LOCUS       sequence-376490-        6466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PALI1 from insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6466)
  AUTHORS   Zhang Q, Agius SC, Flanigan SF, Uckelmann M, Levina V, Owen BM, 
            Davidovich C
  TITLE     PALI1 facilitates DNA and nucleosome binding by PRC2 and triggers an
            allosteric activation of catalysis.
  JOURNAL   Nat Commun. 2021 Jul 28;12(1):4592. doi: 10.1038/s41467-021-24866-3.
  PUBMED    34321472
REFERENCE   2  (bases 1 to 6466)
  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: "Nat 
            Commun."; date: "2021-07-28"; volume: "12(1):4592. doi"; pages: " 
            10.1038/s41467-021-24866-3"
COMMENT     Sequence Label: PALI1_PIR_pFB1
FEATURES             Location/Qualifiers
     source          1..6466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        186..277
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     202..219
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             338..355
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             362..1459
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1316..1325
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1433..1456
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1496..1519
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    2270..2404
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2307..2326)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2361..2380
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2433..2598
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2782..3236
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2869..2888)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3078..3099
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3262..3366
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3367..3435,3436..4227)
                     /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(3585..3604)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4398..4986
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4887..4906
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5129..5146
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  5280..5504
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(5571..6104)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(6274..6279,6280..6296,6297..6302))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 tgtgcacgga tctgccctgg cttcaggaga tcggaagacc tcggccgtcg cggcgcttgc
       61 cggtggtgct gaccccggat gaagtggttc gcatcctcgg ttttctggaa ggcgagcatc
      121 gtttgttcgc ccaggactct agctatagtt ctagtggttg gctacatact ccggaatatt
      181 aatagatcat ggagataatt aaaatgataa ccatctcgca aataaataag tattttactg
      241 ttttcgtaac agttttgtaa taaaaaaacc tataaatatt ccggattatt cataccgtcc
      301 caccatcggg cgcggatctc ggtccgaaac catgggtcat caccatcatc atcacggtac
      361 caaaactgaa gaaggtaaac tggtaatctg gattaacggc gataaaggct ataacggtct
      421 cgctgaagtc ggtaagaaat tcgagaaaga taccggaatt aaagtcaccg ttgagcatcc
      481 ggataaactg gaagagaaat tcccacaggt tgcggcaact ggcgatggcc ctgacattat
      541 cttctgggca cacgaccgct ttggtggcta cgctcaatct ggcctgttgg ctgaaatcac
      601 cccggacaaa gcgttccagg acaagctgta tccgtttacc tgggatgccg tacgttacaa
      661 cggcaagctg attgcttacc cgatcgctgt tgaagcgtta tcgctgattt ataacaaaga
      721 tctgctgccg aacccgccaa aaacctggga agagatcccg gcgctggata aagaactgaa
      781 agcgaaaggt aagagcgcgc tgatgttcaa cctgcaagaa ccgtacttca cctggccgct
      841 gattgctgct gacgggggtt atgcgttcaa gtatgaaaac ggcaagtacg acattaaaga
      901 cgtgggcgtg gataacgctg gcgcgaaagc gggtctgacc ttcctggttg acctgattaa
      961 aaacaaacac atgaatgcag acaccgatta ctccatcgca gaagctgcct ttaataaagg
     1021 cgaaacagcg atgaccatca acggcccgtg ggcatggtcc aacatcgaca ccagcaaagt
     1081 gaattatggt gtaacggtac tgccgacctt caagggtcaa ccatccaaac cgttcgttgg
     1141 cgtgctgagc gcaggtatta acgccgccag tccgaacaaa gagctggcaa aagagttcct
     1201 cgaaaactat ctgctgactg atgaaggtct ggaagcggtt aataaagaca aaccgctggg
     1261 tgccgtagcg ctgaagtctt acgaggaaga gttggcgaaa gatccacgta ttgccgccac
     1321 catggaaaac gcccagaaag gtgaaatcat gccgaacatc ccgcagatgt ccgctttctg
     1381 gtatgccgtg cgtactgcgg tgatcaacgc cgccagcggt cgtcagactg tcgatgaagc
     1441 cctgaaagac gcgcagactg cggccgcagg caccgattac gatatcccaa cgaccctgga
     1501 agttctgttc caggggcccg ggacttcaga aaaggaagct gcacaagtaa accccataat
     1561 gccaaaggaa aatggagctt cagagagtgg agacccccta gatgaggacg atgttgacac
     1621 cgtggtagat gaacagccaa agtttatgga atggtgtgct gaggaggaga accaagagct
     1681 catcgccaac ttcaatgccc agtacatgaa agttcagaag ggctggatcc agttggagaa
     1741 agaaggacag ccaacaccaa gagcaaggaa caaatcagat aaactgaaag agatttggaa
     1801 aagcaagaaa aggtcacgga aatgtaggag ttcattggag agtcagaagt gttctcctgt
     1861 tcagatgctc tttatgacaa actttaaatt atctaatgtt tgtaaatggt tcttagagac
     1921 aactgaaacc cggtctctag tcattgtgaa gaagctcaat actcgccttc caggagacgt
     1981 tccccctgtc aagcatcctc ttcagaaata cgctccttcc agcctatatc ccagttcact
     2041 acaggctgag cgcttgaaaa agcacttgaa gaaatttcct ggagctaccc ctgctaagaa
     2101 ttgataatct agagcctgca gtctcgaggc atgcggtacc aagcttgtcg agaagtacta
     2161 gaggatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     2221 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     2281 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     2341 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     2401 gatctgatca ctgatatcgc ctaggagatc cgaaccagat aagtgaaatc tagttccaaa
     2461 ctattttgtc atttttaatt ttcgtattag cttacgacgc tacacccagt tcccatctat
     2521 tttgtcactc ttccctaaat aatccttaaa aactccattt ccacccctcc cagttcccaa
     2581 ctattttgtc cgcccacagc ggggcatttt tcttcctgtt atgtttttaa tcaaacatcc
     2641 tgccaactcc atgtgacaaa ccgtcatctt cggctacttt ttctctgtca cagaatgaaa
     2701 atttttctgt catctcttcg ttattaatgt ttgtaattga ctgaatatca acgcttattt
     2761 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2821 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     2881 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2941 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3001 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3061 agtccacgtt cttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3121 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3181 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
     3241 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3301 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3361 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3421 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3481 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3541 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3601 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3661 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     3721 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     3781 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3841 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     3901 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     3961 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     4021 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     4081 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     4141 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     4201 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     4261 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4321 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4381 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4441 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4501 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4561 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4621 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4681 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4741 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4801 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4861 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4921 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4981 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctc acatgttctt
     5041 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     5101 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     5161 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tagaccagcc
     5221 gcgtaacctg gcaaaatcgg ttacggttga gtaataaatg gatgccctgc gtaagcgggt
     5281 gtgggcggac aataaagtct taaactgaac aaaatagatc taaactatga caataaagtc
     5341 ttaaactaga cagaatagtt gtaaactgaa atcagtccag ttatgctgtg aaaaagcata
     5401 ctggactttt gttatggcta aagcaaactc ttcattttct gaagtgcaaa ttgcccgtcg
     5461 tattaaagag gggcgtggcc aagggcatgg taaagactat attcgcggcg ttgtgacaat
     5521 ttaccgaaca actccgcggc cgggaagccg atctcggctt gaacgaattg ttaggtggcg
     5581 gtacttgggt cgatatcaaa gtgcatcact tcttcccgta tgcccaactt tgtatagaga
     5641 gccactgcgg gatcgtcacc gtaatctgct tgcacgtaga tcacataagc accaagcgcg
     5701 ttggcctcat gcttgaggag attgatgagc gcggtggcaa tgccctgcct ccggtgctcg
     5761 ccggagactg cgagatcata gatatagatc tcactacgcg gctgctcaaa cttgggcaga
     5821 acgtaagccg cgagagcgcc aacaaccgct tcttggtcga aggcagcaag cgcgatgaat
     5881 gtcttactac ggagcaagtt cccgaggtaa tcggagtccg gctgatgttg ggagtaggtg
     5941 gctacgtctc cgaactcacg accgaaaaga tcaagagcag cccgcatgga tttgacttgg
     6001 tcagggccga gcctacatgt gcgaatgatg cccatacttg agccacctaa ctttgtttta
     6061 gggcgactgc cctgctgcgt aacatcgttg ctgctgcgta acatcaaaca tcgacccacg
     6121 gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa
     6181 caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac
     6241 cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc
     6301 aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc
     6361 agcgaagtcg aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg
     6421 catcgtcagg cattggcggc cttgctgttc ttctacggca aggtgc
//
