LOCUS       sequence-369610-        3292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Assembly cassette with ConL8 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: pMYT054
FEATURES             Location/Qualifiers
     source          1..3292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             466..477
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     rep_origin      complement(940..1528)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1020..1039)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1631..1735
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1736..1804,1805..2596)
                     /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(1954..1973)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      complement(2912..2939)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2955..3026)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(3038..3292,1..459))
                     /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 ggtgatgtaa acattgtggc tgttaaaatt gtattccagc ttatggccca ggatattgcc
       61 gtcctcttta aagtcaatgc ctttcagctc aatgcggttt accagggtat cgccttcaaa
      121 tttcacttcc gcacgcgttt tgtacgtgcc gtcatcctta aaggaaatcg tgcgttcctg
      181 cacatagcct tccggcatgg cggacttgaa gaagtcatgc tgcttcatat ggtccggata
      241 acgagcaaag cactgaacac cataagtcag cgtcgttacc agagtcggcc aaggaaccgg
      301 cagtttacca gtagtacaga tgaacttcag cgtcagttta ccattagttg cgtcaccttc
      361 accctcgcca cgcacggaaa acttatgacc gttgacatca ccatccagtt ccaccagaat
      421 agggacgaca ccagtgaaca gttcttcgcc tttacgcatc tagtatttct cctctttctc
      481 tagtagctaa gatccgaacc tgccgtttct tgagttgccg cgatgttaag aaaacattca
      541 taaattaaat gtgaattgcc gcacacatta ttaaataaga tttacaaaat gttcaaaatg
      601 acgcatgaaa tcacgtttca ctttcggtct cacgttggag tgagacgtcc ttctagagaa
      661 ggaattcgtc cgtcattgca tctaggtgcc taccaacctc agtaatacga aacggattga
      721 cctgctcctc tcgccgaaga agtccctgga atagtaaaaa ctgcccaacg cacgattgcc
      781 ttatctgtag acacggtcct gtgactgaac tctcccgacg aaatagaata cggcggccgc
      841 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
      901 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
      961 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1021 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1081 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1141 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1201 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1261 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1321 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1381 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1441 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1501 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1561 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgacaggtg gcacttttcg
     1621 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1681 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1741 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1801 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     1861 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     1921 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     1981 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2041 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2101 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     2161 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2221 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     2281 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2341 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2401 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtcccgcgg
     2461 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2521 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2581 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2641 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc gcggccgctg
     2701 gactgttgtg atatgccata tatgcagtgg tgttcgccgg cgctggttcc ggctgtcttg
     2761 cttagttgtg agtcgccagg cagtcatcgg tatgatctgt acatgattcg tcagtgtgag
     2821 caccactgac gagcagattt cgctgacatg aatatgctgg tccctgcagc aacactagtg
     2881 tgtcgtctca tgctcagctg agacctataa acgcagaaag gcccacccga aggtgagcca
     2941 gtgtgactct agtagagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
     3001 ttcgactgag cctttcgttt tatttgatgc ctggtcattt gtacagttca tccataccat
     3061 gcgtgatgcc cgctgcggtt acgaactcca gcagaaccat atgatcgcgt ttctcgttcg
     3121 gatctttaga cagaacgctt tgcgtgctca gatagtgatt gtctggcagc agaacaggac
     3181 catcaccgat tggagtgttt tgctggtagt gatcagccag ctgcacgctg ccatcctcca
     3241 cgttgtggcg aattttaaaa ttcgctttaa tgccattttt ttgtttatcg gc
//
