LOCUS       sequence-194644-        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     26..45
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        26..44
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             58..80
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             91..108
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..417
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIG"
     CDS             418..435
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1425..1443)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1439..1486
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(1704..2292)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1784..1803)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2464..3255,3256..3324))
                     /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     3087..3106
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3325..3429)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gctagtggtg ctagccccgc gaaattaata cgactcacta tagggtctag aaataatttt
       61 gtttaacttt aagaaggaga tatacatatg catcaccatc accatcacgg ttcttctatg
      121 gctagcatgt cggactcaga agtcaatcaa gaagctaagc cagaggtcaa gccagaagtc
      181 aagcctgaga ctcacatcaa tttaaaggtg tccgatggat cttcagagat cttcttcaag
      241 atcaaaaaga ccactccttt aagaaggctg atggaagcgt tcgctaaaag acagggtaag
      301 gaaatggact ccttaagatt cttgtacgac ggtattagaa ttcaagctga tcagacccct
      361 gaagatttgg acatggagga taacgatatt attgaggctc acagagaaca gattggtctg
      421 gtgccacgcg gtagtaatgc agagaaagta gttggtaata tgaagcctcc gaagccaaca
      481 aaaattaaaa aaaaaggtgt aaagaaaaca ttccagtgtg agctttgcag ttacacatgt
      541 ccccggcgtt caaatttgga tcgtcacatg aaaagccaca ctgatgagag accacacaaa
      601 tgccatctgt gtggcagagc attcagaaca gtgaccctcc tgaggaatca tctgaacaca
      661 cacacaggta ctcgtcctca caagtgccca gactgcgata tggcctttgt gaccagtgga
      721 gaattggtgc ggcatcgtcg ttataaacac actcatgaga aaccatttaa gtgttccatg
      781 tgtgattatg ccagtgtaga agtcagcaaa ttaaaacgac acattcgctc tcatactgga
      841 gagcgcccgt tccagtgcag tttgtgcagt tatgccagca gggacacata caagctgaaa
      901 aggcatatga gaacccattc aggggaaaaa ccttatgaat gttatatttg tcacgctcgg
      961 tttacccaga gtggtaccat gaagatgcac attttacaga agcacacaga aaatgtggcc
     1021 aaatttcatt gtccccattg tgacactgtc atagcccgaa aaagtgattt gggtgtccac
     1081 ttgcgaaagc agcattccta tattgaacag ggcaaaaaat gtcgctactg tgatgctgtg
     1141 tttcatgagc gatatgctct catccagcat caaaaatcac acaaaaatga gaagcgcttc
     1201 aagtgtgacc agtgtgatta tgcttgtaga caggagcggc acatgatcat gcacaagcgc
     1261 actcacacgg gggagaagcc ttattaatga ggatcccggg aattctcgag taaggttaac
     1321 ctgcaggagg cctttaatta aggtggtgcg gccgcgctag cggtcccggg ggatcgatcc
     1381 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     1441 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     1501 tatatccgga agcttggcac tggccgaccg gggtcgagca ctgactcgct gcgctcggtc
     1561 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1621 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1681 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1741 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1801 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1861 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1921 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1981 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccgctaag acacgactta
     2041 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2101 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2161 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2221 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2281 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2341 aactcacaga tccgggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2401 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     2461 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2521 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     2581 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     2641 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     2701 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     2761 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     2821 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     2881 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     2941 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3001 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3061 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     3121 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     3181 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3241 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     3301 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     3361 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3421 ggttccgcgc acatttcccc gaaaagt
//
