LOCUS       sequence-369609-        3292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Assembly cassette with ConL7 and ConR8 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: pMYT053
FEATURES             Location/Qualifiers
     source          1..3292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             462..473
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     rep_origin      complement(936..1524)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1016..1035)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1627..1731
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1732..1800,1801..2592)
                     /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(1950..1969)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      complement(2908..2935)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2951..3022)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(3034..3292,1..455))
                     /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 atgtaaacat tgtggctgtt aaaattgtat tccagcttat ggcccaggat attgccgtcc
       61 tctttaaagt caatgccttt cagctcaatg cggtttacca gggtatcgcc ttcaaatttc
      121 acttccgcac gcgttttgta cgtgccgtca tccttaaagg aaatcgtgcg ttcctgcaca
      181 tagccttccg gcatggcgga cttgaagaag tcatgctgct tcatatggtc cggataacga
      241 gcaaagcact gaacaccata agtcagcgtc gttaccagag tcggccaagg aaccggcagt
      301 ttaccagtag tacagatgaa cttcagcgtc agtttaccat tagttgcgtc accttcaccc
      361 tcgccacgca cggaaaactt atgaccgttg acatcaccat ccagttccac cagaataggg
      421 acgacaccag tgaacagttc ttcgccttta cgcatctagt atttctcctc tttctctagt
      481 agctaagatc cgaacctgcc gtttcttgag ttgccgcgat gttaagaaaa cattcataaa
      541 ttaaatgtga attgccgcac acattattaa ataagattta caaaatgttc aaaatgacgc
      601 atgaaatcac gtttcacttt cggtctcacg tttcgttgag acggaagtct agaacacgaa
      661 ttcagcacct ttcccacgta aacgcgctcg tcaactgtag acccgaaaat cgtatctgtg
      721 aggaacgccg tgcgaaccct ttaccgagag gtggagcgag tagtatcccg tccgtagaat
      781 ctggtgcttc cctgaactgc gtgaaatagt caacagccga caataactgc ggccgcacgg
      841 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
      901 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
      961 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1021 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     1081 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     1141 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     1201 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     1261 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     1321 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     1381 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     1441 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     1501 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     1561 cagtggaacg aaaactcacg ttaagggatt ttggtcatga caggtggcac ttttcgggga
     1621 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1681 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     1741 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     1801 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     1861 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     1921 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     1981 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2041 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2101 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2161 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2221 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     2281 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2341 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2401 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc ccgcggtatc
     2461 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2521 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2581 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     2641 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatcgcgg ccgcctacca
     2701 acctcagtaa tacgaaacgg attgacctgc tcctctcgcc gaagaagtcc ctggaatagt
     2761 aaaaactgcc caacgcacga ttgccttatc tgtagacacg gtcctgtgac tgaactctcc
     2821 cgacgaaata gaatacggca tgtacgatcg gatggactgc tgcaggtgta ctagttcctc
     2881 gtctcaggag cagctgagac ctataaacgc agaaaggccc acccgaaggt gagccagtgt
     2941 gactctagta gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtctttcg
     3001 actgagcctt tcgttttatt tgatgcctgg tcatttgtac agttcatcca taccatgcgt
     3061 gatgcccgct gcggttacga actccagcag aaccatatga tcgcgtttct cgttcggatc
     3121 tttagacaga acgctttgcg tgctcagata gtgattgtct ggcagcagaa caggaccatc
     3181 accgattgga gtgttttgct ggtagtgatc agccagctgc acgctgccat cctccacgtt
     3241 gtggcgaatt ttaaaattcg ctttaatgcc atttttttgt ttatcggcgg tg
//
