LOCUS       sequence-214951-        2789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Subcloning vector position 3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2789)
  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 2789)
  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: pACY.HR2_T7_term_HR3
FEATURES             Location/Qualifiers
     source          1..2789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      20..565
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     466..485
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     682..699
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1091..1193
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1194..1853
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1260..1279)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    2165..2184
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     terminator      complement(2257..2304)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2300..2318
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             2445..2467
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2498..2517)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2499..2517)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2590..2609
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
ORIGIN
        1 gattaataag atgatcttct tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa
       61 acgaaaaaac cgccttgcag ggcggttttt cgaaggttct ctgagctacc aactctttga
      121 accgaggtaa ctggcttgga ggagcgcagt caccaaaact tgtcctttca gtttagcctt
      181 aaccggcgca tgacttcaag actaactcct ctaaatcaat taccagtggc tgctgccagt
      241 ggtgcttttg catgtctttc cgggttggac tcaagacgat agttaccgga taaggcgcag
      301 cggtcggact gaacgggggg ttcgtgcata cagtccagct tggagcgaac tgcctacccg
      361 gaactgagtg tcaggcgtgg aatgagacaa acgcggccat aacagcggaa tgacaccggt
      421 aaaccgaaag gcaggaacag gagagcgcac gagggagccg ccagggggaa acgcctggta
      481 tctttatagt cctgtcgggt ttcgccacca ctgatttgag cgtcagattt cgtgatgctt
      541 gtcagggggg cggagcctat ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta
      601 agtatcttcc tggcatcttc caggaaatct ccgccccgtt cgtaagccat ttccgctcgc
      661 cgcagtcgaa cgaccgagcg tagcgagtca gtgagcgagg aagcggaata tatcctgtat
      721 cacatattct gctgacgcac cggtgcagcc ttttttctcc tgccacatga agcacttcac
      781 tgacaccctc atcagtgcca acatagtaag ccagtataca ctccgctagc gctgatgtcc
      841 ggcggtgctt ttgccgttac gcaccacccc gtcagtagct gaacaggagg gacagctgat
      901 agaaacagaa gccactggag cacctcaaaa acaccatcat acactaaatc agtaagttgg
      961 cagcatcacc cgacgcactt tgcgccgaat aaatacctgt gacggaagat cacttcgcag
     1021 aataaataaa tcctggtgtc cctgttgata ccgggaagcc ctgggccaac ttttggcgaa
     1081 aatgagacgt tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac
     1141 taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct aaaatggaga
     1201 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
     1261 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
     1321 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
     1381 ttgcccgcct gatgaatgct catccggagt tccgtatggc aatgaaagac ggtgagctgg
     1441 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
     1501 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
     1561 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
     1621 ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
     1681 tggacaactt cttcgccccc gttttcacta tgggcaaata ttatacgcaa ggcgacaagg
     1741 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
     1801 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg taattttttt
     1861 aaggcagtta ttggtgccct taaacgcctg gttgctacgc ctgaataagt gataataagc
     1921 ggatgaatgg cagaaattcg aaagcaaatt cgacccggtc gtcggttcag ggcagggtcg
     1981 ttaaatagcc gcttatgtct attgctggtt taccggttta ttgactaccg gaagcagtgt
     2041 gaccgtgtgc ttctcaaatg tacgtatcgc ccttagacaa aggtatcgat ggaatcagaa
     2101 gaacgccata ttacagagat agtgatgtga tccgggaatg agaaggtacc cccgggactc
     2161 gagtaattgt gagcgctcac aattatgggc tccatgatac tgaaagagct ggcaagctgc
     2221 tggtggccgg ccgcggccgc cgccggcgcc tttcagcaaa aaacccctca agacccgttt
     2281 agaggcccca aggggttatg ctagttattg ctcagcggtg gcagcagcct aggttaatta
     2341 ggcgcgcccc tgcagggctt aagtcttcaa taagtttaaa caggccttta attaatcgcc
     2401 cttattgaag acaaaggtgc ggccgccgcc ggcgcatatg tatatctcct tcttaaagtt
     2461 aaacaaaatt atttctagag ggaaaccgtt gtggtctccc tatagtgagt cgtattaatt
     2521 tcgcgggatc gagggcgcgc cactcgagtg cgagatacat atttctcgac agtagggctc
     2581 agagctcgaa attgtgagcg ctcacaattg gccggccatc gatctggaaa ggcatactct
     2641 agcaatctgt aggtgatcga cttggaaaac cctggcgaag cttcccgggt cctggacctg
     2701 cagggcttac aacttatatc gtatggggct gacttcaggt gctacatttg aagagataaa
     2761 ttgcactgaa atctagaaat attttatct
//
