LOCUS       sequence-369382-        3292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Assembly cassette with ConLS and ConR1 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: pMYT039
FEATURES             Location/Qualifiers
     source          1..3292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             82..93
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     rep_origin      complement(556..1144)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(636..655)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1247..1351
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1352..1420,1421..2212)
                     /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(1570..1589)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      complement(2528..2555)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2571..2642)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(2654..3292,1..75))
                     /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 atgaccgttg acatcaccat ccagttccac cagaataggg acgacaccag tgaacagttc
       61 ttcgccttta cgcatctagt atttctcctc tttctctagt agctaagatc cgaacctgcc
      121 gtttcttgag ttgccgcgat gttaagaaaa cattcataaa ttaaatgtga attgccgcac
      181 acattattaa ataagattta caaaatgttc aaaatgacgc atgaaatcac gtttcacttt
      241 cggtctcacg tttcagtgag acgcagttct agaacgtgaa ttcgacctag ccacatttcc
      301 aaggcgtatt gcgacgaatt gccacgttgt cgcagtgtct gtactgtggg ttaatgtctg
      361 tgatcgccgt cgtgagttcg tgtcatccgc gcatgattca gatggtcctg gagatcgttg
      421 cttgtaccgg ctgtttgtgg ctctaccagc ggccgcacgg ttatccacag aatcagggga
      481 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      541 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      601 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      661 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      721 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      781 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      841 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      901 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      961 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1021 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1081 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1141 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1201 ttaagggatt ttggtcatga caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     1261 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     1321 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     1381 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     1441 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     1501 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     1561 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     1621 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     1681 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     1741 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     1801 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     1861 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     1921 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     1981 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     2041 tgataaatct ggagccggtg agcgtgggtc ccgcggtatc attgcagcac tggggccaga
     2101 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     2161 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     2221 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     2281 ctaggtgaag atcctttttg ataatcgcgg ccgctaactg ccttgatctg tcgggtgtct
     2341 gcttctctca caggcttagg tggatctgct gtccccagtc agtgcggtca tcatgcagtc
     2401 atccgagcgt gtattgcaca catttgtttg gcgcatgatg agccgtgatg accccgtgca
     2461 gcaaacgagg accgattagc tgcaggacta ctagtagctc gtctcattgg cagctgagac
     2521 ctataaacgc agaaaggccc acccgaaggt gagccagtgt gactctagta gagagcgttc
     2581 accgacaaac aacagataaa acgaaaggcc cagtctttcg actgagcctt tcgttttatt
     2641 tgatgcctgg tcatttgtac agttcatcca taccatgcgt gatgcccgct gcggttacga
     2701 actccagcag aaccatatga tcgcgtttct cgttcggatc tttagacaga acgctttgcg
     2761 tgctcagata gtgattgtct ggcagcagaa caggaccatc accgattgga gtgttttgct
     2821 ggtagtgatc agccagctgc acgctgccat cctccacgtt gtggcgaatt ttaaaattcg
     2881 ctttaatgcc atttttttgt ttatcggcgg tgatgtaaac attgtggctg ttaaaattgt
     2941 attccagctt atggcccagg atattgccgt cctctttaaa gtcaatgcct ttcagctcaa
     3001 tgcggtttac cagggtatcg ccttcaaatt tcacttccgc acgcgttttg tacgtgccgt
     3061 catccttaaa ggaaatcgtg cgttcctgca catagccttc cggcatggcg gacttgaaga
     3121 agtcatgctg cttcatatgg tccggataac gagcaaagca ctgaacacca taagtcagcg
     3181 tcgttaccag agtcggccaa ggaaccggca gtttaccagt agtacagatg aacttcagcg
     3241 tcagtttacc attagttgcg tcaccttcac cctcgccacg cacggaaaac tt
//
