LOCUS       sequence-369604-        3292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Assembly cassette with ConL5 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: pMYT048
FEATURES             Location/Qualifiers
     source          1..3292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(305..893)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(385..404)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        996..1100
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1101..1169,1170..1961)
                     /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(1319..1338)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      complement(2277..2304)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2320..2391)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2403..3116)
                     /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             3123..3134
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 tactttgaga cgtcagtcta gactaggaat tcccttgatc cttcgctaca ccgctatcat
       61 tctgtggtcg gcgcggctcc attcggctat gtcttaaccg ggtctgtggt catcggcact
      121 attgacgctg gcccctcgac aatggtgcgg atcatgcatc gagttgattg tcgcatgagt
      181 gccctgacct tgtgcgtgcg gccgcacggt tatccacaga atcaggggat aacgcaggaa
      241 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      301 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      361 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      421 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      481 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
      541 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
      601 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
      661 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
      721 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
      781 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
      841 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
      901 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
      961 tggtcatgac aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1021 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1081 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1141 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1201 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     1261 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     1321 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     1381 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     1441 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     1501 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     1561 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     1621 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     1681 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     1741 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     1801 gagccggtga gcgtgggtcc cgcggtatca ttgcagcact ggggccagat ggtaagccct
     1861 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1921 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1981 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2041 tcctttttga taatcgcggc cgctggactg ttgtgatatg ccatatatgc agtggtgttc
     2101 gccggcgctg gttccggctg tcttgcttag ttgtgagtcg ccaggcagtc atcggtatga
     2161 tctgtacatg attcgtcagt gtgagcacca ctgacgagca gatttcgctg acatgaatat
     2221 gctggtccct gcagggttac tagtagtccg tctcatgctc agctgagacc tataaacgca
     2281 gaaaggccca cccgaaggtg agccagtgtg actctagtag agagcgttca ccgacaaaca
     2341 acagataaaa cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgcctggt
     2401 catttgtaca gttcatccat accatgcgtg atgcccgctg cggttacgaa ctccagcaga
     2461 accatatgat cgcgtttctc gttcggatct ttagacagaa cgctttgcgt gctcagatag
     2521 tgattgtctg gcagcagaac aggaccatca ccgattggag tgttttgctg gtagtgatca
     2581 gccagctgca cgctgccatc ctccacgttg tggcgaattt taaaattcgc tttaatgcca
     2641 tttttttgtt tatcggcggt gatgtaaaca ttgtggctgt taaaattgta ttccagctta
     2701 tggcccagga tattgccgtc ctctttaaag tcaatgcctt tcagctcaat gcggtttacc
     2761 agggtatcgc cttcaaattt cacttccgca cgcgttttgt acgtgccgtc atccttaaag
     2821 gaaatcgtgc gttcctgcac atagccttcc ggcatggcgg acttgaagaa gtcatgctgc
     2881 ttcatatggt ccggataacg agcaaagcac tgaacaccat aagtcagcgt cgttaccaga
     2941 gtcggccaag gaaccggcag tttaccagta gtacagatga acttcagcgt cagtttacca
     3001 ttagttgcgt caccttcacc ctcgccacgc acggaaaact tatgaccgtt gacatcacca
     3061 tccagttcca ccagaatagg gacgacacca gtgaacagtt cttcgccttt acgcatctag
     3121 tatttctcct ctttctctag tagctaagat ccgaacctgc cgtttcttga gttgccgcga
     3181 tgttaagaaa acattcataa attaaatgtg aattgccgca cacattatta aataagattt
     3241 acaaaatgtt caaaatgacg catgaaatca cgtttcactt tcggtctcac gt
//
