LOCUS       sequence-375873-        7788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cell line generation via recombination-mediated cassette exchange 
            (RMCE) and stable expression of CBX7.Chromo/TAF3.PHD-13XL-BASU.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7788)
  AUTHORS   Villasenor R, Pfaendler R, Ambrosi C, Butz S, Giuliani S, Bryan E, 
            Sheahan TW, Gable AL, Schmolka N, Manzo M, Wirz J, Feller C, von 
            Mering C, Aebersold R, Voigt P, Baubec T
  TITLE     ChromID identifies the protein interactome at chromatin marks.
  JOURNAL   Nat Biotechnol. 2020 Mar 2. pii: 10.1038/s41587-020-0434-2. doi: 
            10.1038/s41587-020-0434-2.
  PUBMED    32123383
REFERENCE   2  (bases 1 to 7788)
  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 
            Biotechnol. 2020 Mar 2. pii: 10.1038/s41587-020-0434-2. doi: 
            10.1038/s41587-020-0434-2."
COMMENT     Sequence Label: pv9_CBX7_Chromo/TAF3.PHD-13XL-BASU
FEATURES             Location/Qualifiers
     source          1..7788
                     /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"
     protein_bind    409..442
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     enhancer        447..826
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          1104..2120
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2044..2066
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2128..2147
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2198..2207
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2207..2230
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2528..2548
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2891..2911
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3197..3223
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    4101..4563
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4157..4174)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4384..4403
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4577..5176
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4577..4596)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5073..5093
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5345..5466
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5382..5401)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5436..5455
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(5500..5533)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     primer_bind     complement(5567..5583)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5567..5583)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5580..5602)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5591..5607
                     /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(5615..5621,5622..5639,5640..5645))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5660..5681
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5798..5815)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5969..6557)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6049..6068)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6728..7519,7520..7588))
                     /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     7351..7370
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7589..7693)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7761..7779
                     /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 ctcgagccat aacttcgtat
      421 aatgtatact atacgaagtt atatcggaca ttgattattg actagttatt aatagtaatc
      481 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      541 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      601 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      661 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      721 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      781 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
      841 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
      901 tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggsgc gcgccrggsg
      961 gggggggggg gggsgggggg gggggsgggg sgrggcggag aggtgcggcg gcagccaatc
     1021 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata
     1081 aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct
     1141 ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga
     1201 gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg
     1261 tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg
     1321 ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg
     1381 cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg
     1441 tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgagggga
     1501 acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg
     1561 tcgggctgca accccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg
     1621 gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc
     1681 aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggagggctcg ggggaggggc
     1741 gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt
     1801 tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga
     1861 aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg
     1921 gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc
     1981 ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg
     2041 cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg
     2101 ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt
     2161 ttggcaaaga attcattatt gtttttcctc ctttgccgcc accatggact acaaagacga
     2221 tgacgacaag gatgaagagt ctggttctgt ttctggtggt gatgagctgt cagccatagg
     2281 cgagcaggtg tttgcggtgg agagcatccg gaagaagcgc gtgcggaagg gcaaagttga
     2341 atatctggtg aagtggaaag gatggccccc caagtatagc acctgggagc cagaggagca
     2401 catcttggac cctcgccttg tcatggccta cgaggagaag gaggagagag accgagcctc
     2461 ggggtatagg aagagaggtc cgaaacccat cgttggcgga ggctctggag gtggaggctc
     2521 cgcacccccc aaaaagaaac gcaaggttgg ataccgggat ccaggcgggg ggtctagtgc
     2581 aaaggcacct gtccgaagtg tcgtcacaga aactgtgtct acttatgtca tcagagacga
     2641 atggggaaat cagatttgga tctgtcccgg ctgcaataag ccagatgacg gatcacctat
     2701 gataggatgt gacgactgcg atgactggta tcactggcct tgcgttggga tcatggccgc
     2761 cccccccgaa gagatgcaat ggttttgccc caaatgcgcc aataaaataa aaaaggacaa
     2821 gaaacataag aaacgcaagc atcgcgctca catcggtggc ggaggctctg gaggtggagg
     2881 ctccgcaccc cccaaaaaga aacgcaaggt tggataccgg gatccacacg tgggtaggag
     2941 cggttcgttc ggatcccatc ttttttcgga attcggatcc ggtggaggcg ggtctggagg
     3001 cgggggtagt ggcgggggtg gaagcggggg tggaggcggg tcgggtggcg gaggtagcgg
     3061 aggcggtgga agtggaggcg gtggaagtgg tggaggaggt tctggtggcg gaggtagcgg
     3121 tggcggaggt agcggaggcg gtggaagtgg tggcggaggt agcggaggcg gtggatcggc
     3181 gcgccgtttc aagaactacc catacgatgt gcctgactat gctggcaagt tgtctgaatc
     3241 agagattagg tttggactga aaacggaggt gatgggccaa catcttatct atcacgatgt
     3301 tttgtccagc acacaaaaga cagcccatga attggcaaat aacaatgccc ccgagggtac
     3361 cctcgtggtt gcagataaac agacagccgg taggggcggg atgagccgag tgtggcattc
     3421 tcaagaagga aacggagtat ggatgagttt gatattgcgc ccggacattc cgctccaaaa
     3481 gacgccccaa cttaccttgc tggcagctgt agcggtggtt cagggaatag aagaggccgc
     3541 gggcatacag actgatataa agtggccgaa cgacatactc attaacggga aaaaaacagt
     3601 agggatactt accgaaatgc aagctgagga ggatcgagtg cgctccgtga ttattggtat
     3661 tggcattaat gtaaatcagc aacccaatga tttccccgat gagctcaagg acattgcgac
     3721 gagtctgtcc caggcagctg gggaaaagat cgacagggca ggtgtaatcc agcatatctt
     3781 gctttgtttc gaaaagcgat accgagacta catgacccac ggtttcacgc ccatcaagct
     3841 cttgtgggag tcatacgccc tcggtattgg cacgaacatg agggcgagaa ctctcaacgg
     3901 gacattttac ggtaaggcgt tgggtattga cgacgaaggg gtacttttgt tggaaactaa
     3961 tgaaggcatt aagaagatat acagcgcgga catcagcctt aggtgaaacg aggtagcggc
     4021 cgccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
     4081 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
     4141 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
     4201 gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
     4261 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
     4321 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
     4381 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
     4441 tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta
     4501 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
     4561 taatatggcc acaaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     4621 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     4681 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     4741 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     4801 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     4861 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     4921 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     4981 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     5041 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     5101 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     5161 caagcccggt gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcagt
     5221 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5281 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5341 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5401 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5461 atcttatcat gtctgtatac cgtggatccg gccggcccca taacttcgta tagtatacat
     5521 tatacgaagt tatcccctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc
     5581 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5641 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5701 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5761 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5821 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5881 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5941 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6001 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6061 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6121 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6181 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6241 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6301 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6361 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6421 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6481 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6541 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6601 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6661 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6721 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6781 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6841 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6901 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6961 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7021 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7081 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7141 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7201 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7261 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7321 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7381 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7441 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7501 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7561 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7621 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7681 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     7741 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
