LOCUS       sequence-489823-        6020 bp    DNA     circular SYN 20-OCT-2025
DEFINITION  PHF19 KO donor plasmid with blasticidin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6020)
  AUTHORS   Much C, Rajkumar SM, Chen L, Cohen JM, Gade AR, Pitt GS, Long Y
  TITLE     Distinct specificity and functions of PRC2 subcomplexes in human 
            stem cells and cardiac differentiation.
  JOURNAL   Mol Cell. 2025 Aug 7:S1097-2765(25)00615-X. doi: 
            10.1016/j.molcel.2025.07.014.
  PUBMED    40803325
REFERENCE   2  (bases 1 to 6020)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 20, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell. 
            2025 Aug 7:S1097-2765(25)00615-X. doi: 
            10.1016/j.molcel.2025.07.014."
COMMENT     Sequence Label: pUC19_PHF19_KO_BSD_donor
FEATURES             Location/Qualifiers
     source          1..6020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1207..1328
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1329..1450
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1451..1572
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1575..1686
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    1789..1822
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1823..2333
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2252..2270
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2343..2741
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    2782..3006
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    3007..3040
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3799..3815)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3799..3815)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3812..3834)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3823..3839
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3847..3853,3854..3871,3872..3877))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3892..3913
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4030..4047)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4201..4789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4281..4300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4960..5751,5752..5820))
                     /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     5583..5602
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5821..5925)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5993..6011
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctctaa ttgagaacag
      421 aaggggggat tacaaaagac agatgaccca gtctacacta gctacaagcc acccagtttt
      481 cctttctagg tgaattttct ctggacagat ctcaggttag ggatggttgc aggggttggg
      541 ggtaaaaaca aaacccaact gcttgcctgg agaagatcct ccatccagag cacatggact
      601 gtgtttgctt aaggaataaa acctcagcct cctgagacag ggagtggttc aggcccgctg
      661 ggtaggggac tttgttctgt gtgggttgag agatccattc tctttaaaag gtagtgtggt
      721 atgggccatg gctctgcctt tggcccatca gcctcttctc cttgccacac ctgtttccct
      781 atctgtaaga tgtgaataat aataatgtct ccctcatagg atggtcacga ggaatcagtg
      841 agataataac caagtctggc ttatagcaca gggtacagaa atggtggttt ccttccttcc
      901 cctgatgggc tttctgaagc agggaagcaa gaagcaaaaa ccctgttctt tctctcccag
      961 tgtcagggga agctgatgga gaatcgagct ctggatccag ggactcggga ctcctatggt
     1021 gccaccagcc acctccccaa caagggggcc ctggcgaagg tcaagaacaa cttcaaagac
     1081 ttgatgtcca aactgacgga gggccagtat gtgctgtgcc ggtggacaga tggcctgtac
     1141 tacctcggga agatcaattg atcttcccga ggtagtacag caagaggcac tggcggggca
     1201 gcaggcaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     1261 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     1321 gtatcttaaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     1381 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     1441 atgtatctta aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     1501 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     1561 caatgtatct tagactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     1621 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     1681 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     1741 caagggggag gattgggaag agaatagcag gcatgcggcc gcgatatcat aacttcgtat
     1801 aatgtatgct atacgaagtt atggggttgg ggttgcgcct tttccaaggc agccctgggt
     1861 ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa acgcagcggc gccgaccctg
     1921 ggtctcgcac attcttcacg tccgttcgca gcgtcacccg gatcttcgcc gctacccttg
     1981 tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg
     2041 cggcgtgccg gacgtgacaa acggaagccg cacgtctcac tagtaccctc gcagacggac
     2101 agcgccaggg agcaatggca gcgcgccgac cgcgatgggc tgtggccaat agcggctgct
     2161 cagcagggcg cgccgagagc agcggccggg aaggggcggt gcgggaggcg gggtgtgggg
     2221 cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag cctccggagc
     2281 gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga cctctctccc cagcaattca
     2341 ccatggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa
     2401 tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg
     2461 gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg
     2521 tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg
     2581 gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc
     2641 tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga
     2701 ttcgtgaatt gctgccctct ggttatgtgt gggagggcta agctgactcg agtctagacc
     2761 aattggttta aaccctgcag gctgtgcctt ctagttgcca gccatctgtt gtttgcccct
     2821 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     2881 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     2941 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     3001 ctatggataa cttcgtataa tgtatgctat acgaagttat cctaggtggc ccctgttctc
     3061 agactcagaa tctctctgct catgttccag cctggctgtc agaccagggg gcagggggat
     3121 tgctggagaa ttcccttctt gtccctcatg cccaaagctg gtggtgggag gggccagagc
     3181 ctggtcttgc aggcaccatc cctagggtca acatcatcca actagttctt tagaaaagtg
     3241 ctctgagagc ctacaaaaca aaaaagtcag gtttgagagg atcacagagg agcatctgac
     3301 ttgtgcttct gtcctcaaca ttctgaccaa gttgccatcc atcggtccct ggctgtacga
     3361 ctccaggtga ctaggagtac accctgtcca ggaactgcca tccttcttcc ccttctagcc
     3421 ctgctttctg gaacatacgg agacaaaaat ggaggaggaa gactatgttc caggttggct
     3481 tttccccagc cctttctcac ttgctccttc ctatctgtac cccaggtcag cagctctaag
     3541 caaagctgcc tcgtgacttt cgaagataat tccaaatact gggtcctatg gaaggacata
     3601 cagcatggtg agttttccta acatttcggg tagcccccaa cctccccagg tcttctctcc
     3661 tctgctgcct tcatttcttc cttctgtcct tccatctctt actcctggca tctgcccttg
     3721 accatgaacc ccaagcccac ctgtgatgat gatcaaatct ggttgtatat tgacttagga
     3781 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3841 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3901 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3961 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4021 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4081 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4141 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4201 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4261 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4321 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4381 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4441 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4501 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4561 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4621 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4681 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4741 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4801 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4861 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4921 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4981 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5041 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5101 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5161 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5221 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5281 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5341 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5401 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5461 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5521 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5581 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5641 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5701 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5761 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5821 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5881 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5941 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     6001 tatcacgagg ccctttcgtc
//
