LOCUS       sequence-198359-        3117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express hisSUMO tagged DNA binding domain of Gli1(mouse) in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3117)
  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 3117)
  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-Gli1
FEATURES             Location/Qualifiers
     source          1..3117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     22..41
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        22..40
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             54..76
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             87..104
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             123..413
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGREMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIG"
     CDS             414..431
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      1105..1152
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(1370..1958)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1450..1469)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2130..2921,2922..2990))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLVDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQAAMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2753..2772
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2991..3095)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtggtgctag ccccgcgaaa ttaatacgac tcactatagg gtctagaaat aattttgttt
       61 aactttaaga aggagatata catatgcatc accatcacca tcacggttct tctatggcta
      121 gcatgtcgga ctcagaagtc aatcaagaag ctaagccaga ggtcaagcca gaagtcaagc
      181 ctgagactca catcaattta aaggtgtccg atggatcttc agagatcttc ttcaagatca
      241 aaaagaccac tcctttaaga aggctgatgg aagcgttcgc taaaagacag ggtagggaaa
      301 tggactcctt aagattcttg tacgacggta ttagaattca agctgatcag acccctgaag
      361 atttggacat ggaggataac gatattattg aggctcacag agaacagatt ggtctggtgc
      421 cacgcggtag tagagaggag aagcctgagc ctgagtctgt gtatgagaca gactgccgct
      481 gggatggttg cagccaggag ttcgattccc aggagcagct ggtgcaccac atcaacagtg
      541 agcatatcca cggggagcgg aaggaattcg tgtgccattg gggaggttgc tccagggagc
      601 tgaggccctt caaggcccaa tacatgctgg tggtgcacat gcgcagacac acgggcgaga
      661 agccacacaa gtgcacgttt gaaggctgtc ggaagtccta ttcacgcctt gaaaacctca
      721 agacgcacct tcggtcgcac acgggtgaga agccttacat gtgtgagcaa gaaggttgca
      781 gcaaggcctt tagcaatgcc agtgaccgcg ccaagcacca gaatcggacc cactccaatg
      841 agaagccata cgtgtgcaag ctccccggct gcaccaagcg ctacacagat cccagctcgc
      901 tccgcaaaca cgtgaagaca gtgcatggtc cggatgccca cgtgaccaag taatgaggat
      961 cccgggaatt ctcgagtaag gttaacctgc aggaggcctt taattaaggt ggtgcggccg
     1021 cgctagcggt cccgggggat cgatccggct gctaacaaag cccgaaagga agctgagtta
     1081 gctgctgcca ccgctgaaca ataactagca taaccccttg gggcctctaa acgggtcttg
     1141 aggggttttt tgctgaaagg aggaactata tccggaagct tggcactggc cgaccggggt
     1201 cgagcactga ctcgctgcgc tcggtcgttc ggccgcggcg agcggtatca gctcactcaa
     1261 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1321 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1381 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1441 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1501 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1561 ctcatagctc acgctgtagg tatctcagtt cggtgtgggt cgttcgctcc aagctgggct
     1621 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1681 agtccaaccc gctaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1741 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1801 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1861 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1921 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1981 cggggtctga cgctcagtgg aacgaaaact cacagatccg ggattttggt catgagatta
     2041 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2101 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2161 tcagcgatct gtctatttcg ttcatccata gctgcctgac tccccgtcgt gtagataact
     2221 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     2281 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2341 ggtcctgcaa ctttatccgc ctccatccag tctactaatt gttgccggga agctagagta
     2401 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     2461 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     2521 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     2581 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     2641 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     2701 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     2761 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     2821 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     2881 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac gggaaggcaa
     2941 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3001 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3061 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgcta
//
