LOCUS       sequence-369414-        3292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Assembly cassette with ConL3 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: pMYT044
FEATURES             Location/Qualifiers
     source          1..3292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             463..474
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     rep_origin      complement(937..1525)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1017..1036)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1628..1732
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1733..1801,1802..2593)
                     /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(1951..1970)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      complement(2909..2936)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2952..3023)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(3035..3292,1..456))
                     /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 gatgtaaaca ttgtggctgt taaaattgta ttccagctta tggcccagga tattgccgtc
       61 ctctttaaag tcaatgcctt tcagctcaat gcggtttacc agggtatcgc cttcaaattt
      121 cacttccgca cgcgttttgt acgtgccgtc atccttaaag gaaatcgtgc gttcctgcac
      181 atagccttcc ggcatggcgg acttgaagaa gtcatgctgc ttcatatggt ccggataacg
      241 agcaaagcac tgaacaccat aagtcagcgt cgttaccaga gtcggccaag gaaccggcag
      301 tttaccagta gtacagatga acttcagcgt cagtttacca ttagttgcgt caccttcacc
      361 ctcgccacgc acggaaaact tatgaccgtt gacatcacca tccagttcca ccagaatagg
      421 gacgacacca gtgaacagtt cttcgccttt acgcatctag tatttctcct ctttctctag
      481 tagctaagat ccgaacctgc cgtttcttga gttgccgcga tgttaagaaa acattcataa
      541 attaaatgtg aattgccgca cacattatta aataagattt acaaaatgtt caaaatgacg
      601 catgaaatca cgtttcactt tcggtctcac gttgaactga gacgactgtc tagagcatga
      661 attcgtggct ttacctcgta ttgcgcgtac tgggttcgag ttccctgatc cgtatgtcga
      721 atggctacca ggctgacgac gatgatatga gcgtgcatcg gcatcttgcg gtgttgatta
      781 gctgccactg gtggctgtcc tacatcagcc tcatgtggtc taggtctgcg cggccgcacg
      841 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      901 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      961 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1021 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1081 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1141 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1201 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1261 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1321 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1381 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1441 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1501 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1561 tcagtggaac gaaaactcac gttaagggat tttggtcatg acaggtggca cttttcgggg
     1621 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     1681 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1741 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1801 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1861 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1921 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     1981 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     2041 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     2101 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     2161 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     2221 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     2281 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     2341 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     2401 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt cccgcggtat
     2461 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     2521 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     2581 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     2641 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatcgcg gccgctggac
     2701 tgttgtgata tgccatatat gcagtggtgt tcgccggcgc tggttccggc tgtcttgctt
     2761 agttgtgagt cgccaggcag tcatcggtat gatctgtaca tgattcgtca gtgtgagcac
     2821 cactgacgag cagatttcgc tgacatgaat atgctggtcc ctgcagaagg actagtcatg
     2881 cgtctcatgc tcagctgaga cctataaacg cagaaaggcc cacccgaagg tgagccagtg
     2941 tgactctagt agagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc
     3001 gactgagcct ttcgttttat ttgatgcctg gtcatttgta cagttcatcc ataccatgcg
     3061 tgatgcccgc tgcggttacg aactccagca gaaccatatg atcgcgtttc tcgttcggat
     3121 ctttagacag aacgctttgc gtgctcagat agtgattgtc tggcagcaga acaggaccat
     3181 caccgattgg agtgttttgc tggtagtgat cagccagctg cacgctgcca tcctccacgt
     3241 tgtggcgaat tttaaaattc gctttaatgc catttttttg tttatcggcg gt
//
