LOCUS       sequence-369605-        3292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Assembly cassette with ConL5 and ConR6 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: pMYT049
FEATURES             Location/Qualifiers
     source          1..3292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      635..706
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      722..749
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(1065..1856,1857..1925))
                     /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     1688..1707
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1926..2030)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2133..2721
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2622..2641
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     RBS             3184..3195
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(3202..3292,1..623)
                     /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 gcgagggtga aggtgacgca actaatggta aactgacgct gaagttcatc tgtactactg
       61 gtaaactgcc ggttccttgg ccgactctgg taacgacgct gacttatggt gttcagtgct
      121 ttgctcgtta tccggaccat atgaagcagc atgacttctt caagtccgcc atgccggaag
      181 gctatgtgca ggaacgcacg atttccttta aggatgacgg cacgtacaaa acgcgtgcgg
      241 aagtgaaatt tgaaggcgat accctggtaa accgcattga gctgaaaggc attgacttta
      301 aagaggacgg caatatcctg ggccataagc tggaatacaa ttttaacagc cacaatgttt
      361 acatcaccgc cgataaacaa aaaaatggca ttaaagcgaa ttttaaaatt cgccacaacg
      421 tggaggatgg cagcgtgcag ctggctgatc actaccagca aaacactcca atcggtgatg
      481 gtcctgttct gctgccagac aatcactatc tgagcacgca aagcgttctg tctaaagatc
      541 cgaacgagaa acgcgatcat atggttctgc tggagttcgt aaccgcagcg ggcatcacgc
      601 atggtatgga tgaactgtac aaatgaccag gcatcaaata aaacgaaagg ctcagtcgaa
      661 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact agagtcacac
      721 tggctcacct tcgggtgggc ctttctgcgt ttataggtct cagctgtaga tgagacgtcc
      781 tactagtcac actgcaggca gacacagtaa gacacgggcc ctgcggtaaa cgaaagttat
      841 ctgctctggc gtaagtctcc tactgtgtct gagtcaccct gattattcac gaggctattg
      901 taaagcgaag ttgggaaggt atcggtctac ggtaacggaa ccactatctc ggagcacacg
      961 acgcggccgc gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     1021 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     1081 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     1141 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     1201 ccgcgggacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     1261 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     1321 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     1381 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     1441 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     1501 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     1561 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     1621 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     1681 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     1741 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     1801 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     1861 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     1921 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     1981 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     2041 cgaaaagtgc cacctgtcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     2101 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2161 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2221 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2281 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2341 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2401 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2461 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2521 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2581 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2641 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2701 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     2761 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     2821 gcggccgcac gcacaaggtc agggcactca tgcgacaatc aactcgatgc atgatccgca
     2881 ccattgtcga ggggccagcg tcaatagtgc cgatgaccac agacccggtt aagacatagc
     2941 cgaatggagc cgcgccgacc acagaatgat agcggtgtag cgaaggatca agggaattca
     3001 gagtctagat ggtcgtctca aagtaacgtg agaccgaaag tgaaacgtga tttcatgcgt
     3061 cattttgaac attttgtaaa tcttatttaa taatgtgtgc ggcaattcac atttaattta
     3121 tgaatgtttt cttaacatcg cggcaactca agaaacggca ggttcggatc ttagctacta
     3181 gagaaagagg agaaatacta gatgcgtaaa ggcgaagaac tgttcactgg tgtcgtccct
     3241 attctggtgg aactggatgg tgatgtcaac ggtcataagt tttccgtgcg tg
//
