LOCUS       sequence-227423-        7936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7936)
  AUTHORS   Nishio H, Walsh MJ
  TITLE     CCAAT displacement protein/cut homolog recruits G9a histone lysine 
            methyltransferase to repress transcription.
  JOURNAL   Proc Natl Acad Sci U S A. 2004 Aug 3;101(31):11257-62. Epub 2004 Jul
            21.
  PUBMED    15269344
REFERENCE   2  (bases 1 to 7936)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2004-08-3"; volume: "101(31)"; pages: 
            "11257-62. Epub 2004 Jul 21"
COMMENT     Sequence Label: pGL2-p21 promoter-Luc
FEATURES             Location/Qualifiers
     source          1..7936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2414..4066
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(2480..2498)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          4307..4372
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4502..4522
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4947..5081
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5223..5240)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5394..5982)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5474..5493)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6153..6944,6945..7013))
                     /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     6776..6795
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7014..7118)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7145..7600
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7232..7251)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7442..7463
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    7790..7911
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 cccgggaggt accgagctct tacgcgtgct agctcgagat ctaagtaagc ttgggcagca
       61 ggctgtggct ctgattggct ttctggccat caggaacatg tcccaacatg ttgagctctg
      121 gcatagaaga ggctggtggc tattttgtcc ttgggctgcc tgtttccagg tgaggaaggg
      181 gatggtagga gacaggagac ctctaaagac cccaggtaaa ccttagcctg ttactctgaa
      241 cagggtatgt gatctgccag cagatccttg cgacagggct gggatctgat gcatgtgtgc
      301 ttgtgtgagt gtgtgctggg agtcagattc tgtgtgtgac ttttaacagc ctgctccctt
      361 gccttcttca gggcagaagt cctcccttag agtgtgtctg ggtacacatt caagtgcatg
      421 gttgcaaact ttttttttta aagcactgaa tagtactaga cacttagtag gtacttaaga
      481 aatattgaat gtcgtggtgg tggtgagcta gaagttataa aaaaaattct ttcccaaaaa
      541 caacaacaaa aagaattatt tcattgtgaa gctcagtacc acaaaaattt aaataattca
      601 ttacaagcct tcattaaaaa atattttctc cccaaagtaa acagacagac aatgtctagt
      661 ctatttgaaa tgcctgaaag cagaggggct tcaaggcagt gggagaaggt gcctgtcctc
      721 tgctggacat ttgacaacca gccctttgga tggtttgtat gtataggagc gaaggtgcag
      781 acagcagtgg ggcttagagt ggggtcctga ggctgtgcta tggcccttct ggggtttagc
      841 cacaatcctg gcctgactcc agggcgaggc aggccaaggg ggtctgctgc tgtgtcctcc
      901 cacccctacc tgggctccca tccccacagc agaggagaaa gaagcctgtc ctccccgagg
      961 tcagctgcgt tagaggaaga agactgggca tgtctgggca gagatttcca gactctgtgc
     1021 agcctgagat gtcagtaatt gtagctgctc caagcctggg ttctgttttt cagtgggatt
     1081 tctgttcaga tgaacaatcc atcctctgca attttttaaa agcaaaactg caaatgtttc
     1141 aggcacagaa aggaggcaaa ggtgaagtcc aggggaggtc aggggtgtga ggtagatggg
     1201 agcggataga cacatcactc atttctgtgt ctgtcagaag aaccagtaga cacttccaga
     1261 attgtccttt atttatgtca tctccataaa ccatctgcaa atgagggtta tttggcattt
     1321 ttgtcatttt ggaaccacag aaataaagga tgacaagcag agagccccgg gcaggaggca
     1381 aaagtcctgt gttccaacta tagtcatttc tttgctgcat gatctgagtt aggtcaccag
     1441 acttctctga gccccagtct ccccagcagt gtatacgggc tatgtgggga gtattcagga
     1501 gacagacaac tcactcgtca aatcctcccc ttcctggcca acaaagctgc tgcaaccaca
     1561 ggggtttctt ctgttcaggt gagtgtaggg tgtagggaga ttggttcaat gtccaattct
     1621 tctgtttccc tggagatcag gttgcccttt tttggtagtc tctccaattc cctccttccc
     1681 ggaagcatgt gacaatcaac aactttgtat acttaagttc agtggacctc aatttcctca
     1741 tctgtgaaat aaacgggact gaaaaatcat tctggcctca agatgctttg ttggggtgtc
     1801 taggtgctcc aggtgcttct gggagaggtg acctagtgag ggatcagtgg gaatagaggt
     1861 gatattgtgg ggcttttctg gaaattgcag agaggtgcat cgtttttata atttatgaat
     1921 ttttatgtat taatgtcatc ctcctgatct tttcagctgc attgggtaaa tccttgcctg
     1981 ccagagtggg tcagcggtga gccagaaagg gggctcattc taacagtgct gtgtcctcct
     2041 ggagagtgcc aactcattct ccaagtgaaa agagccagat ttgtggctca cttcgtgggg
     2101 aaatgtgtcc agcgcaccaa cgcaggtgag ggactggggg aggagggaag tgccctcctg
     2161 cagcacgcga ggttccggga ccggctggcc tgctggaact cggccaggct cagctggctc
     2221 cgcgctgggc agccaggagc ctgggccccg gggagggcgg tcccgggcgg cgcggtgggc
     2281 cgagcgcggg tcccgcctcc ttgaggcggg cccgggcggg gcggttgtat atcagggccg
     2341 cgctgagctg cgccagctga ggtgtgagca gctgccgaag tcagaagctt ggcattccgg
     2401 tactgttggt aaaatggaag acgccaaaaa cataaagaaa ggcccggcgc cattctatcc
     2461 tctagaggat ggaaccgctg gagagcaact gcataaggct atgaagagat acgccctggt
     2521 tcctggaaca attgctttta cagatgcaca tatcgaggtg aacatcacgt acgcggaata
     2581 cttcgaaatg tccgttcggt tggcagaagc tatgaaacga tatgggctga atacaaatca
     2641 cagaatcgtc gtatgcagtg aaaactctct tcaattcttt atgccggtgt tgggcgcgtt
     2701 atttatcgga gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa
     2761 cagtatgaac atttcgcagc ctaccgtagt gtttgtttcc aaaaaggggt tgcaaaaaat
     2821 tttgaacgtg caaaaaaaat taccaataat ccagaaaatt attatcatgg attctaaaac
     2881 ggattaccag ggatttcagt cgatgtacac gttcgtcaca tctcatctac ctcccggttt
     2941 taatgaatac gattttgtac cagagtcctt tgatcgtgac aaaacaattg cactgataat
     3001 gaattcctct ggatctactg ggttacctaa gggtgtggcc cttccgcata gaactgcctg
     3061 cgtcagattc tcgcatgcca gagatcctat ttttggcaat caaatcattc cggatactgc
     3121 gattttaagt gttgttccat tccatcacgg ttttggaatg tttactacac tcggatattt
     3181 gatatgtgga tttcgagtcg tcttaatgta tagatttgaa gaagagctgt ttttacgatc
     3241 ccttcaggat tacaaaattc aaagtgcgtt gctagtacca accctatttt cattcttcgc
     3301 caaaagcact ctgattgaca aatacgattt atctaattta cacgaaattg cttctggggg
     3361 cgcacctctt tcgaaagaag tcggggaagc ggttgcaaaa cgcttccatc ttccagggat
     3421 acgacaagga tatgggctca ctgagactac atcagctatt ctgattacac ccgaggggga
     3481 tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg ttgtggatct
     3541 ggataccggg aaaacgctgg gcgttaatca gagaggcgaa ttatgtgtca gaggacctat
     3601 gattatgtcc ggttatgtaa acaatccgga agcgaccaac gccttgattg acaaggatgg
     3661 atggctacat tctggagaca tagcttactg ggacgaagac gaacacttct tcatagttga
     3721 ccgcttgaag tctttaatta aatacaaagg atatcaggtg gcccccgctg aattggaatc
     3781 gatattgtta caacacccca acatcttcga cgcgggcgtg gcaggtcttc ccgacgatga
     3841 cgccggtgaa cttcccgccg ccgttgttgt tttggagcac ggaaagacga tgacggaaaa
     3901 agagatcgtg gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt
     3961 tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag
     4021 agagatcctc ataaaggcca agaagggcgg aaagtccaaa ttgtaaaatg taactgtatt
     4081 cagcgatgac gaaattctta gctattgtaa tactgcgatg agtggcaggg cggggcgtaa
     4141 tttttttaag gcagttattg gtgcccttaa acgcctggtt gctacgcctg aataagtgat
     4201 aataagcgga tgaatggcag aaattcgccg gatctttgtg aaggaacctt acttctgtgg
     4261 tgtgacataa ttggacaaac tacctacaga gatttaaagc tctaaggtaa atataaaatt
     4321 tttaagtgta taatgtgtta aactactgat tctaattgtt tgtgtatttt agattccaac
     4381 ctatggaact gatgaatggg agcagtggtg gaatgccttt aatgaggaaa acctgttttg
     4441 ctcagaagaa atgccatcta gtgatgatga ggctactgct gactctcaac attctactcc
     4501 tccaaaaaag aagagaaagg tagaagaccc caaggacttt ccttcagaat tgctaagttt
     4561 tttgagtcat gctgtgttta gtaatagaac tcttgcttgc tttgctattt acaccacaaa
     4621 ggaaaaagct gcactgctat acaagaaaat tatggaaaaa tattctgtaa cctttataag
     4681 taggcataac agttataatc ataacatact gttttttctt actccacaca ggcatagagt
     4741 gtctgctatt aataactatg ctcaaaaatt gtgtaccttt agctttttaa tttgtaaagg
     4801 ggttaataag gaatatttga tgtatagtgc cttgactaga gatcataatc agccatacca
     4861 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     4921 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     4981 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5041 gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgtcgaccg atgcccttga
     5101 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
     5161 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctcttccgct
     5221 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5281 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5341 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5401 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5461 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5521 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5581 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5641 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5701 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5761 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5821 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5881 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5941 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6001 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6061 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6121 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6181 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6241 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6301 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6361 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6421 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6481 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6541 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6601 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6661 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6721 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6781 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6841 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6901 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6961 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7021 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7081 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7141 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     7201 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     7261 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     7321 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     7381 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     7441 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     7501 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     7561 tttaacgcga attttaacaa aatattaacg cttacaattt gccattcgcc attcaggctg
     7621 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gcccaagcta
     7681 ccatgataag taagtaatat taaggtacgt ggaggtttta cttgctttaa aaaacctccc
     7741 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     7801 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     7861 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atggtactgt
     7921 aactgagcta acataa
//
