LOCUS       sequence-369607-        3292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Assembly cassette with ConL6 and ConR7 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: pMYT051
FEATURES             Location/Qualifiers
     source          1..3292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             17..28
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             35..748
                     /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"
     terminator      760..831
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      847..874
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(1190..1981,1982..2050))
                     /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     1813..1832
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2051..2155)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2258..2846
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2747..2766
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atcttagcta ctagagaaag aggagaaata ctagatgcgt aaaggcgaag aactgttcac
       61 tggtgtcgtc cctattctgg tggaactgga tggtgatgtc aacggtcata agttttccgt
      121 gcgtggcgag ggtgaaggtg acgcaactaa tggtaaactg acgctgaagt tcatctgtac
      181 tactggtaaa ctgccggttc cttggccgac tctggtaacg acgctgactt atggtgttca
      241 gtgctttgct cgttatccgg accatatgaa gcagcatgac ttcttcaagt ccgccatgcc
      301 ggaaggctat gtgcaggaac gcacgatttc ctttaaggat gacggcacgt acaaaacgcg
      361 tgcggaagtg aaatttgaag gcgataccct ggtaaaccgc attgagctga aaggcattga
      421 ctttaaagag gacggcaata tcctgggcca taagctggaa tacaatttta acagccacaa
      481 tgtttacatc accgccgata aacaaaaaaa tggcattaaa gcgaatttta aaattcgcca
      541 caacgtggag gatggcagcg tgcagctggc tgatcactac cagcaaaaca ctccaatcgg
      601 tgatggtcct gttctgctgc cagacaatca ctatctgagc acgcaaagcg ttctgtctaa
      661 agatccgaac gagaaacgcg atcatatggt tctgctggag ttcgtaaccg cagcgggcat
      721 cacgcatggt atggatgaac tgtacaaatg accaggcatc aaataaaacg aaaggctcag
      781 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt
      841 cacactggct caccttcggg tgggcctttc tgcgtttata ggtctcagct gacgatgaga
      901 cggtgtacta gtggaactgc agcggtcgga ctattcattg tggcagttat tgtcggctgt
      961 tgactatttc acgcagttca gggaagcacc agattctacg gacgggatac tactcgctcc
     1021 acctctcggt aaagggttcg cacggcgttc ctcacagata cgattttcgg gtctacagtt
     1081 gacgagcgcg gccgcgatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     1141 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     1201 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     1261 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     1321 tgataccgcg ggacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     1381 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     1441 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     1501 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     1561 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     1621 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     1681 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     1741 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     1801 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     1861 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     1921 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     1981 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     2041 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     2101 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     2161 ttccccgaaa agtgccacct gtcatgacca aaatccctta acgtgagttt tcgttccact
     2221 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     2281 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     2341 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2401 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     2461 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     2521 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2581 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2641 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2701 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2761 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2821 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2881 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2941 accgtgcggc cgccctgcgg taaacgaaag ttatctgctc tggcgtaagt ctcctactgt
     3001 gtctgagtca ccctgattat tcacgaggct attgtaaagc gaagttggga aggtatcggt
     3061 ctacggtaac ggaaccacta tctcggagca cacgacgcag aaggacgagt caccatgtga
     3121 attctccttc tagacacacg tctcatagaa acgtgagacc gaaagtgaaa cgtgatttca
     3181 tgcgtcattt tgaacatttt gtaaatctta tttaataatg tgtgcggcaa ttcacattta
     3241 atttatgaat gttttcttaa catcgcggca actcaagaaa cggcaggttc gg
//
