LOCUS       sequence-214245-        7598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of His-SDS22 / PP1gamma dimer in insect cells 
            (Baculovirus system).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7598)
  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 7598)
  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 / PP1gamma
FEATURES             Location/Qualifiers
     source          1..7598
                     /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(5436..5545)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        5590..5681
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     5606..5623
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             5742..5759
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5781..5801
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     regulatory      5812..5821
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    7107..7241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7144..7163)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7198..7217
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  7351..7516
                     /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 ctcgagttat catttctttg
     4441 cttgctttgt gatcataccc cttggaggcg ttacaggtct cgtggcattt ggcttctttt
     4501 tctctgcagg ctttaaaatc tgaaaagaac acattagtgt ttcatccaca ctcatcatgg
     4561 cacctgcatt gtcaaactct ccgcaataat tgggcgcaga aaacagagtg accaactgcc
     4621 tctttgcaaa aaattcatat ccatcttcaa ccacctgatg ggctctacat ataagatcca
     4681 aatcatgctt atggagaaat tttgcaacca cttctgcacc aaatgtgaag gacactcctc
     4741 tgtcattttc accccagcct aagacatctt tatcggggtc agaccacaaa agatcacaaa
     4801 gaagaccttg atctggtaca tcagttggtc gcataattcg ccgaatctgc tccatagatt
     4861 gaagatctgg tgataaacct ccatgacagc agaatatctt ctcatccacg atggctgcta
     4921 tcggtaaaca gttaaaacag tctgtgaaag ttttccatag tttaatgttg tatcttcttt
     4981 tacattcatc ataaaatcca taaattctgt tgatgctggc acattcatgg ttccctctga
     5041 gaagaaaaaa attctcagga tattttattt tgtaggccag taagaggcag atcgtctcca
     5101 atgactgctt tcccctgtcc acatagtccc caagaaacag gtagttgctt tctggtggga
     5161 aaccaccgta ctcaaaaagt cgcagcaaat catagtattg tccatggatg tcaccacata
     5221 ttttgagtgg tgcttcaagt tctagtagga taggctgact gagaaagatt tcacgagact
     5281 ttaagcacag tcctctgatt tcattctcct gaagctggac attcttacca ggcttggacc
     5341 ctctcacttc cagcagccgt tggataatgc tgtcgatgtt gagtttatct aaatccgcca
     5401 tctcgagatc ccgggtgatc aagtcttcgt cgagtgattg taaataaaat gtaatttaca
     5461 gtatagtatt ttaattaata tacaaatgat ttgataataa ttcttattta actataatat
     5521 attgtgttgg gttgaattaa aggtccgtat actagtatcg attcgcgacc tactccggaa
     5581 tattaataga tcatggagat aattaaaatg ataaccatct cgcaaataaa taagtatttt
     5641 actgttttcg taacagtttt gtaataaaaa aacctataaa tattccggat tattcatacc
     5701 gtcccaccat cgggcgcgga tctacaacca tgtcgtacta ccatcaccat caccatcacg
     5761 attacgatat cccaacgacc gaaaacctgt attttcaggg cgccatggga tccatggcgg
     5821 cggaacgcgg cgcggggcag caacagtcgc aggagatgat ggaggttgac aggcgggtcg
     5881 agtctgaaga atccggcgat gaagaaggga agaaacacag cagtggcatc gtggccgacc
     5941 tcagtgaaca gagcctgaag gatggggagg agcgggggga ggaggaccca gaagaagaac
     6001 atgagctgcc tgtggacatg gaaaccatca acctggacag agatgcagag gatgttgatt
     6061 tgaatcacta tcgcataggg aagattgaag gatttgaggt actgaagaaa gtgaagactc
     6121 tctgcctccg ccaaaattta attaaatgca ttgagaatct ggaggagcta cagagtcttc
     6181 gagagctgga tctttacgac aaccagatca agaagattga gaatctggag gcgctaacag
     6241 agctggagat tctagatatt tcttttaatc tgctgagaaa catcgaaggg gttgacaagt
     6301 tgacacgact gaaaaaactc ttcttggtca acaataaaat cagtaaaatt gagaacttaa
     6361 gcaacttaca tcaactacag atgctagagc tgggatctaa ccgcatccgg gcaatcgaaa
     6421 atatcgacac cttaaccaac ctggagagtt tgtttttggg gaaaaacaaa attactaaac
     6481 ttcagaacct ggatgcgctc accaacctga cagtcctcag tatgcagagc aaccggctga
     6541 ccaagatcga gggtctgcag aacctggtga acctgcggga gctgtacctt agccacaatg
     6601 gcatcgaggt catcgagggc ctggagaaca ataacaaact cacgatgttg gacattgcat
     6661 caaatagaat caaaaagatt gaaaatatca gccatctaac agagctgcaa gagttctgga
     6721 tgaacgacaa tctccttgag agctggagcg acctcgacga gctgaaggga gccaggagcc
     6781 tggagacagt gtacctggag cggaacccct tgcagaagga cccccagtac cggcggaagg
     6841 tcatgctcgc cctcccctcc gtgcggcaga tcgatgccac gttcgtcagg ttctgataag
     6901 gatccggaat tcaaaggcct acgtcgacga gctcactagt cgcggccgct ttcgaatcta
     6961 gagcctgcag tctcgacaag cttgtcgaga agtactagag gatcataatc agccatacca
     7021 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     7081 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     7141 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     7201 gtttgtccaa actcatcaat gtatcttatc atgtctggat ctgatcactg cttgagccta
     7261 gaagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     7321 caataactat cataacccct aggagatccg aaccagataa gtgaaatcta gttccaaact
     7381 attttgtcat ttttaatttt cgtattagct tacgacgcta cacccagttc ccatctattt
     7441 tgtcactctt ccctaaataa tccttaaaaa ctccatttcc acccctccca gttcccaact
     7501 attttgtccg cccacagcgg ggcatttttc ttcctgttat gtttttaatc aaacatcctg
     7561 ccaactccat gtgacaaacc gtcatcttcg gctacttt
//
