LOCUS       sequence-215593-        2789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Subcloning vector position 4.
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.HR3_T7_term_HR4
FEATURES             Location/Qualifiers
     source          1..2789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      10..555
                     /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     456..475
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     672..689
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1081..1183
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1184..1843
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1250..1269)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    2155..2174
                     /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(2247..2294)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2290..2308
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             2435..2457
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2488..2507)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2489..2507)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2580..2599
                     /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 atgatcttct tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac
       61 cgccttgcag ggcggttttt cgaaggttct ctgagctacc aactctttga accgaggtaa
      121 ctggcttgga ggagcgcagt caccaaaact tgtcctttca gtttagcctt aaccggcgca
      181 tgacttcaag actaactcct ctaaatcaat taccagtggc tgctgccagt ggtgcttttg
      241 catgtctttc cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcggact
      301 gaacgggggg ttcgtgcata cagtccagct tggagcgaac tgcctacccg gaactgagtg
      361 tcaggcgtgg aatgagacaa acgcggccat aacagcggaa tgacaccggt aaaccgaaag
      421 gcaggaacag gagagcgcac gagggagccg ccagggggaa acgcctggta tctttatagt
      481 cctgtcgggt ttcgccacca ctgatttgag cgtcagattt cgtgatgctt gtcagggggg
      541 cggagcctat ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta agtatcttcc
      601 tggcatcttc caggaaatct ccgccccgtt cgtaagccat ttccgctcgc cgcagtcgaa
      661 cgaccgagcg tagcgagtca gtgagcgagg aagcggaata tatcctgtat cacatattct
      721 gctgacgcac cggtgcagcc ttttttctcc tgccacatga agcacttcac tgacaccctc
      781 atcagtgcca acatagtaag ccagtataca ctccgctagc gctgatgtcc ggcggtgctt
      841 ttgccgttac gcaccacccc gtcagtagct gaacaggagg gacagctgat agaaacagaa
      901 gccactggag cacctcaaaa acaccatcat acactaaatc agtaagttgg cagcatcacc
      961 cgacgcactt tgcgccgaat aaatacctgt gacggaagat cacttcgcag aataaataaa
     1021 tcctggtgtc cctgttgata ccgggaagcc ctgggccaac ttttggcgaa aatgagacgt
     1081 tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt
     1141 attttttgag ttatcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac
     1201 tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca
     1261 gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa
     1321 gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct
     1381 gatgaatgct catccggagt tccgtatggc aatgaaagac ggtgagctgg tgatatggga
     1441 tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg
     1501 gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg
     1561 ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtctc
     1621 agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt
     1681 cttcgccccc gttttcacta tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc
     1741 gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa
     1801 tgaattacaa cagtactgcg atgagtggca gggcggggcg taattttttt aaggcagtta
     1861 ttggtgccct taaacgcctg gttgctacgc ctgaataagt gataataagc ggatgaatgg
     1921 cagaaattcg aaagcaaatt cgacccggtc gtcggttcag ggcagggtcg ttaaatagcc
     1981 gcttatgtct attgctggtt taccggttta ttgactaccg gaagcagtgt gaccgtgtgc
     2041 ttctcaaatg tacgtatcgc ccttagacaa aggtatcgat gctcagaacc tacactaacg
     2101 acattgggta gcgacgtaac taacagctct tacaaatctc cccgggactc gagtaattgt
     2161 gagcgctcac aattatgggc tccatgatac tgaaagagct ggcaagctgc tggtggccgg
     2221 ccgcggccgc cgccggcgcc tttcagcaaa aaacccctca agacccgttt agaggcccca
     2281 aggggttatg ctagttattg ctcagcggtg gcagcagcct aggttaatta ggcgcgcccc
     2341 tgcagggctt aagtcttcaa taagtttaaa caggccttta attaatcgcc cttattgaag
     2401 acaaaggtgc ggccgccgcc ggcgcatatg tatatctcct tcttaaagtt aaacaaaatt
     2461 atttctagag ggaaaccgtt gtggtctccc tatagtgagt cgtattaatt tcgcgggatc
     2521 gagggcgcgc cactcgagtg cgagatacat atttctcgac agtagggctc agagctcgaa
     2581 attgtgagcg ctcacaattg gccggccatc gatggaatca gaagaacgcc atattacaga
     2641 gatagtgatg tgatccggga atgagaaggt acccccgggt cctggacctg cagggcttac
     2701 aacttatatc gtatggggct gacttcaggt gctacatttg aagagataaa ttgcactgaa
     2761 atctagaaat attttatctg attaataag
//
