LOCUS       sequence-369603-        3292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Assembly cassette with ConL4 and ConR5 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: pMYT047
FEATURES             Location/Qualifiers
     source          1..3292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(335..362)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(378..449)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(461..1174)
                     /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"
     RBS             1181..1192
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     rep_origin      complement(1655..2243)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1735..1754)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2346..2450
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2451..2519,2520..3292,1..19)
                     /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(2669..2688)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
       61 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atcgcggccg
      121 ctatcattct gtggtcggcg cggctccatt cggctatgtc ttaaccgggt ctgtggtcat
      181 cggcactatt gacgctggcc cctcgacaat ggtgcggatc atgcatcgag ttgattgtcg
      241 catgagtgcc ctgaccttgt gcgtgctcgt gaaacatttc gtccatctgc agtcagacta
      301 gtctagcgtc tcaacttcag ctgagaccta taaacgcaga aaggcccacc cgaaggtgag
      361 ccagtgtgac tctagtagag agcgttcacc gacaaacaac agataaaacg aaaggcccag
      421 tctttcgact gagcctttcg ttttatttga tgcctggtca tttgtacagt tcatccatac
      481 catgcgtgat gcccgctgcg gttacgaact ccagcagaac catatgatcg cgtttctcgt
      541 tcggatcttt agacagaacg ctttgcgtgc tcagatagtg attgtctggc agcagaacag
      601 gaccatcacc gattggagtg ttttgctggt agtgatcagc cagctgcacg ctgccatcct
      661 ccacgttgtg gcgaatttta aaattcgctt taatgccatt tttttgttta tcggcggtga
      721 tgtaaacatt gtggctgtta aaattgtatt ccagcttatg gcccaggata ttgccgtcct
      781 ctttaaagtc aatgcctttc agctcaatgc ggtttaccag ggtatcgcct tcaaatttca
      841 cttccgcacg cgttttgtac gtgccgtcat ccttaaagga aatcgtgcgt tcctgcacat
      901 agccttccgg catggcggac ttgaagaagt catgctgctt catatggtcc ggataacgag
      961 caaagcactg aacaccataa gtcagcgtcg ttaccagagt cggccaagga accggcagtt
     1021 taccagtagt acagatgaac ttcagcgtca gtttaccatt agttgcgtca ccttcaccct
     1081 cgccacgcac ggaaaactta tgaccgttga catcaccatc cagttccacc agaataggga
     1141 cgacaccagt gaacagttct tcgcctttac gcatctagta tttctcctct ttctctagta
     1201 gctaagatcc gaacctgccg tttcttgagt tgccgcgatg ttaagaaaac attcataaat
     1261 taaatgtgaa ttgccgcaca cattattaaa taagatttac aaaatgttca aaatgacgca
     1321 tgaaatcacg tttcactttc ggtctcacgt ttacctgaga cgtacgtcta gaatcggaat
     1381 tcggaatacg agacaccaag cagcgtcagg ctgtctcttt gcattgtcac ggtttcggtt
     1441 agcctgcctc gatttcagga tgtggctcgg tgttacgtca tctgtccagc gttgtcacca
     1501 tgtttagacg gattggacgc atcttgccta gttcgtctag tgcaggcgcg gccgcacggt
     1561 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     1621 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1681 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1741 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1801 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1861 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1921 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1981 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2041 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2101 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2161 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2221 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2281 agtggaacga aaactcacgt taagggattt tggtcatgac aggtggcact tttcggggaa
     2341 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2401 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2461 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2521 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2581 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2641 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     2701 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2761 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2821 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2881 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2941 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3001 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3061 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3121 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtcc cgcggtatca
     3181 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3241 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tc
//
