LOCUS       sequence-210639-        3713 bp    DNA     circular SYN 15-APR-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 Apr 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    138..171
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     378..397
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        378..396
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             428..450
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(477..496)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             1451..1468
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1505..1552
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1646..1693
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1756..1778)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1864..2003
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1919..1938
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2018..2035)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2189..2777)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2269..2288)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2808..2847)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     promoter        complement(join(3712..3713,1..90))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
       61 tatttgaatg tatttagaaa aataaacaaa taggggaatt aaaaaaaagc ccgctcatta
      121 ggcgggctac tagtcagata acttcgtata atgtatgcta tacgaagtta tggtaccgcg
      181 gccgcgtaga ggatctgttg atcagcagtt caacctgttg atagtacttc gttaatacag
      241 atgtaggtgt tggcaccatg cataactata acggtcctaa ggtagcgacc taggttctag
      301 acttcatttc cttctccaga ttaactcaat tccagcattt agaaccctac caagtatcga
      361 tctcgatccc gcgaaattaa tacgactcac tatagggaga ccacaacggt ttccctctag
      421 aaataatttt gtttaacttt aagaaggaga tatacatatg aaatatgtaa cgttatacga
      481 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
      541 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
      601 tcccaaccgc gtggcacaac aactggcggg caaacagctg ttgctgattg gcgttgccac
      661 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
      721 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
      781 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
      841 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
      901 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
      961 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     1021 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
     1081 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
     1141 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
     1201 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat
     1261 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac
     1321 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
     1381 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac
     1441 caccggctct caccatcacc atcaccatta attaacctag gctgctgcca ccgctgagca
     1501 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     1561 ctcgagagat ccggctgcta acaaacccgc ttaaggaaga tggccaactt atttacataa
     1621 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     1681 gaggggtttt ttggtttaaa cccatctaat tgggaattcc ccctgcctcg cgcgtttcgg
     1741 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     1801 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     1861 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg
     1921 gcatcagagc agattgtact gagagtgcac cattgcggtg tgaaataccg cacagatgcg
     1981 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2041 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2101 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2161 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2221 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2281 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2341 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2401 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2461 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2521 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2581 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     2641 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2701 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2761 gaaaaaaagg atctcaagaa gatcctttga tcttttctac cggtccgtcc aaaaaaaaag
     2821 gctccaaaag gagcctttaa ttgtatcggt ttaccaatgc ttaatcagtg aagcccctat
     2881 ttcagcaatc tgtctatttc gttcgtccat agttgcctga ctacccgtcg tgtagataac
     2941 tacgatacgg gaaggcttac catctggtcc aagtgcggca atgatacccc gtgaaccacg
     3001 ctctcctgcc ccagatttat cagcaataaa ccagccagca ggaagtgccg aacgcagtag
     3061 tggtcctgca actttatccg cttcaagcca gtctattaat tgttgccgtg aagccagagt
     3121 aagtagttcg ccagttaata gtttgcgtaa tgttgttgcc attgccgcag gcattgtggt
     3181 gtctcgctcg tcgtttggta tggcttcatt cagttctggt tcccaacggt caaggcgagt
     3241 tacatggtct cccatattgt gcaaaaaagc ggttaattcc ttcggtccac cgatggttgt
     3301 cagaagtaag ttcgccgcag tgttatcact catcgttatg gcggcactac ataattctct
     3361 taccgtcatt ccgtccgtaa ggtgcttttc tgtcactggg gagtattcaa ccaagtcatt
     3421 ctgcgaatag tgtattcgtc gtccgagttg ctcttgcccc gcatctacac gggataatac
     3481 tgctccacat agcagaactt tgaaagtgct catcattgga aaacgctctt cagggcgaaa
     3541 actctcaagg attttaccgc tattcaagtc cagttctatg tatcccactc gtgcccccaa
     3601 ttggtcctct gcgtctttta ctttcacgag cgtttctggg tgagcaaaaa caggaaggca
     3661 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tac
//
