LOCUS       sequence-214954-        2789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Subcloning vector position 5.
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.HR4_T7_term_HRR
FEATURES             Location/Qualifiers
     source          1..2789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             100..122
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(153..172)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(154..172)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    245..264
                     /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."
     rep_origin      464..1009
                     /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     910..929
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1126..1143
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1535..1637
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1638..2297
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1704..1723)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    2609..2628
                     /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(2701..2748)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2744..2762
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 gcccctgcag ggcttaagtc ttcaataagt ttaaacaggc ctttaattaa tcgcccttat
       61 tgaagacaaa ggtgcggccg ccgccggcgc atatgtatat ctccttctta aagttaaaca
      121 aaattatttc tagagggaaa ccgttgtggt ctccctatag tgagtcgtat taatttcgcg
      181 ggatcgaggg cgcgccactc gagtgcgaga tacatatttc tcgacagtag ggctcagagc
      241 tcgaaattgt gagcgctcac aattggccgg ccatcgatgc tcagaaccta cactaacgac
      301 attgggtagc gacgtaacta acagctctta caaatctccc cgggtcctgg acctgcaggg
      361 cttacaactt atatcgtatg gggctgactt caggtgctac atttgaagag ataaattgca
      421 ctgaaatcta gaaatatttt atctgattaa taagatgatc ttcttgagat cgttttggtc
      481 tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt ttttcgaagg
      541 ttctctgagc taccaactct ttgaaccgag gtaactggct tggaggagcg cagtcaccaa
      601 aacttgtcct ttcagtttag ccttaaccgg cgcatgactt caagactaac tcctctaaat
      661 caattaccag tggctgctgc cagtggtgct tttgcatgtc tttccgggtt ggactcaaga
      721 cgatagttac cggataaggc gcagcggtcg gactgaacgg ggggttcgtg catacagtcc
      781 agcttggagc gaactgccta cccggaactg agtgtcaggc gtggaatgag acaaacgcgg
      841 ccataacagc ggaatgacac cggtaaaccg aaaggcagga acaggagagc gcacgaggga
      901 gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc accactgatt
      961 tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa acggctttgc
     1021 cgcggccctc tcacttccct gttaagtatc ttcctggcat cttccaggaa atctccgccc
     1081 cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga gtcagtgagc
     1141 gaggaagcgg aatatatcct gtatcacata ttctgctgac gcaccggtgc agcctttttt
     1201 ctcctgccac atgaagcact tcactgacac cctcatcagt gccaacatag taagccagta
     1261 tacactccgc tagcgctgat gtccggcggt gcttttgccg ttacgcacca ccccgtcagt
     1321 agctgaacag gagggacagc tgatagaaac agaagccact ggagcacctc aaaaacacca
     1381 tcatacacta aatcagtaag ttggcagcat cacccgacgc actttgcgcc gaataaatac
     1441 ctgtgacgga agatcacttc gcagaataaa taaatcctgg tgtccctgtt gataccggga
     1501 agccctgggc caacttttgg cgaaaatgag acgttgatcg gcacgtaaga ggttccaact
     1561 ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag
     1621 gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc
     1681 aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc
     1741 agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt
     1801 tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg gagttccgta
     1861 tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt
     1921 tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc
     1981 agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc
     2041 ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca
     2101 gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc actatgggca
     2161 aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg
     2221 tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt
     2281 ggcagggcgg ggcgtaattt ttttaaggca gttattggtg cccttaaacg cctggttgct
     2341 acgcctgaat aagtgataat aagcggatga atggcagaaa ttcgaaagca aattcgaccc
     2401 ggtcgtcggt tcagggcagg gtcgttaaat agccgcttat gtctattgct ggtttaccgg
     2461 tttattgact accggaagca gtgtgaccgt gtgcttctca aatgtacgta tcgcccttag
     2521 acaaaggtat cgatttatgt aaataagttg gccatcttcc ttaagcgggt ttgttagcag
     2581 ccggatctct cgagcccagg actcgagtaa ttgtgagcgc tcacaattat gggctccatg
     2641 atactgaaag agctggcaag ctgctggtgg ccggccgcgg ccgccgccgg cgcctttcag
     2701 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctagtt attgctcagc
     2761 ggtggcagca gcctaggtta attaggcgc
//
