LOCUS       sequence-214943-        2789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sub cloning vector position 1.
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.HRF_T7_term_HR1
FEATURES             Location/Qualifiers
     source          1..2789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(355..372)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(489..1034)
                     /direction=LEFT
                     /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     complement(569..588)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1234..1253
                     /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     1326..1345
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1326..1344
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1376..1398
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1525..1543)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1539..1586
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     protein_bind    1659..1678
                     /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     1699..1716
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1700..1716
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1990..2649)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2564..2583
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2650..2752)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 cggtatcaac agggacacca ggatttattt attctgcgaa gtgatcttcc gtcacaggta
       61 tttattcggc gcaaagtgcg tcgggtgatg ctgccaactt actgatttag tgtatgatgg
      121 tgtttttgag gtgctccagt ggcttctgtt tctatcagct gtccctcctg ttcagctact
      181 gacggggtgg tgcgtaacgg caaaagcacc gccggacatc agcgctagcg gagtgtatac
      241 tggcttacta tgttggcact gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa
      301 aaaggctgca ccggtgcgtc agcagaatat gtgatacagg atatattccg cttcctcgct
      361 cactgactcg ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt acgaacgggg
      421 cggagatttc ctggaagatg ccaggaagat acttaacagg gaagtgagag ggccgcggca
      481 aagccgtttt tccataggct ccgcccccct gacaagcatc acgaaatctg acgctcaaat
      541 cagtggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggcggctcc
      601 ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg tcattccgct gttatggccg
      661 cgtttgtctc attccacgcc tgacactcag ttccgggtag gcagttcgct ccaagctgga
      721 ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc ttatccggta actatcgtct
      781 tgagtccaac ccggaaagac atgcaaaagc accactggca gcagccactg gtaattgatt
      841 tagaggagtt agtcttgaag tcatgcgccg gttaaggcta aactgaaagg acaagttttg
      901 gtgactgcgc tcctccaagc cagttacctc ggttcaaaga gttggtagct cagagaacct
      961 tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga gcaagagatt acgcgcagac
     1021 caaaacgatc tcaagaagat catcttatta atcagataaa atatttctag atttcagtgc
     1081 aatttatctc ttcaaatgta gcacctgaag tcagccccat acgatataag ttgtaagccc
     1141 tgcaggtcca ggacccgggt ctagacttca tttccttctc cagattaact caattccagc
     1201 atttagaacc ctaccaagta tcgatggccg gccaattgtg agcgctcaca atttcgagct
     1261 ctgagcccta ctgtcgagaa atatgtatct cgcactcgag tggcgcgccc tcgatcccgc
     1321 gaaattaata cgactcacta tagggagacc acaacggttt ccctctagaa ataattttgt
     1381 ttaactttaa gaaggagata tacatatgcg ccggcggcgg ccgcaccttt gtcttcaata
     1441 agggcgatta attaaaggcc tgtttaaact tattgaagac ttaagccctg caggggcgcg
     1501 cctaattaac ctaggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     1561 ctaaacgggt cttgaggggt tttttgctga aaggcgccgg cggcggccgc ggccggccac
     1621 cagcagcttg ccagctcttt cagtatcatg gagcccataa ttgtgagcgc tcacaattac
     1681 tcgagtcccg ggggatcctg taaaacgacg gccagtgaat tctagagcaa taacccatca
     1741 atctgtacgc acatcgatac ctttgtctaa gggcgatacg tacatttgag aagcacacgg
     1801 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt
     1861 aacgaccctg ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc tgccattcat
     1921 ccgcttatta tcacttattc aggcgtagca accaggcgtt taagggcacc aataactgcc
     1981 ttaaaaaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat
     2041 tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag
     2101 caccttgtcg ccttgcgtat aatatttgcc catagtgaaa acgggggcga agaagttgtc
     2161 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa
     2221 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac
     2281 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga
     2341 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat
     2401 caccagctca ccgtctttca ttgccatacg gaactccgga tgagcattca tcaggcgggc
     2461 aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa
     2521 ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc
     2581 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt
     2641 tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     2701 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca
     2761 ttttcgccaa aagttggccc agggcttcc
//
