LOCUS       sequence-346551-        4842 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  preassembled yeast MoClo level-1 backbone with GFP dropout and URA3 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4842)
  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 4842)
  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: pMC-Ura-Cen-lvl1
FEATURES             Location/Qualifiers
     source          1..4842
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      377..448
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      464..491
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             859..1662
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     misc_feature    1745..2248
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(join(2359..3150,3151..3219))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2982..3001
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3220..3324)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3427..4015
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3916..3935
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     RBS             4476..4487
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(4494..4842,1..365)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
ORIGIN
        1 ataccctggt aaaccgcatt gagctgaaag gcattgactt taaagaggac ggcaatatcc
       61 tgggccataa gctggaatac aattttaaca gccacaatgt ttacatcacc gccgataaac
      121 aaaaaaatgg cattaaagcg aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc
      181 agctggctga tcactaccag caaaacactc caatcggtga tggtcctgtt ctgctgccag
      241 acaatcacta tctgagcacg caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc
      301 atatggttct gctggagttc gtaaccgcag cgggcatcac gcatggtatg gatgaactgt
      361 acaaatgacc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
      421 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
      481 gcctttctgc gtttataggt ctcagctgcc aatgagacga cggggtcatc acggctcatc
      541 atgcgccaaa caaatgtgtg caatacacgc tcggatgact gcatgatgac cgcactgact
      601 ggggacagca gatccaccta agcctgtgag agaagcagac acccgacaga tcaaggcagt
      661 taactagtgc actgcagtac acggtttcct tgaaattttt ttgattcggt aatctccgaa
      721 cagaaggaag aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt
      781 gttgaagaaa catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacgtgc
      841 aggaaacgaa gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct
      901 agtcctgttg ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct
      961 tcattggatg ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa
     1021 atttgtttac taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt
     1081 aagccgctaa aggcattatc cgccaagtac aattttttac tcttcgagga cagaaaattt
     1141 gctgacattg gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa
     1201 tgggcagaca ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag
     1261 caggcggcag aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca
     1321 tgcaagggct ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc
     1381 gacaaagatt ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt
     1441 tacgattggt tgattatgac acccggtgtg ggtttagatg acaagggaga tgcattgggt
     1501 caacagtata gaaccgtgga tgatgtggtt tctacaggat ctgacattat tattgttgga
     1561 agaggactat ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca
     1621 ggctgggaag catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta
     1681 aatgcatgta tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc
     1741 gagtatcacg tgctataaaa ataattataa tttaaatttt ttaatataaa tatataaatt
     1801 aaaaatagaa agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc cgaagatgta
     1861 aaagactcta gggggatcgc caacaaatac taccttttat cttgctcttc ctgctctcag
     1921 gtattaatgc cgaattgttt catcttgtct gtgtagaaaa ccacacacga aaatcctgtg
     1981 attttacatt ttacttatcg ttaatcgaat gtatatctat ttaatctgct tttcttgtct
     2041 aataaatata tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt gtttattttt
     2101 ttttcttcat tccgtaactc ttctaccttc tttatttact ttctaaaatc caaatacaaa
     2161 acataaaaat aaataaacac agagtaaatt cccaaattat tccatcatta aaagatacga
     2221 ggcgcgtgta agttacaggc aagcgatccg tcccgagcgg ccgcgattat caaaaaggat
     2281 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2341 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2401 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2461 gggcttacca tctggcccca gtgctgcaat gataccgcgg gacccacgct caccggctcc
     2521 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     2581 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     2641 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     2701 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     2761 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     2821 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     2881 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     2941 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3001 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3061 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3121 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3181 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3241 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3301 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg tcatgaccaa
     3361 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3421 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3481 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3541 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3601 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3661 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3721 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3781 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3841 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3901 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3961 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4021 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4081 tcctgcgtta tcccctgatt ctgtggataa ccgtgcggcc gcccctgaat tcgcatctag
     4141 atggtagagc cacaaacagc cggtacaagc aacgatctcc aggaccatct gaatcatgcg
     4201 cggatgacac gaactcacga cggcgatcac agacattaac ccacagtaca gacactgcga
     4261 caacgtggca attcgtcgca ataccgtctc actgaactgg ccgataattg cagacgaacg
     4321 tgagaccgaa agtgaaacgt gatttcatgc gtcattttga acattttgta aatcttattt
     4381 aataatgtgt gcggcaattc acatttaatt tatgaatgtt ttcttaacat cgcggcaact
     4441 caagaaacgg caggttcgga tcttagctac tagagaaaga ggagaaatac tagatgcgta
     4501 aaggcgaaga gctgttcact ggtgtcgtcc ctattctggt ggaactggat ggtgatgtca
     4561 acggtcataa gttttccgtg cgtggcgagg gtgaaggtga cgcaactaat ggtaaactga
     4621 cgctgaagtt catctgtact actggtaaac tgccggttcc ttggccgact ctggtaacga
     4681 cgctgactta tggtgttcag tgctttgctc gttatccgga ccatatgaag cagcatgact
     4741 tcttcaagtc cgccatgccg gaaggctatg tgcaggaacg cacgatttcc tttaaggatg
     4801 acggcacgta caaaacgcgt gcggaagtga aatttgaagg cg
//
