LOCUS       sequence-215597-        3713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses LacI_I12X86 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3713)
  AUTHORS   Jenni S, Harrison SC
  TITLE     Structure of the DASH/Dam1 complex shows its role at the yeast 
            kinetochore-microtubule interface.
  JOURNAL   Science. 2018 May 4;360(6388):552-558. doi: 10.1126/science.aar6436.
  PUBMED    29724956
REFERENCE   2  (bases 1 to 3713)
  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"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2018-05-4- 4"; volume: "360"; issue: 
            "6388"; pages: "552-558"
COMMENT     Sequence Label: pAA.3.lacIh_I12X86
FEATURES             Location/Qualifiers
     source          1..3713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    135..168
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     375..394
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        375..393
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(474..493)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             1448..1465
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1502..1549
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1643..1690
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1753..1775)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1861..2000
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1916..1935
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2015..2032)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2186..2774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2266..2285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2805..2844)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     promoter        complement(join(3709..3713,1..87))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
       61 ttgaatgtat ttagaaaaat aaacaaatag gggaattaaa aaaaagcccg ctcattaggc
      121 gggctactag tcagataact tcgtataatg tatgctatac gaagttatgg taccgcggcc
      181 gcgtagagga tctgttgatc agcagttcaa cctgttgata gtacttcgtt aatacagatg
      241 taggtgttgg caccatgcat aactataacg gtcctaaggt agcgacctag gttctagact
      301 tcatttcctt ctccagatta actcaattcc agcatttaga accctaccaa gtatcgatct
      361 cgatcccgcg aaattaatac gactcactat agggagacca caacggtttc cctctagaaa
      421 taattttgtt taactttaag aagaagatat acatatgaaa tatgtaacgt tatacgatgt
      481 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
      541 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
      601 caaccgcgtg gcacaacaac tggcgggcaa acagctgttg ctgattggcg ttgccacctc
      661 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
      721 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc
      781 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
      841 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
      901 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact
      961 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
     1021 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
     1081 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
     1141 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
     1201 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc
     1261 ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat
     1321 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
     1381 gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac
     1441 cggctctcac catcaccatc accattaatt aacctaggct gctgccaccg ctgagcaata
     1501 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggctc
     1561 gagagatccg gctgctaaca aacccgctta aggaagatgg ccaacttatt tacataaggc
     1621 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     1681 gggttttttg gtttaaaccc atctaattgg gaattccccc tgcctcgcgc gtttcggtga
     1741 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     1801 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     1861 cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca
     1921 tcagagcaga ttgtactgag agtgcaccat tgcggtgtga aataccgcac agatgcgtaa
     1981 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2041 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     2101 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2161 gtaaaaaggc cgcgttgctg gcgtttttcc ataggttccg cccccctgac gagcatcaca
     2221 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2281 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2341 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2401 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2461 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2521 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2581 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     2641 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2701 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2761 aaaaaggatc tcaagaagat cctttgatct tttctaccgg tccgtccaaa aaaaaaggct
     2821 ccaaaaggag cctttaattg tatcggttta ccaatgctta atcagtgaag cccctatttc
     2881 agcaatctgt ctatttcgtt cgtccatagt tgcctgacta cccgtcgtgt agataactac
     2941 gatacgggaa ggcttaccat ctggtccaag tgcggcaatg ataccccgtg aaccacgctc
     3001 tcctgcccca gatttatcag caataaacca gccagcagga agtgccgaac gcagtagtgg
     3061 tcctgcaact ttatccgctt caagccagtc tattaattgt tgccgtgaag ccagagtaag
     3121 tagttcgcca gttaatagtt tgcgtaatgt tgttgccatt gccgcaggca ttgtggtgtc
     3181 tcgctcgtcg tttggtatgg cttcattcag ttctggttcc caacggtcaa ggcgagttac
     3241 atggtctccc atattgtgca aaaaagcggt taattccttc ggtccaccga tggttgtcag
     3301 aagtaagttc gccgcagtgt tatcactcat cgttatggcg gcactacata attctcttac
     3361 cgtcattccg tccgtaaggt gcttttctgt cactggggag tattcaacca agtcattctg
     3421 cgaatagtgt attcgtcgtc cgagttgctc ttgccccgca tctacacggg ataatactgc
     3481 tccacatagc agaactttga aagtgctcat cattggaaaa cgctcttcag ggcgaaaact
     3541 ctcaaggatt ttaccgctat tcaagtccag ttctatgtat cccactcgtg cccccaattg
     3601 gtcctctgcg tcttttactt tcacgagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     3661 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tct
//
