LOCUS       sequence-369416-        3292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Assembly cassette with ConL4 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: pMYT046
FEATURES             Location/Qualifiers
     source          1..3292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(62..650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(142..161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        753..857
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(858..926,927..1718)
                     /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(1076..1095)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      complement(2034..2061)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2077..2148)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2160..2873)
                     /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             2880..2891
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
       61 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      121 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      181 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      241 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      301 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      361 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      421 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      481 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      541 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      601 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      661 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      721 tcatgacagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
      781 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
      841 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
      901 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
      961 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     1021 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     1081 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     1141 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     1201 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     1261 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     1321 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     1381 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     1441 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     1501 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     1561 ccggtgagcg tgggtcccgc ggtatcattg cagcactggg gccagatggt aagccctccc
     1621 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     1681 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     1741 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     1801 tttttgataa tcgcggccgc tggactgttg tgatatgcca tatatgcagt ggtgttcgcc
     1861 ggcgctggtt ccggctgtct tgcttagttg tgagtcgcca ggcagtcatc ggtatgatct
     1921 gtacatgatt cgtcagtgtg agcaccactg acgagcagat ttcgctgaca tgaatatgct
     1981 ggtccctgca gctagactag ttacgcgtct catgctcagc tgagacctat aaacgcagaa
     2041 aggcccaccc gaaggtgagc cagtgtgact ctagtagaga gcgttcaccg acaaacaaca
     2101 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctggtcat
     2161 ttgtacagtt catccatacc atgcgtgatg cccgctgcgg ttacgaactc cagcagaacc
     2221 atatgatcgc gtttctcgtt cggatcttta gacagaacgc tttgcgtgct cagatagtga
     2281 ttgtctggca gcagaacagg accatcaccg attggagtgt tttgctggta gtgatcagcc
     2341 agctgcacgc tgccatcctc cacgttgtgg cgaattttaa aattcgcttt aatgccattt
     2401 ttttgtttat cggcggtgat gtaaacattg tggctgttaa aattgtattc cagcttatgg
     2461 cccaggatat tgccgtcctc tttaaagtca atgcctttca gctcaatgcg gtttaccagg
     2521 gtatcgcctt caaatttcac ttccgcacgc gttttgtacg tgccgtcatc cttaaaggaa
     2581 atcgtgcgtt cctgcacata gccttccggc atggcggact tgaagaagtc atgctgcttc
     2641 atatggtccg gataacgagc aaagcactga acaccataag tcagcgtcgt taccagagtc
     2701 ggccaaggaa ccggcagttt accagtagta cagatgaact tcagcgtcag tttaccatta
     2761 gttgcgtcac cttcaccctc gccacgcacg gaaaacttat gaccgttgac atcaccatcc
     2821 agttccacca gaatagggac gacaccagtg aacagttctt cgcctttacg catctagtat
     2881 ttctcctctt tctctagtag ctaagatccg aacctgccgt ttcttgagtt gccgcgatgt
     2941 taagaaaaca ttcataaatt aaatgtgaat tgccgcacac attattaaat aagatttaca
     3001 aaatgttcaa aatgacgcat gaaatcacgt ttcactttcg gtctcacgtt tacctgagac
     3061 gggaatctag acatggaatt cggaatacga gacaccaagc agcgtcaggc tgtctctttg
     3121 cattgtcacg gtttcggtta gcctgcctcg atttcaggat gtggctcggt gttacgtcat
     3181 ctgtccagcg ttgtcaccat gtttagacgg attggacgca tcttgcctag ttcgtctagt
     3241 gcaggcgcgg ccgcacggtt atccacagaa tcaggggata acgcaggaaa ga
//
