LOCUS       sequence-369606-        3292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Assembly cassette with ConL6 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: pMYT050
FEATURES             Location/Qualifiers
     source          1..3292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             415..426
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     rep_origin      complement(889..1477)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(969..988)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1580..1684
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1685..1753,1754..2545)
                     /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(1903..1922)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      complement(2861..2888)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2904..2975)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(2987..3292,1..408))
                     /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 gatattgccg tcctctttaa agtcaatgcc tttcagctca atgcggttta ccagggtatc
       61 gccttcaaat ttcacttccg cacgcgtttt gtacgtgccg tcatccttaa aggaaatcgt
      121 gcgttcctgc acatagcctt ccggcatggc ggacttgaag aagtcatgct gcttcatatg
      181 gtccggataa cgagcaaagc actgaacacc ataagtcagc gtcgttacca gagtcggcca
      241 aggaaccggc agtttaccag tagtacagat gaacttcagc gtcagtttac cattagttgc
      301 gtcaccttca ccctcgccac gcacggaaaa cttatgaccg ttgacatcac catccagttc
      361 caccagaata gggacgacac cagtgaacag ttcttcgcct ttacgcatct agtatttctc
      421 ctctttctct agtagctaag atccgaacct gccgtttctt gagttgccgc gatgttaaga
      481 aaacattcat aaattaaatg tgaattgccg cacacattat taaataagat ttacaaaatg
      541 ttcaaaatga cgcatgaaat cacgtttcac tttcggtctc acgtttctat gagacgtggt
      601 tctagaacca gaattcacat ggtgactcgt ccttctgcgt cgtgtgctcc gagatagtgg
      661 ttccgttacc gtagaccgat accttcccaa cttcgcttta caatagcctc gtgaataatc
      721 agggtgactc agacacagta ggagacttac gccagagcag ataactttcg tttaccgcag
      781 ggcggccgca cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
      841 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
      901 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
      961 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1021 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1081 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1141 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1201 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1261 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1321 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1381 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1441 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1501 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgacaggtgg
     1561 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     1621 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     1681 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     1741 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     1801 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     1861 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     1921 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     1981 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     2041 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     2101 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     2161 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     2221 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     2281 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     2341 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     2401 gtcccgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     2461 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     2521 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     2581 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatcg
     2641 cggccgctgg actgttgtga tatgccatat atgcagtggt gttcgccggc gctggttccg
     2701 gctgtcttgc ttagttgtga gtcgccaggc agtcatcggt atgatctgta catgattcgt
     2761 cagtgtgagc accactgacg agcagatttc gctgacatga atatgctggt ccctgcagac
     2821 acactagttg tgcgtctcat gctcagctga gacctataaa cgcagaaagg cccacccgaa
     2881 ggtgagccag tgtgactcta gtagagagcg ttcaccgaca aacaacagat aaaacgaaag
     2941 gcccagtctt tcgactgagc ctttcgtttt atttgatgcc tggtcatttg tacagttcat
     3001 ccataccatg cgtgatgccc gctgcggtta cgaactccag cagaaccata tgatcgcgtt
     3061 tctcgttcgg atctttagac agaacgcttt gcgtgctcag atagtgattg tctggcagca
     3121 gaacaggacc atcaccgatt ggagtgtttt gctggtagtg atcagccagc tgcacgctgc
     3181 catcctccac gttgtggcga attttaaaat tcgctttaat gccatttttt tgtttatcgg
     3241 cggtgatgta aacattgtgg ctgttaaaat tgtattccag cttatggccc ag
//
