LOCUS       sequence-218665-        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"
     promoter        complement(join(413..418,419..435,436..441))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(837..870)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1060..1108
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1133..1152
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(2225..2334)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        2379..2470
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2395..2412
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2531..2548
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2570..2590
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     regulatory      2601..2610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3896..4030
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3933..3952)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3987..4006
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4140..4305
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4489..4943
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4576..4595)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4785..4806
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4969..5073
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5074..5142,5143..5934)
                     /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(5292..5311)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6105..6693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6594..6613
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6847..6864
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6998..7222
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(join(7289..7598,1..224))
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 gtcttactac ggagcaagtt cccgaggtaa tcggagtccg gctgatgttg ggagtaggtg
       61 gctacgtctc cgaactcacg accgaaaaga tcaagagcag cccgcatgga tttgacttgg
      121 tcagggccga gcctacatgt gcgaatgatg cccatacttg agccacctaa ctttgtttta
      181 gggcgactgc cctgctgcgt aacatcgttg ctgctgcgta acatcgttgc tgctccataa
      241 catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag gcatagactg
      301 tacaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac caccgctgcg
      361 ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt acagtttacg
      421 aaccgaacag gcttatgtca actgggtttg tgccttcatc cgtttccacg gtgtgcgtca
      481 cccggcaacc ttgggcagca gcgaagtcga ggcatttctg tcctggctgg cgaacgagcg
      541 caaggtttcg gtctccacgc atcgtcaggc attggcggcc ttgctgttct tctacggcaa
      601 ggtgctgtgc acggatctgc ccttgcttca ggagatcggt agacctcggc cgtcgcggcg
      661 cttgccggtg gtgctgaccc cggatgaagt ggttcgcatc ctcggttttc tggaaggcga
      721 gcatcgtttg ttcgcccagg actctagcta tagttctagt ggttggctac agctttgttt
      781 gtactatcaa caggttgaac tgctgatcaa cagatcctct acgcggccgc ggtaccataa
      841 cttcgtatag catacattat acgaagttat ctggtttaaa cgtacccgta gtggctatgg
      901 cagggcttgc cgccccgacg ttggctgcga gccctgggcc ttcacccgaa cttgggggtt
      961 ggggtgggga aaaggaagaa acgcgggcgt attggtccca atggggtctc ggtggggtat
     1021 cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac ccaacacccg
     1081 tgcgttttat tctgtctttt tattgccgtc atagcgcggg ttccttccgg tattgtctcc
     1141 ttccgtgttt cagttagcct cccccatctc ccggtaccgc atgctatgca tcagctgcta
     1201 gcaccatggc tcgagttatc atttctttgc ttgctttgtg atcatacccc ttggaggcgt
     1261 tacaggtctc gtggcatttg gcttcttttt ctctgcaggc tttaaaatct gaaaagaaca
     1321 cattagtgtt tcatccacac tcatcatggc acctgcattg tcaaactctc cgcaataatt
     1381 gggcgcagaa aacagagtga ccaactgcct ctttgcaaaa aattcatatc catcttcaac
     1441 cacctgatgg gctctacata taagatccaa atcatgctta tggagaaatt ttgcaaccac
     1501 ttctgcacca aatgtgaagg acactcctct gtcattttca ccccagccta agacatcttt
     1561 atcggggtca gaccacaaaa gatcacaaag aagaccttga tctggtacat cagttggtcg
     1621 cataattcgc cgaatctgct ccatagattg aagatctggt gataaacctc catgacagca
     1681 gaatatcttc tcatccacga tggctgctat cggtaaacag ttaaaacagt ctgtgaaagt
     1741 tttccatagt ttaatgttgt atcttctttt acattcatca taaaatccat aaattctgtt
     1801 gatgctggca cattcatggt tccctctgag aagaaaaaaa ttctcaggat attttatttt
     1861 gtaggccagt aagaggcaga tcgtctccaa tgactgcttt cccctgtcca catagtcccc
     1921 aagaaacagg tagttgcttt ctggtgggaa accaccgtac tcaaaaagtc gcagcaaatc
     1981 atagtattgt ccatggatgt caccacatat tttgagtggt gcttcaagtt ctagtaggat
     2041 aggctgactg agaaagattt cacgagactt taagcacagt cctctgattt cattctcctg
     2101 aagctggaca ttcttaccag gcttggaccc tctcacttcc agcagccgtt ggataatgct
     2161 gtcgatgttg agtttatcta aatccgccat ctcgagatcc cgggtgatca agtcttcgtc
     2221 gagtgattgt aaataaaatg taatttacag tatagtattt taattaatat acaaatgatt
     2281 tgataataat tcttatttaa ctataatata ttgtgttggg ttgaattaaa ggtccgtata
     2341 ctagtatcga ttcgcgacct actccggaat attaatagat catggagata attaaaatga
     2401 taaccatctc gcaaataaat aagtatttta ctgttttcgt aacagttttg taataaaaaa
     2461 acctataaat attccggatt attcataccg tcccaccatc gggcgcggat ctacaaccat
     2521 gtcgtactac catcaccatc accatcacga ttacgatatc ccaacgaccg aaaacctgta
     2581 ttttcagggc gccatgggat ccatggcggc ggaacgcggc gcggggcagc aacagtcgca
     2641 ggagatgatg gaggttgaca ggcgggtcga gtctgaagaa tccggcgatg aagaagggaa
     2701 gaaacacagc agtggcatcg tggccgacct cagtgaacag agcctgaagg atggggagga
     2761 gcggggggag gaggacccag aagaagaaca tgagctgcct gtggacatgg aaaccatcaa
     2821 cctggacaga gatgcagagg atgttgattt gaatcactat cgcataggga agattgaagg
     2881 atttgaggta ctgaagaaag tgaagactct ctgcctccgc caaaatttaa ttaaatgcat
     2941 tgagaatctg gaggagctac agagtcttcg agagctggat ctttacgaca accagatcaa
     3001 gaagattgag aatctggagg cgctaacaga gctggagatt ctagatattt cttttaatct
     3061 gctgagaaac atcgaagggg ttgacaagtt gacacgactg aaaaaactct tcttggtcaa
     3121 caataaaatc agtaaaattg agaacttaag caacttacat caactacaga tgctagagct
     3181 gggatctaac cgcatccggg caatcgaaaa tatcgacacc ttaaccaacc tggagagttt
     3241 gtttttgggg aaaaacaaaa ttactaaact tcagaacctg gatgcgctca ccaacctgac
     3301 agtcctcagt atgcagagca accggctgac caagatcgag ggtctgcaga acctggtgaa
     3361 cctgcgggag ctgtacctta gccacaatgg catcgaggtc atcgagggcc tggagaacaa
     3421 taacaaactc acgatgttgg acattgcatc aaatagaatc aaaaagattg aaaatatcag
     3481 ccatctaaca gagctgcaag agttctggat gaacgacaat ctccttgaga gctggagcga
     3541 cctcgacgag ctgaagggag ccaggagcct ggagacagtg tacctggagc ggaacccctt
     3601 gcagaaggac ccccagtacc ggcggaaggt catgctcgcc ctcccctccg tgcggcagat
     3661 cgatgccacg ttcgtcaggt tctgataagg atccggaatt caaaggccta cgtcgacgag
     3721 ctcactagtc gcggccgctt tcgaatctag agcctgcagt ctcgacaagc ttgtcgagaa
     3781 gtactagagg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3841 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3901 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3961 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4021 tgtctggatc tgatcactgc ttgagcctag aagatccggc tgctaacaaa gcccgaaagg
     4081 aagctgagtt ggctgctgcc accgctgagc aataactatc ataaccccta ggagatccga
     4141 accagataag tgaaatctag ttccaaacta ttttgtcatt tttaattttc gtattagctt
     4201 acgacgctac acccagttcc catctatttt gtcactcttc cctaaataat ccttaaaaac
     4261 tccatttcca cccctcccag ttcccaacta ttttgtccgc ccacagcggg gcatttttct
     4321 tcctgttatg tttttaatca aacatcctgc caactccatg tgacaaaccg tcatcttcgg
     4381 ctactttttc tctgtcacag aatgaaaatt tttctgtcat ctcttcgtta ttaatgtttg
     4441 taattgactg aatatcaacg cttatttgca gcctgaatgg cgaatgggac gcgccctgta
     4501 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4561 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4621 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4681 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4741 agacggtttt tcgccctttg acgttggagt ccacgttctt aatagtggac tcttgttcca
     4801 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4861 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     4921 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     4981 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5041 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5101 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5161 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5221 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5281 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5341 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5401 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5461 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5521 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5581 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5641 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5701 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5761 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5821 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5881 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5941 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6001 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6061 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6121 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6181 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6241 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6301 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6361 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6421 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6481 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6541 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6601 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6661 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6721 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6781 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6841 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     6901 acgcatctgt gcggtatttc acaccgcata gaccagccgc gtaacctggc aaaatcggtt
     6961 acggttgagt aataaatgga tgccctgcgt aagcgggtgt gggcggacaa taaagtctta
     7021 aactgaacaa aatagatcta aactatgaca ataaagtctt aaactagaca gaatagttgt
     7081 aaactgaaat cagtccagtt atgctgtgaa aaagcatact ggacttttgt tatggctaaa
     7141 gcaaactctt cattttctga agtgcaaatt gcccgtcgta ttaaagaggg gcgtggccaa
     7201 gggcatggta aagactatat tcgcggcgtt gtgacaattt accgaacaac tccgcggccg
     7261 ggaagccgat ctcggcttga acgaattgtt aggtggcggt acttgggtcg atatcaaagt
     7321 gcatcacttc ttcccgtatg cccaactttg tatagagagc cactgcggga tcgtcaccgt
     7381 aatctgcttg cacgtagatc acataagcac caagcgcgtt ggcctcatgc ttgaggagat
     7441 tgatgagcgc ggtggcaatg ccctgcctcc ggtgctcgcc ggagactgcg agatcataga
     7501 tatagatctc actacgcggc tgctcaaact tgggcagaac gtaagccgcg agagcgccaa
     7561 caaccgcttc ttggtcgaag gcagcaagcg cgatgaat
//
