LOCUS       sequence-369608-        3292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Assembly cassette with ConL7 and ConRE connectors to be used in the 
            Dueber YTK and Ellis MYT systems.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3292)
  AUTHORS   Shaw WM, Khalil AS, Ellis T
  TITLE     A Multiplex MoClo Toolkit for Extensive and Flexible Engineering of 
            Saccharomyces cerevisiae.
  JOURNAL   ACS Synth Biol. 2023 Nov 6. doi: 10.1021/acssynbio.3c00423.
  PUBMED    37930278
REFERENCE   2  (bases 1 to 3292)
  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"; journalName: "ACS Synth 
            Biol. 2023 Nov 6. doi: 10.1021/acssynbio.3c00423."
COMMENT     Sequence Label: pMYT052
FEATURES             Location/Qualifiers
     source          1..3292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             464..475
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     rep_origin      complement(938..1526)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1018..1037)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1629..1733
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1734..1802,1803..2594)
                     /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     complement(1952..1971)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      complement(2910..2937)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2953..3024)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(3036..3292,1..457))
                     /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 tgatgtaaac attgtggctg ttaaaattgt attccagctt atggcccagg atattgccgt
       61 cctctttaaa gtcaatgcct ttcagctcaa tgcggtttac cagggtatcg ccttcaaatt
      121 tcacttccgc acgcgttttg tacgtgccgt catccttaaa ggaaatcgtg cgttcctgca
      181 catagccttc cggcatggcg gacttgaaga agtcatgctg cttcatatgg tccggataac
      241 gagcaaagca ctgaacacca taagtcagcg tcgttaccag agtcggccaa ggaaccggca
      301 gtttaccagt agtacagatg aacttcagcg tcagtttacc attagttgcg tcaccttcac
      361 cctcgccacg cacggaaaac ttatgaccgt tgacatcacc atccagttcc accagaatag
      421 ggacgacacc agtgaacagt tcttcgcctt tacgcatcta gtatttctcc tctttctcta
      481 gtagctaaga tccgaacctg ccgtttcttg agttgccgcg atgttaagaa aacattcata
      541 aattaaatgt gaattgccgc acacattatt aaataagatt tacaaaatgt tcaaaatgac
      601 gcatgaaatc acgtttcact ttcggtctca cgtttcgttg agacgacact ctagatgtgg
      661 aattcagcac ctttcccacg taaacgcgct cgtcaactgt agacccgaaa atcgtatctg
      721 tgaggaacgc cgtgcgaacc ctttaccgag aggtggagcg agtagtatcc cgtccgtaga
      781 atctggtgct tccctgaact gcgtgaaata gtcaacagcc gacaataact gcggccgcac
      841 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
      901 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
      961 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     1021 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1081 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     1141 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1201 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1261 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1321 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     1381 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     1441 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     1501 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     1561 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gacaggtggc acttttcggg
     1621 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1681 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1741 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1801 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1861 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1921 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     1981 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2041 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2101 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     2161 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     2221 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     2281 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     2341 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     2401 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tcccgcggta
     2461 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     2521 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     2581 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     2641 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatcgc ggccgctgga
     2701 ctgttgtgat atgccatata tgcagtggtg ttcgccggcg ctggttccgg ctgtcttgct
     2761 tagttgtgag tcgccaggca gtcatcggta tgatctgtac atgattcgtc agtgtgagca
     2821 ccactgacga gcagatttcg ctgacatgaa tatgctggtc cctgcagtcc tactagtgaa
     2881 gcgtctcatg ctcagctgag acctataaac gcagaaaggc ccacccgaag gtgagccagt
     2941 gtgactctag tagagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttt
     3001 cgactgagcc tttcgtttta tttgatgcct ggtcatttgt acagttcatc cataccatgc
     3061 gtgatgcccg ctgcggttac gaactccagc agaaccatat gatcgcgttt ctcgttcgga
     3121 tctttagaca gaacgctttg cgtgctcaga tagtgattgt ctggcagcag aacaggacca
     3181 tcaccgattg gagtgttttg ctggtagtga tcagccagct gcacgctgcc atcctccacg
     3241 ttgtggcgaa ttttaaaatt cgctttaatg ccattttttt gtttatcggc gg
//
