LOCUS       sequence-198368-        3447 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express DNA binding domain of mouse CTCF in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3447)
  AUTHORS   Zuo Z, Roy B, Chang YK, Granas D, Stormo GD
  TITLE     Measuring quantitative effects of methylation on transcription 
            factor-DNA binding affinity.
  JOURNAL   Sci Adv. 2017 Nov 17;3(11):eaao1799. doi: 10.1126/sciadv.aao1799. 
            eCollection 2017 Nov.
  PUBMED    29159284
REFERENCE   2  (bases 1 to 3447)
  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: "Sci Adv."; 
            date: "2017-11-17"; volume: "3(11):eaao1799. doi"; pages: " 
            10.1126/sciadv.aao1799. eCollection 2017 Nov"
COMMENT     Sequence Label: hisSUMO-CTCF(F1-F9)
FEATURES             Location/Qualifiers
     source          1..3447
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             53..75
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             86..103
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             122..412
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIG"
     terminator      1434..1481
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(1699..2287)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1779..1798)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2459..3250,3251..3319))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRVHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3082..3101
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3320..3424)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tggtgctagc cccgcgaaat taatacgact cactataggg tctagaaata attttgttta
       61 actttaagaa ggagatatac atatgcatca ccatcaccat cacggttctt ctatggctag
      121 catgtcggac tcagaagtca atcaagaagc taagccagag gtcaagccag aagtcaagcc
      181 tgagactcac atcaatttaa aggtgtccga tggatcttca gagatcttct tcaagatcaa
      241 aaagaccact cctttaagaa ggctgatgga agcgttcgct aaaagacagg gtaaggaaat
      301 ggactcctta agattcttgt acgacggtat tagaattcaa gctgatcaga cccctgaaga
      361 tttggacatg gaggataacg atattattga ggctcacaga gaacagattg gtctggtgcc
      421 aagcggtagt aatgcagaga aagtggttgg taatatgaag cctccgaagc caacaaaaat
      481 taaaaaaaaa ggtgtaaaga aaacattcca gtgtgagctt tgcagttaca catgtccccg
      541 gcgttcaaat ttggatcgtc acatgaaaag ccacactgat gagagaccac acaaatgcca
      601 tctgtgtggc agagcattca gaacagtgac cctcctgagg aatcatctga acacacacac
      661 aggtactcgt cctcacaagt gcccagactg cgatatggcc tttgtgacca gtggagaatt
      721 ggtgcggcat cgtcgttata aacacactca tgagaaacca tttaagtgtt ccatgtgtga
      781 ttatgccagt gtagaagtca gcaaattaaa acgacacatt cgctctcata ctggagagcg
      841 cccgttccag tgcagtttgt gcagttatgc cagcagggac acatacaagc tgaaaaggca
      901 tatgagaacc cattcagggg aaaaacctta tgaatgttat atttgtcacg ctcggtttac
      961 ccagagtggt accatgaaga tgcacatttt acagaagcac acagaaaatg tggccaaatt
     1021 tcattgtccc cattgtgaca ctgtcatagc ccgaaaaagt gatttgggtg tccacttgcg
     1081 aaagcagcat tcctatattg aacagggcaa aaaatgtcgc tactgtgatg ctgtgtttca
     1141 tgagcgatat gctctcatcc agcatcaaaa atcacacaaa aatgagaagc gcttcaagtg
     1201 tgaccagtgt gattatgctt gtagacagga gcggcacatg atcatgcaca agcgcactca
     1261 cacgggggag aagccttatt aatgaggatc ccgggaattc tcgagtaagg ttaacctgca
     1321 ggaggccttt aattaaggtg gtgcggccgc gctagcggtc ccgggggatc gatccggctg
     1381 ctaacaaagc ccgaaaggaa gctgagttag ctgctgccac cgctgaacaa taactagcat
     1441 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     1501 ccggaagctt ggcactggcc gaccggggtc gagcactgac tcgctgcgct cggtcgttcg
     1561 gccgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1621 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1681 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1741 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1801 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1861 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1921 ggtgtgggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1981 ctgcgcctta tccggtaact atcgtcttga gtccaacccg ctaagacacg acttatcgcc
     2041 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2101 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2161 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2221 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2281 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2341 acagatccgg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2401 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2461 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     2521 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     2581 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     2641 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     2701 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     2761 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     2821 gctccggttc ccaacgatca aggcgggtta catgatcccc catgttgtgc aaaaaagcgg
     2881 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     2941 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3001 tgactggtga gtactcaacc aagtcattct gagaatagtg tacgcggcga ccgagttgct
     3061 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3121 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3181 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3241 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3301 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3361 attgtctcat gagcggatac atatttgaat gcatttagaa aaataaacaa ataggggttc
     3421 cgcgcacatt tccccgaaaa gtgctag
//
