LOCUS       sequence-194643-        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     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(1095..1113)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1109..1156
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(1374..1962)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1454..1473)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2134..2925,2926..2994))
                     /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     2757..2776
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2995..3099)
                     /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 gtagtagaga ggagaagcct gagcctgagt ctgtgtatga gacagactgc
      481 cgctgggatg gttgcagcca ggagttcgat tcccaggagc agctggtgca ccacatcaac
      541 agtgagcata tccacgggga gcggaaggaa ttcgtgtgcc attggggagg ttgctccagg
      601 gagctgaggc ccttcaaggc ccaatacatg ctggtggtgc acatgcgcag acacacgggc
      661 gagaagccac acaagtgcac gtttgaaggc tgtcggaagt cctattcacg ccttgaaaac
      721 ctcaagacgc accttcggtc gcacacgggt gagaagcctt acatgtgtga gcaagaaggt
      781 tgcagcaagg cctttagcaa tgccagtgac cgcgccaagc accagaatcg gacccactcc
      841 aatgagaagc catacgtgtg caagctcccc ggctgcacca agcgctacac agatcccagc
      901 tcgctccgca aacacgtgaa gacagtgcat ggtccggatg cccacgtgac caagtaatga
      961 ggatcccggg aattctcgag taaggttaac ctgcaggagg cctttaatta aggtggtgcg
     1021 gccgcgctag cggtcccggg ggatcgatcc ggctgctaac aaagcccgaa aggaagctga
     1081 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     1141 cttgaggggt tttttgctga aaggaggaac tatatccgga agcttggcac tggccgaccg
     1201 gggtcgagca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1261 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1321 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1381 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1441 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1501 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     1561 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     1621 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     1681 cttgagtcca acccgctaag acacgactta tcgccactgg cagcagccac tggtaacagg
     1741 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     1801 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     1861 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1921 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1981 tctacggggt ctgacgctca gtggaacgaa aactcacaga tccgggattt tggtcatgag
     2041 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2101 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2161 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2221 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2281 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     2341 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2401 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     2461 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     2521 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     2581 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     2641 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     2701 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     2761 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     2821 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     2881 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     2941 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3001 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3061 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagt
//
