LOCUS       sequence-214949-        2789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Subcloning vector position 2.
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.HR1_T7_term_HR2
FEATURES             Location/Qualifiers
     source          1..2789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             136..158
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(189..208)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(190..208)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    281..300
                     /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."
     primer_bind     complement(351..368)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(351..367)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      500..1045
                     /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     946..965
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1162..1179
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1571..1673
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1674..2333
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1740..1759)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    2645..2664
                     /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(2737..2784)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     join(2780..2789,1..9)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 gctcagcggt ggcagcagcc taggttaatt aggcgcgccc ctgcagggct taagtcttca
       61 ataagtttaa acaggccttt aattaatcgc ccttattgaa gacaaaggtg cggccgccgc
      121 cggcgcatat gtatatctcc ttcttaaagt taaacaaaat tatttctaga gggaaaccgt
      181 tgtggtctcc ctatagtgag tcgtattaat ttcgcgggat cgagggcgcg ccactcgagt
      241 gcgagataca tatttctcga cagtagggct cagagctcga aattgtgagc gctcacaatt
      301 ggccggccat cgatgtgcgt acagattgat gggttattgc tctagaattc actggccgtc
      361 gttttacagg atcccccggg tcctggacct gcagggctta caacttatat cgtatggggc
      421 tgacttcagg tgctacattt gaagagataa attgcactga aatctagaaa tattttatct
      481 gattaataag atgatcttct tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa
      541 acgaaaaaac cgccttgcag ggcggttttt cgaaggttct ctgagctacc aactctttga
      601 accgaggtaa ctggcttgga ggagcgcagt caccaaaact tgtcctttca gtttagcctt
      661 aaccggcgca tgacttcaag actaactcct ctaaatcaat taccagtggc tgctgccagt
      721 ggtgcttttg catgtctttc cgggttggac tcaagacgat agttaccgga taaggcgcag
      781 cggtcggact gaacgggggg ttcgtgcata cagtccagct tggagcgaac tgcctacccg
      841 gaactgagtg tcaggcgtgg aatgagacaa acgcggccat aacagcggaa tgacaccggt
      901 aaaccgaaag gcaggaacag gagagcgcac gagggagccg ccagggggaa acgcctggta
      961 tctttatagt cctgtcgggt ttcgccacca ctgatttgag cgtcagattt cgtgatgctt
     1021 gtcagggggg cggagcctat ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta
     1081 agtatcttcc tggcatcttc caggaaatct ccgccccgtt cgtaagccat ttccgctcgc
     1141 cgcagtcgaa cgaccgagcg tagcgagtca gtgagcgagg aagcggaata tatcctgtat
     1201 cacatattct gctgacgcac cggtgcagcc ttttttctcc tgccacatga agcacttcac
     1261 tgacaccctc atcagtgcca acatagtaag ccagtataca ctccgctagc gctgatgtcc
     1321 ggcggtgctt ttgccgttac gcaccacccc gtcagtagct gaacaggagg gacagctgat
     1381 agaaacagaa gccactggag cacctcaaaa acaccatcat acactaaatc agtaagttgg
     1441 cagcatcacc cgacgcactt tgcgccgaat aaatacctgt gacggaagat cacttcgcag
     1501 aataaataaa tcctggtgtc cctgttgata ccgggaagcc ctgggccaac ttttggcgaa
     1561 aatgagacgt tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac
     1621 taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct aaaatggaga
     1681 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
     1741 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
     1801 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
     1861 ttgcccgcct gatgaatgct catccggagt tccgtatggc aatgaaagac ggtgagctgg
     1921 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
     1981 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
     2041 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
     2101 ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
     2161 tggacaactt cttcgccccc gttttcacta tgggcaaata ttatacgcaa ggcgacaagg
     2221 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
     2281 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg taattttttt
     2341 aaggcagtta ttggtgccct taaacgcctg gttgctacgc ctgaataagt gataataagc
     2401 ggatgaatgg cagaaattcg aaagcaaatt cgacccggtc gtcggttcag ggcagggtcg
     2461 ttaaatagcc gcttatgtct attgctggtt taccggttta ttgactaccg gaagcagtgt
     2521 gaccgtgtgc ttctcaaatg tacgtatcgc ccttagacaa aggtatcgat ctggaaaggc
     2581 atactctagc aatctgtagg tgatcgactt ggaaaaccct ggcgaagctt cccgggactc
     2641 gagtaattgt gagcgctcac aattatgggc tccatgatac tgaaagagct ggcaagctgc
     2701 tggtggccgg ccgcggccgc cgccggcgcc tttcagcaaa aaacccctca agacccgttt
     2761 agaggcccca aggggttatg ctagttatt
//
