LOCUS       sequence-296108-        5319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a targeting plasmid for Nanog-APEX.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5319)
  AUTHORS   Kurihara M, Kato K, Sanbo C, Shigenobu S, Ohkawa Y, Fuchigami T, 
            Miyanari Y
  TITLE     Genomic Profiling by ALaP-Seq Reveals Transcriptional Regulation by 
            PML Bodies through DNMT3A Exclusion.
  JOURNAL   Mol Cell. 2020 May 7;78(3):493-505.e8. doi: 
            10.1016/j.molcel.2020.04.004. Epub 2020 Apr 29.
  PUBMED    32353257
REFERENCE   2  (bases 1 to 5319)
  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"; doi: 
            "10.1016/j.molcel.2020.04.004"; journalName: "Mol Cell"; date: 
            "2020-05-7- 7"; volume: "78"; issue: "3"; pages: "493-505.e8"
COMMENT     Sequence Label: pGbH500-Nanog-AXP
FEATURES             Location/Qualifiers
     source          1..5319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..87)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     136..158
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     150..167
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     151..167
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             748..774
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1753..1809
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1816..2415
                     /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(1816..1835)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2312..2332
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2428..2444)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2428..2444)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3399..4190
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      4283..4871
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4772..4791
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5025..5042
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5201..5228)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
       61 ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc
      121 tagcatggat gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagcgtctcc
      181 ccctgaacct gaccccggac caagtggtgg gaattcatgg cgagggcgat gccacctacg
      241 gaacggcctg tgcggccatg ctaatctttt cagtatttct gcagtttaca tctttagagt
      301 ccccttttca ctcattgcat taatcacctc tttcagaagg gctcagcacc agtggagtat
      361 cccagcatcc attgcagcta tccccagggc tatctggtga acgcatctgg aagcctttcc
      421 atgtggggca gccagacttg gaccaaccca acttggagca gccagacctg gaccaaccca
      481 acttggaaca accagacctg gaccaaccca acttggagca gccaggcctg gaccgctcag
      541 tcctggaacg gccagccttg gaatgctgct ccgctccata acttcgggga ggactttctg
      601 cagccttacg tacagttgca gcaaaacttc tctgccagtg atttggaggt gaatttggaa
      661 gccactaggg aaagccatgc gcattttagc accccacaag ccttggaatt attcctgaac
      721 tactctgtga ctccaccagg tgaaatatac ccatacgatg ttccagatta cgctggtgga
      781 ggcggaggtt ctgggggagg aggtagtggc ggtggtggtt caggaggcgg cggaagcttg
      841 gatccaggtg gaggtggaag cggtagcaaa ggaacgcgtg gcaaatcata tcctacagtg
      901 tctgccgact atcaggacgc tgtggagaaa gctaagaaga aactgagagg cttcatcgct
      961 gagaagcggt gtgcccccct gatgctgaga ctggctttcc acagcgccgg aaccttcgac
     1021 aagggaacca agacaggcgg ccctttcgga accatcaagc accccgctga gctcgcccat
     1081 agcgctaaca acggactgga catcgccgtg cggctgctgg agcctctgaa ggccgagttc
     1141 cctatcctga gctatgccga cttctaccag ctggctggag tggtggccgt ggaggttacc
     1201 ggaggaccta aggtgccctt ccaccctgga agagaggaca agcctgagcc tccacccgag
     1261 ggacggctgc ctgatcctac caagggaagc gaccatctgc gggacgtgtt cggaaaggcc
     1321 atgggactga ccgatcaaga catcgtggcc ctcagcggcg gacatacaat cggagctgct
     1381 cacaaggaga gaagcggctt cgaaggcccc tggacaagca accccctgat cttcgacaac
     1441 agctacttta ccgaactgct cagcggcgag aaggagggac tgttgcagct gcccagcgac
     1501 aaggctctgc tcagcgaccc tgtgttcaga cccctggtgg acaagtacgc tgctgacgag
     1561 gatgccttct tcgctgacta cgccgaggct caccagaagc tgagcgagct gggcttcgct
     1621 gacgctctgc agatcgataa gcttgaggtg cacaccaacc aggaccccct ggacgccgaa
     1681 gtccatacaa atcaggatcc tctggatgcc gaagtgcaca ccaatcagga tcccctggac
     1741 gctggaagcg gagctactaa cttcagcctg ctgaagcagg ctggtgacgt ggaggagaac
     1801 cctggacctg aattcatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     1861 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     1921 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     1981 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     2041 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     2101 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     2161 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     2221 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     2281 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     2341 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     2401 aagcccggtg cctgatgcag cccgggggat ccactagttc tagagcggcc gaaatatgag
     2461 acttacgcaa catctgggct taaagtcagg gcaaagccag gttccttcct tcttccaaat
     2521 attttcatat tttttttaaa gatttattta ttcattatat gtaagtacac tgtagctgtc
     2581 ttcagacact ccagaagagg gcgtcagatc ttgttacgta tggttgtgag ccaccatgtg
     2641 gttgctggga tttgaactcc tgaccttcgg aagagcagtc gggtgctctt atccactgag
     2701 ccatctcacc agcccctggt ttattttttt aattattatt tgctttttgt ttatcaagac
     2761 agggtttctc tgcatagctc taattgtctt tgaactagct ctgcagacca gcctggcctt
     2821 gaactcagag atctgcccac ttatctttgc ctcctgaatg ctgggaccaa aggtggcata
     2881 ccaccacacc tggcatatat attgtttatt tctatttcta tttttattgg tgccagagca
     2941 aacctaggac ccgtaggtgg catcgccctc gccgcaagca agcgctcgaa acggtgcagc
     3001 ggctgttgcc ggtgctgtgc caggaccatg gccgagacgt ctagaccagc caggacagaa
     3061 atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg aaacggatga
     3121 aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta
     3181 tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg
     3241 cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca
     3301 agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta
     3361 cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg gtgatcgccg
     3421 aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt
     3481 tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata
     3541 ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca
     3601 acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag
     3661 tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc
     3721 aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg
     3781 acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc
     3841 cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg
     3901 aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc
     3961 ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg
     4021 ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta
     4081 gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag
     4141 aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc
     4201 acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg
     4261 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4321 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4381 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4441 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4501 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4561 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4621 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4681 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4741 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4801 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4861 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4921 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     4981 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5041 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5101 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5161 atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat
     5221 ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc
     5281 gggccgttgc ttcacaacgt tcaaatccgc tcccggcgg
//
