LOCUS       sequence-346514-        4462 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast MoClo level-0 part plasmid type 234r containing toxic gene 
            dropout cassette (ccdB and TPK2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4462)
  AUTHORS   Otto M, Skrekas C, Gossing M, Gustafsson J, Siewers V, David F
  TITLE     Expansion of the Yeast Modular Cloning Toolkit for CRISPR-Based 
            Applications, Genomic Integrations and Combinatorial Libraries.
  JOURNAL   ACS Synth Biol. 2021 Dec 17;10(12):3461-3474. doi: 
            10.1021/acssynbio.1c00408. Epub 2021 Dec 3.
  PUBMED    34860007
REFERENCE   2  (bases 1 to 4462)
  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: 
            "10.1021/acssynbio.1c00408"; journalName: "ACS Synth Biol"; date: 
            "2021-12-17- 17"; volume: "10"; issue: "12"; pages: "3461-3474"
COMMENT     Sequence Label: pMC234r-ccdB/TPK2
FEATURES             Location/Qualifiers
     source          1..4462
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      120..708
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     609..628
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        831..1238
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             2988..3293
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3172..3191
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     terminator      complement(3636..3670)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(3741..4400)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4315..4334
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tttcattatg gtgaaagttg gaacctctta cgtgcccgat caatcatgac caaaatccct
       61 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
      121 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
      181 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
      241 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
      301 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
      361 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
      421 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
      481 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
      541 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
      601 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
      661 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
      721 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
      781 ttatcccctg attctgtgga taaccgtagt cggtcacaaa cgtgagaccc ccacacacca
      841 tagcttcaaa atgtttctac tcctttttta ctcttccaga ttttctcgga ctccgcgcat
      901 cgccgtacca cttcaaaaca cccaagcaca gcatactaaa tttcccctct ttcttcctct
      961 agggtgtcgt taattacccg tactaaaggt ttggaaaaga aaaaagacac cgcctcgttt
     1021 ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt cttgaaaatt
     1081 tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt taataaacgg
     1141 tcatcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt tttttacttc
     1201 ttgctcatta gaaagaaagc atagcaatct aatctaagtt ttaattacaa aagatctatg
     1261 gaattcgttg cagaaagggc tcagccagtt ggtcaaacaa tccagcagca aaatgttaat
     1321 acttacgggc aaggcgtcct acaaccgcat catgatttac agcagcgaca acaacaacaa
     1381 cagcagcgtc agcatcaaca actgctgacg tcacagttgc cccagaaatc tctcgtatcc
     1441 aaaggcaaat atacactaca tgacttccag attatgagaa cgcttggtac tggatctttt
     1501 ggtagggttc atttggtgcg ctctgttcac aatggtcggt attatgctat aaaagtttta
     1561 aagaaacaac aagttgtcaa gatgaaacag gttgaacata ccaatgacga acgacgtatg
     1621 ctaaagcttg tggagcatcc gtttctgatt agaatgtggg gtacgtttca agatgctagg
     1681 aatatcttta tggtgatgga ttatatcgaa ggtggtgaac ttttctcgtt actgagaaag
     1741 tcacaaagat ttcctaatcc tgtagcaaaa ttttacgctg cggaagtcat actggcatta
     1801 gagtacttgc atgctcataa tatcatctac agggatttaa agccagaaaa tatcttgctg
     1861 gatagaaatg gccacattaa aataaccgat tttgggttcg ccaaagaggt acaaactgtc
     1921 acatggacgc tttgtgggac tcctgattac attgctcctg aagttattac cacaaagcca
     1981 tataataagt cggtggattg gtggtcccta ggtgttctaa tctacgaaat gttagctggt
     2041 tatacaccct tttacgatac tactccaatg aagacgtatg aaaagatttt acaaggtaaa
     2101 gtagtatatc caccatattt tcatcccgac gtcgtggatc tactaagcaa actgatcact
     2161 gcagatttga caagaagaat cggtaattta caaagcggtt ccagggacat caaagctcac
     2221 ccatggttta gtgaagttgt atgggaaaga ttgttagcaa aggatattga aactccatac
     2281 gagcctccta tcacatcagg tatcggtgac acgtctttat tcgatcaata tcccgaggag
     2341 cagctagatt atggtattca aggcgatgat ccatatgctg aatactttca agatttctaa
     2401 ggatcctaac tcgagataaa gcaatcttga tgaggataat gatttttttt tgaatataca
     2461 taaatactac cgtttttctg ctagattttg tgatgacgta aataagtaca tattactttt
     2521 taagccaaga caagattaag cattaacttt acccttttct ttctaagttt caatattagt
     2581 tatcactgtt taaaagttat ggcgagaacg tcggcggtta aaatatatta ccctgaacgg
     2641 ctgacgccgc gtggatccgg cttactaaaa gccagataac agtatgcgta tttgcgcgct
     2701 gatttttgcg gtataagaat atatactgat atgtataccc gaagtatgtc aaaaagaggt
     2761 atgctatgaa gcagcgtatt acagtgacag ttgacagcga cagctatcag ttgctcaagg
     2821 catatatgat gtcaatatct ccggtctggt aagcacaacc atgcagaatg aagcccgtcg
     2881 tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt
     2941 tattgaaatg aacggctctt ttgctgacga gaacaggggc tggtgaaatg cagtttaagg
     3001 tttacaccta taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta
     3061 ttgacacgcc cgggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata
     3121 aagtctcccg tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga
     3181 ccaccgatat ggccagtgtg ccggtttccg ttatcgggga agaagtggct gatctcagcc
     3241 accgcgaaaa tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag
     3301 gctcccttat acacagccag tctgcaggtc gatacagtag aaattacaga aactttatca
     3361 cgtttagtaa gtatagaggc tgaaaatcca gatgaagccg aacgacttgt aagagaaaag
     3421 tataagagtt gtgaaattgt tcttgatgca gatgattttc aggactatga cactagcgta
     3481 tatgaatagg tagatgtttt tattttgtca cacaaaaaag aggctcgcac ctctttttct
     3541 tatttctttt tatgatttaa tacggcattg aggacaatag cgagtaggct gcgatcgcac
     3601 gcattccgtt gggtctcagc tgtgtgacca gaccaataaa aaacgcccgg cggcaaccga
     3661 gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc
     3721 caagcgagct cgatatcaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat
     3781 tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc
     3841 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg
     3901 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg
     3961 gctgaaacga aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg
     4021 taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca
     4081 ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca
     4141 ctatcccata tcaccagctc accgtctttc attgccatac gaaattccgg atgagcattc
     4201 atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg
     4261 gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact
     4321 gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat
     4381 ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa
     4441 aatacgcccg gtagtgatct ta
//
