca
     4681 aaatagatct aaactatgac aataaagtct taaactagac agaatagttg taaactgaaa
     4741 tcagtccagt tatgctgtga aaaagcatac tggacttttg ttatggctaa agcaaactct
     4801 tcattttctg aagtgcaaat tgcccgtcgt attaaagagg ggcgtggcca agggcatggt
     4861 aaagactata ttcgcggcgt tgtgacaatt taccgaacaa ctccgcggcc gggaagccga
     4921 tctcggcttg aacgaattgt taggtggcgg tacttgggtc gatatcaaag tgcatcactt
     4981 cttcccgtat gcccaacttt gtatagagag ccactgcggg atcgtcaccg taatctgctt
     5041 gcacgtagat cacataagca ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg
     5101 cggtggcaat gccctgcctc cggtgctcgc cggagactgc gagatcatag atatagatct
     5161 cactacgcgg ctgctcaaac ttgggcagaa cgtaagccgc gagagcgcca acaaccgctt
     5221 cttggtcgaa ggcagcaagc gcgatgaatg tcttactacg gagcaagttc ccgaggtaat
     5281 cggagtccgg ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat
     5341 caagagcagc ccgcatggat ttgacttggt cagggccgag cctacatgtg cgaatgatgc
     5401 ccatacttga gccacctaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc
     5461 tgctgcgtaa catcgttgct gctccataac atcaaacatc gacccacggc gtaacgcgct
     5521 tgctgcttgg atgcccgagg catagactgt acaaaaaaac agtcataaca agccatgaaa
     5581 accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag
     5641 cgcatacgct acttgcatta cagtttacga accgaacagg cttatgtcaa ctgggtttgt
     5701 gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct tgggcagcag cgaagtcgag
     5761 gcatttctgt cctggctggc gaacgagcgc aaggtttcgg tctccacgca tcgtcaggca
     5821 ttggcggcct tgctgttctt ctacggcaag gtgctgtgca cggatctgcc cttgcttcag
     5881 gagatcggta gacctcggcc gtcgcggcgc ttgccggtgg tgctgacccc ggatgaagtg
     5941 gttcgcatcc tcggttttct ggaaggcgag catcgtttgt tcgcccagga ctctagctat
     6001 agttctagtg gttggctaca gctttgtttg tactatcaac aggttgaact gctgatcaac
     6061 agatcctcta cgcggccgcg gtaccataac ttcgtatagc atacattata cgaagttatc
     6121 tggtttaaac gtacccgtag tggctatggc agggcttgcc gccccgacgt tggctgcgag
     6181 ccctgggcct tcacccgaac ttgggggttg gggtggggaa aaggaagaaa cgcgggcgta
     6241 ttggtcccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc
     6301 gcgtttatga acaaacgacc caacacccgt gcgttttatt ctgtcttttt attgccgtca
     6361 tagcgcgggt tccttccggt attgtctcct tccgtgtttc agttagcctc ccccatctcc
     6421 cggtaccgca tgctatgcat cagctgctag caccatggct cgagatcccg ggtgatcaag
     6481 tcttcgtcga gtgattgtaa ataaaatgta atttacagta tagtatttta attaatatac
     6541 aaatgatttg ataataattc ttatttaact ataatatatt gtgttgggtt gaattaaagg
     6601 tccgtatact agtatcg
//
