LOCUS       sequence-369612-        3292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Assembly cassette with ConL8 and ConR9 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: pMYT055
FEATURES             Location/Qualifiers
     source          1..3292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      631..702
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      718..745
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(1061..1852,1853..1921))
                     /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     1684..1703
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1922..2026)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2129..2717
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2618..2637
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     RBS             3180..3191
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(3198..3292,1..619)
                     /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 gggtgaaggt gacgcaacta atggtaaact gacgctgaag ttcatctgta ctactggtaa
       61 actgccggtt ccttggccga ctctggtaac gacgctgact tatggtgttc agtgctttgc
      121 tcgttatccg gaccatatga agcagcatga cttcttcaag tccgccatgc cggaaggcta
      181 tgtgcaggaa cgcacgattt cctttaagga tgacggcacg tacaaaacgc gtgcggaagt
      241 gaaatttgaa ggcgataccc tggtaaaccg cattgagctg aaaggcattg actttaaaga
      301 ggacggcaat atcctgggcc ataagctgga atacaatttt aacagccaca atgtttacat
      361 caccgccgat aaacaaaaaa atggcattaa agcgaatttt aaaattcgcc acaacgtgga
      421 ggatggcagc gtgcagctgg ctgatcacta ccagcaaaac actccaatcg gtgatggtcc
      481 tgttctgctg ccagacaatc actatctgag cacgcaaagc gttctgtcta aagatccgaa
      541 cgagaaacgc gatcatatgg ttctgctgga gttcgtaacc gcagcgggca tcacgcatgg
      601 tatggatgaa ctgtacaaat gaccaggcat caaataaaac gaaaggctca gtcgaaagac
      661 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc
      721 tcaccttcgg gtgggccttt ctgcgtttat aggtctcagc tgagggtgag acggttgact
      781 agttgtgctg cagtgactaa ctgtgccaaa tcggcagcac acgctgtccg caggttggag
      841 caatagactg ataatctggg atagtaaggg aggctgactt ttcgggtaaa cttctgttcg
      901 taactgtatt cgccgttctc gtggattgta ggagtagagg actgctcggt gattccgcgc
      961 ggccgcgatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1021 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1081 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1141 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1201 gggacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1261 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1321 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1381 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1441 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1501 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1561 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1621 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1681 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1741 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1801 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1861 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1921 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1981 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     2041 aagtgccacc tgtcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2101 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2161 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2221 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     2281 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2341 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2401 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2461 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2521 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2581 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2641 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2701 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     2761 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtgcgg
     2821 ccgccgtatt ctatttcgtc gggagagttc agtcacagga ccgtgtctac agataaggca
     2881 atcgtgcgtt gggcagtttt tactattcca gggacttctt cggcgagagg agcaggtcaa
     2941 tccgtttcgt attactgagg ttggtaggca cctagatgca atgacggacg aattccaact
     3001 ctagaacacc gtctcactcc aacgtgagac cgaaagtgaa acgtgatttc atgcgtcatt
     3061 ttgaacattt tgtaaatctt atttaataat gtgtgcggca attcacattt aatttatgaa
     3121 tgttttctta acatcgcggc aactcaagaa acggcaggtt cggatcttag ctactagaga
     3181 aagaggagaa atactagatg cgtaaaggcg aagaactgtt cactggtgtc gtccctattc
     3241 tggtggaact ggatggtgat gtcaacggtc ataagttttc cgtgcgtggc ga
//
