LOCUS       sequence-369412-        3292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Assembly cassette with ConL2 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: pMYT042
FEATURES             Location/Qualifiers
     source          1..3292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             455..466
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     rep_origin      complement(929..1517)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1009..1028)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1620..1724
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1725..1793,1794..2585)
                     /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(1943..1962)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      complement(2901..2928)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2944..3015)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(3027..3292,1..448))
                     /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 cattgtggct gttaaaattg tattccagct tatggcccag gatattgccg tcctctttaa
       61 agtcaatgcc tttcagctca atgcggttta ccagggtatc gccttcaaat ttcacttccg
      121 cacgcgtttt gtacgtgccg tcatccttaa aggaaatcgt gcgttcctgc acatagcctt
      181 ccggcatggc ggacttgaag aagtcatgct gcttcatatg gtccggataa cgagcaaagc
      241 actgaacacc ataagtcagc gtcgttacca gagtcggcca aggaaccggc agtttaccag
      301 tagtacagat gaacttcagc gtcagtttac cattagttgc gtcaccttca ccctcgccac
      361 gcacggaaaa cttatgaccg ttgacatcac catccagttc caccagaata gggacgacac
      421 cagtgaacag ttcttcgcct ttacgcatct agtatttctc ctctttctct agtagctaag
      481 atccgaacct gccgtttctt gagttgccgc gatgttaaga aaacattcat aaattaaatg
      541 tgaattgccg cacacattat taaataagat ttacaaaatg ttcaaaatga cgcatgaaat
      601 cacgtttcac tttcggtctc acgttcatct gagacgcgat tctagagact gaattcgagc
      661 tgggagattg tgtctggcgg ctcacgtctt atttgggcaa aggagtcagc ggatcaccgt
      721 actatgtgtg agtgctccta tcgcgcttgc tcctgtgaga tacgatgtgg ccgtgtctga
      781 ccgtggtgcg tcatctattc actcacttgg ctgcgattta tgcggccgca cggttatcca
      841 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
      901 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
      961 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1021 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1081 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1141 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1201 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1261 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1321 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1381 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1441 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1501 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1561 acgaaaactc acgttaaggg attttggtca tgacaggtgg cacttttcgg ggaaatgtgc
     1621 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1681 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     1741 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     1801 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     1861 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     1921 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     1981 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2041 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2101 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2161 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2221 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     2281 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2341 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2401 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtcccgcggt atcattgcag
     2461 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2521 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2581 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     2641 aatttaaaag gatctaggtg aagatccttt ttgataatcg cggccgctgg actgttgtga
     2701 tatgccatat atgcagtggt gttcgccggc gctggttccg gctgtcttgc ttagttgtga
     2761 gtcgccaggc agtcatcggt atgatctgta catgattcgt cagtgtgagc accactgacg
     2821 agcagatttc gctgacatga atatgctggt ccctgcagac tgactagtgc atcgtctcat
     2881 gctcagctga gacctataaa cgcagaaagg cccacccgaa ggtgagccag tgtgactcta
     2941 gtagagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt tcgactgagc
     3001 ctttcgtttt atttgatgcc tggtcatttg tacagttcat ccataccatg cgtgatgccc
     3061 gctgcggtta cgaactccag cagaaccata tgatcgcgtt tctcgttcgg atctttagac
     3121 agaacgcttt gcgtgctcag atagtgattg tctggcagca gaacaggacc atcaccgatt
     3181 ggagtgtttt gctggtagtg atcagccagc tgcacgctgc catcctccac gttgtggcga
     3241 attttaaaat tcgctttaat gccatttttt tgtttatcgg cggtgatgta aa
//
