LOCUS       sequence-489759-        6001 bp    DNA     circular SYN 20-OCT-2025
DEFINITION  PHF1 KO donor plasmid with blasticidin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6001)
  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 6001)
  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_PHF1_KO_BSD_donor
FEATURES             Location/Qualifiers
     source          1..6001
                     /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    1172..1293
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1294..1415
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1416..1537
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1540..1651
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    1754..1787
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1788..2298
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2217..2235
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2308..2706
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    2747..2971
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    2972..3005
                     /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(3780..3796)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3780..3796)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3793..3815)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3804..3820
                     /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(3828..3834,3835..3852,3853..3858))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3873..3894
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4011..4028)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4182..4770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4262..4281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4941..5732,5733..5801))
                     /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     5564..5583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5802..5906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5974..5992
                     /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 cgagctctgg aagctgggaa
      421 actagcaact aggtttgggg tcggggggag ggctgttagt taccgagagg agagccagtc
      481 actagagttg gaagaggact caaaaagctg gtgatgggtg cccagtggtt tcctatcaag
      541 gtgtctcagg gatttggaga tgggggcaca gatacttcat tcttgatggc aaaataggca
      601 aagaaaatgg ggaggctggc cgggcccggt ggctcacgcc tgtaatccca gcactttggg
      661 aagctgaggt gggtggatca tgaggtcagg agttcgagac cagtttggcc aacatggcca
      721 acacctcatc tctactaaaa atacaaaaat tagccaggca tggtggcatg cgcctgtaat
      781 cccacctact tgggagactg aggcaggaga attgcctgaa cccgggaggt ggaggttgca
      841 gtgagccaag attgtgccac tgcactctgg cctgggcgac agagcaaaac tccatctcag
      901 gaaaaaaaaa aaaaaaagaa aaagaaaatg gggaaggttt ctcagcattc ataaccttct
      961 cctccccatt tcttttctgg ctaggccccc ccaggatgca atggcgcagc ccccccggct
     1021 gagccgctct ggtgcctcct cactttggga cccagcttct cctgctccca cctctggccc
     1081 caggcctcgg ctttgggagg gtcaagatgt gctggccaga tggactgatg ggctgctata
     1141 cttgggtacc acctttgaac aatcaggtac caacttgttt attgcagctt ataatggtta
     1201 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     1261 ttgtggtttg tccaaactca tcaatgtatc ttaaacttgt ttattgcagc ttataatggt
     1321 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1381 agttgtggtt tgtccaaact catcaatgta tcttaaactt gtttattgca gcttataatg
     1441 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     1501 ctagttgtgg tttgtccaaa ctcatcaatg tatcttagac tgtgccttct agttgccagc
     1561 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     1621 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     1681 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagagaat agcaggcatg
     1741 cggccgcgat atcataactt cgtataatgt atgctatacg aagttatggg gttggggttg
     1801 cgccttttcc aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc
     1861 gggaaacgca gcggcgccga ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc
     1921 acccggatct tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta
     1981 agtcgggaag gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt
     2041 ctcactagta ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga
     2101 tgggctgtgg ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg
     2161 gcggtgcggg aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt
     2221 tccgcattct gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc
     2281 accgacctct ctccccagca attcaccatg gccaagcctt tgtctcaaga agaatccacc
     2341 ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc
     2401 gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt
     2461 actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc
     2521 aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga
     2581 cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt
     2641 gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag
     2701 ggctaagctg actcgagtct agaccaattg gtttaaaccc tgcaggctgt gccttctagt
     2761 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2821 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2881 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2941 aggcatgctg gggatgcggt gggctctatg gataacttcg tataatgtat gctatacgaa
     3001 gttatcctag ggtaagacct tctacctctg accttcttcc tagttccctt atctaattct
     3061 ggttcccatt tcatcctgtt tgctcaccca ttccaggtgg acagtgctag ggaggtgtgt
     3121 ctggtccagt ttgaggatga ttcgcagttt ctggttctat ggaaagacat tagccctggt
     3181 aagactctag agacctgaga ttgcacatcc catggaaaac aaacctggcc tagaggggca
     3241 gaaagagact taaaggcagg ccctgtgaca ctgtgttctc tcacagctgc cctccctgga
     3301 gaggaactcc tctgttgtgt ctgtcgctct gagactgtgg tccctgggaa ccggctggtc
     3361 agctgtgaga agtgtcgcca tggtgagagg gcaggtcacc tgaatggtcc agcttgctct
     3421 tccctccagg atggtctcta tatcacctgt cctggcctta gtccccaagc cactgctctg
     3481 gccaggctgc ttagccttat cttagtcctc atccgctttc agcccaggga gaagcagcca
     3541 tggaaagggg tggtataacc tttgcaggca gaagatggtt taaatgcatc cttttctaac
     3601 catatgacct cggacaagtc ccttaacctc taggagcctc agttccctga cttgtaaaat
     3661 aaggataatg caccccctca tcaagaccat ggtcagggat taaatgagat gggtaaagac
     3721 tattcctgtc ccaataccaa gcacacacag gtatgcaata aaagcttggc gtaatcatgg
     3781 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3841 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3901 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3961 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4021 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4081 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4141 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4201 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4261 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4321 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4381 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4441 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4501 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4561 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4621 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4681 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4741 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4801 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4861 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4921 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4981 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5041 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5101 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5161 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5221 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5281 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5341 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5401 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5461 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5521 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5581 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5641 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5701 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5761 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5821 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5881 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     5941 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     6001 c
//
