LOCUS       sequence-369411-        3292 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Assembly cassette with ConL1 and ConR2 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 Apr 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: pMYT041
FEATURES             Location/Qualifiers
     source          1..3292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             461..472
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     rep_origin      complement(935..1523)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1015..1034)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1626..1730
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1731..1799,1800..2591)
                     /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(1949..1968)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      complement(2907..2934)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2950..3021)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(3033..3292,1..454))
                     /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 tgtaaacatt gtggctgtta aaattgtatt ccagcttatg gcccaggata ttgccgtcct
       61 ctttaaagtc aatgcctttc agctcaatgc ggtttaccag ggtatcgcct tcaaatttca
      121 cttccgcacg cgttttgtac gtgccgtcat ccttaaagga aatcgtgcgt tcctgcacat
      181 agccttccgg catggcggac ttgaagaagt catgctgctt catatggtcc ggataacgag
      241 caaagcactg aacaccataa gtcagcgtcg ttaccagagt cggccaagga accggcagtt
      301 taccagtagt acagatgaac ttcagcgtca gtttaccatt agttgcgtca ccttcaccct
      361 cgccacgcac ggaaaactta tgaccgttga catcaccatc cagttccacc agaataggga
      421 cgacaccagt gaacagttct tcgcctttac gcatctagta tttctcctct ttctctagta
      481 gctaagatcc gaacctgccg tttcttgagt tgccgcgatg ttaagaaaac attcataaat
      541 taaatgtgaa ttgccgcaca cattattaaa taagatttac aaaatgttca aaatgacgca
      601 tgaaatcacg tttcactttc ggtctcacgt tttggtgaga cggacttcta gaagctgaat
      661 tcttctgtag ttcatcgggt ccgctaactg ccttgatctg tcgggtgtct gcttctctca
      721 caggcttagg tggatctgct gtccccagtc agtgcggtca tcatgcagtc atccgagcgt
      781 gtattgcaca catttgtttg gcgcatgatg agccgtgatg accccgtgcg gccgcacggt
      841 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
      901 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      961 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1021 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1081 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1141 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1201 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1261 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1321 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1381 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1441 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1501 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1561 agtggaacga aaactcacgt taagggattt tggtcatgac aggtggcact tttcggggaa
     1621 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1681 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     1741 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     1801 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     1861 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     1921 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     1981 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2041 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2101 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2161 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2221 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     2281 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2341 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2401 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtcc cgcggtatca
     2461 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2521 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2581 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     2641 atttttaatt taaaaggatc taggtgaaga tcctttttga taatcgcggc cgcggctcac
     2701 gtcttatttg ggcaaaggag tcagcggatc accgtactat gtgtgagtgc tcctatcgcg
     2761 cttgctcctg tgagatacga tgtggccgtg tctgaccgtg gtgcgtcatc tattcactca
     2821 cttggctgcg atttatgcta catgccccta atcggttcct gcaggcatac tagtccttcg
     2881 tctcacatcc agctgagacc tataaacgca gaaaggccca cccgaaggtg agccagtgtg
     2941 actctagtag agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga
     3001 ctgagccttt cgttttattt gatgcctggt catttgtaca gttcatccat accatgcgtg
     3061 atgcccgctg cggttacgaa ctccagcaga accatatgat cgcgtttctc gttcggatct
     3121 ttagacagaa cgctttgcgt gctcagatag tgattgtctg gcagcagaac aggaccatca
     3181 ccgattggag tgttttgctg gtagtgatca gccagctgca cgctgccatc ctccacgttg
     3241 tggcgaattt taaaattcgc tttaatgcca tttttttgtt tatcggcggt ga
//
