LOCUS       sequence-369613-        3292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Assembly cassette with ConL9 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: pMYT056
FEATURES             Location/Qualifiers
     source          1..3292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             421..432
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     rep_origin      complement(895..1483)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(975..994)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1586..1690
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1691..1759,1760..2551)
                     /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(1909..1928)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      complement(2867..2894)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2910..2981)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(2993..3292,1..414))
                     /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 gcccaggata ttgccgtcct ctttaaagtc aatgcctttc agctcaatgc ggtttaccag
       61 ggtatcgcct tcaaatttca cttccgcacg cgttttgtac gtgccgtcat ccttaaagga
      121 aatcgtgcgt tcctgcacat agccttccgg catggcggac ttgaagaagt catgctgctt
      181 catatggtcc ggataacgag caaagcactg aacaccataa gtcagcgtcg ttaccagagt
      241 cggccaagga accggcagtt taccagtagt acagatgaac ttcagcgtca gtttaccatt
      301 agttgcgtca ccttcaccct cgccacgcac ggaaaactta tgaccgttga catcaccatc
      361 cagttccacc agaataggga cgacaccagt gaacagttct tcgcctttac gcatctagta
      421 tttctcctct ttctctagta gctaagatcc gaacctgccg tttcttgagt tgccgcgatg
      481 ttaagaaaac attcataaat taaatgtgaa ttgccgcaca cattattaaa taagatttac
      541 aaaatgttca aaatgacgca tgaaatcacg tttcactttc ggtctcacgt tcccttgaga
      601 cgcaactcta gagtgtgaat tcactgggca ggaagaaagt gtgcgcggaa tcaccgagca
      661 gtcctctact cctacaatcc acgagaacgg cgaatacagt tacgaacaga agtttacccg
      721 aaaagtcagc ctcccttact atcccagatt atcagtctat tgctccaacc tgcggacagc
      781 gtgtgctgcg gccgcacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
      841 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
      901 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
      961 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1021 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1081 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1141 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1201 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1261 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1321 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1381 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1441 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1501 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgac
     1561 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1621 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1681 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     1741 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     1801 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     1861 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     1921 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     1981 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2041 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     2101 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2161 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2221 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2281 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2341 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2401 gcgtgggtcc cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2461 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     2521 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     2581 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     2641 taatcgcggc cgctggactg ttgtgatatg ccatatatgc agtggtgttc gccggcgctg
     2701 gttccggctg tcttgcttag ttgtgagtcg ccaggcagtc atcggtatga tctgtacatg
     2761 attcgtcagt gtgagcacca ctgacgagca gatttcgctg acatgaatat gctggtccct
     2821 gcagagagac tagtcacacg tctcatgctc agctgagacc tataaacgca gaaaggccca
     2881 cccgaaggtg agccagtgtg actctagtag agagcgttca ccgacaaaca acagataaaa
     2941 cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgcctggt catttgtaca
     3001 gttcatccat accatgcgtg atgcccgctg cggttacgaa ctccagcaga accatatgat
     3061 cgcgtttctc gttcggatct ttagacagaa cgctttgcgt gctcagatag tgattgtctg
     3121 gcagcagaac aggaccatca ccgattggag tgttttgctg gtagtgatca gccagctgca
     3181 cgctgccatc ctccacgttg tggcgaattt taaaattcgc tttaatgcca tttttttgtt
     3241 tatcggcggt gatgtaaaca ttgtggctgt taaaattgta ttccagctta tg
//
