LOCUS       sequence-439532-        7288 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the S. cerevisiae SUC2 (Invertase) protein (Part3b') with 
            the Aga2 signal peptide (Part3a') assembled with pTEF1 promoter, 
            tTDH1 terminator, LEU2 selectable marker and CEN/ARS yeast origin..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7288)
  AUTHORS   O'Riordan NM, Juric V, O'Neill SK, Roche AP, Young PW
  TITLE     A Yeast Modular Cloning (MoClo) Toolkit Expansion for Optimization 
            of Heterologous Protein Secretion and Surface Display in 
            Saccharomyces cerevisiae.
  JOURNAL   ACS Synth Biol. 2024 Mar 14. doi: 10.1021/acssynbio.3c00743.
  PUBMED    38483353
REFERENCE   2  (bases 1 to 7288)
  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. 2024 Mar 14. doi: 10.1021/acssynbio.3c00743."
COMMENT     Sequence Label: pYSD101
FEATURES             Location/Qualifiers
     source          1..7288
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    531..1034
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(join(1145..1936,1937..2005))
                     /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     1768..1787
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2006..2110)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2213..2801
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2702..2721
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3380..3793
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             5391..5462
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             5466..5483
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        6138..6545
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             join(6546..7288,1..352)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
ORIGIN
        1 tggtgatatc atctccgatg aagcctccgt tatcccaggt tccttgggtt tgttgccatc
       61 tgcgtccttg gcctctttgc cagacaagaa caccgcattt ggtttgtacg aaccatgcca
      121 cggttctgct ccagatttgc caaagaataa ggtcaaccct atcgccacta tcttgtctgc
      181 tgcaatgatg ttgaaattgt cattgaactt gcctgaagaa ggtaaggcca ttgaagatgc
      241 agttaaaaag gttttggatg caggtatcag aactggtgat ttaggtggtt ccaacagtac
      301 caccgaagtc ggtgatgctg tcgccgaaga agttaagaaa atccttgctt aacctggagg
      361 acccttctct ttagactatt ctactcttat gcacgtaaaa aattctagga aatatgtatt
      421 aactaggagt aaaataaccg gctagtggca ttcatatagc cgtctgttta catctacatc
      481 acacatttcg agtgtatatc tcgcaacgtt ggcgttaaat aggcaggagt atcacgtgct
      541 ataaaaataa ttataattta aattttttaa tataaatata taaattaaaa atagaaagta
      601 aaaaaagaaa ttaaagaaaa aatagttttt gttttccgaa gatgtaaaag actctagggg
      661 gatcgccaac aaatactacc ttttatcttg ctcttcctgc tctcaggtat taatgccgaa
      721 ttgtttcatc ttgtctgtgt agaaaaccac acacgaaaat cctgtgattt tacattttac
      781 ttatcgttaa tcgaatgtat atctatttaa tctgcttttc ttgtctaata aatatatatg
      841 taaagtacgc tttttgttga aattttttaa acctttgttt attttttttt cttcattccg
      901 taactcttct accttcttta tttactttct aaaatccaaa tacaaaacat aaaaataaat
      961 aaacacagag taaattccca aattattcca tcattaaaag atacgaggcg cgtgtaagtt
     1021 acaggcaagc gatccgtccc gagcggccgc gattatcaaa aaggatcttc acctagatcc
     1081 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1141 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1201 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1261 gccccagtgc tgcaatgata ccgcgggacc cacgctcacc ggctccagat ttatcagcaa
     1321 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1381 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1441 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1501 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1561 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1621 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1681 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     1741 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1801 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     1861 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     1921 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     1981 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     2041 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2101 gggttccgcg cacatttccc cgaaaagtgc cacctgtcat gaccaaaatc ccttaacgtg
     2161 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2221 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2281 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2341 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2401 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2461 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     2521 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     2581 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     2641 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     2701 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     2761 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2821 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     2881 ctgattctgt ggataaccgt gcggccgccc ctgaattcgc atctagaacg gggtcatcac
     2941 ggctcatcat gcgccaaaca aatgtgtgca atacacgctc ggatgactgc atgatgaccg
     3001 cactgactgg ggacagcaga tccacctaag cctgtgagag aagcagacac ccgacagatc
     3061 aaggcagtta cgtctcacca aaccagatgt caacacagct acaacgcctt gccaacaggg
     3121 agttcttcag agacatggag gctcaaaacg aaattattga cagcctagac atcaatagtc
     3181 atacaacaga aagcgaccac ccaactttgg ctgataatag cgtataaaca atgcatactt
     3241 tgtacgttca aaatacaatg cagtagatat atttatgcat attacatata atacatatca
     3301 cataggaagc aacaggcgcg ttggactttt aattttcgag gaccgcgaat ccttacatca
     3361 cacccaatcc cccacaagtg atcccccaca caccatagct tcaaaatgtt tctactcctt
     3421 ttttactctt ccagattttc tcggactccg cgcatcgccg taccacttca aaacacccaa
     3481 gcacagcata ctaaatttcc cctctttctt cctctagggt gtcgttaatt acccgtacta
     3541 aaggtttgga aaagaaaaaa gacaccgcct cgtttctttt tcttcgtcga aaaaggcaat
     3601 aaaaattttt atcacgtttc tttttcttga aaattttttt ttttgatttt tttctctttc
     3661 gatgacctcc cattgatatt taagttaata aacggtcatc aatttctcaa gtttcagttt
     3721 catttttctt gttctattac aacttttttt acttcttgct cattagaaag aaagcatagc
     3781 aatctaatct aagttttaat tacaaaagat ctatgatgca gttacttcgc tgtttttcaa
     3841 tattttctgt tattgcttca gttcttgctt caatgacaaa cgaaactagc gatagacctt
     3901 tggtccactt cacacccaac aagggctgga tgaatgaccc aaatgggttg tggtacgatg
     3961 aaaaagatgc caaatggcat ctgtactttc aatacaaccc aaatgacacc gtatggggta
     4021 cgccattgtt ttggggccat gctacttccg atgatttgac taattgggaa gatcaaccca
     4081 ttgctatcgc tcccaagcgt aacgattcag gtgctttctc tggctccatg gtggttgatt
     4141 acaacaacac gagtgggttt ttcaatgata ctattgatcc aagacaaaga tgcgttgcga
     4201 tttggactta taacactcct gaaagtgaag agcaatacat tagctattct cttgatggtg
     4261 gttacacttt tactgaatac caaaagaacc ctgttttagc tgccaactcc actcaattca
     4321 gagatccaaa ggtgttctgg tatgaacctt ctcaaaaatg gattatgacg gctgccaaat
     4381 cacaagacta caaaattgaa atttactcct ctgatgactt gaagtcctgg aagctagaat
     4441 ctgcatttgc caatgaaggt ttcttaggct accaatacga atgtccaggt ttgattgaag
     4501 tcccaactga gcaagatcct tccaaatctt attgggtcat gtttatttct atcaacccag
     4561 gtgcacctgc tggcggttcc ttcaaccaat attttgttgg atccttcaat ggtactcatt
     4621 ttgaagcgtt tgacaatcaa tctagagtgg tagattttgg taaggactac tatgccttgc
     4681 aaactttctt caacactgac ccaacctacg gttcagcatt aggtattgcc tgggcttcaa
     4741 actgggagta cagtgccttt gtcccaacta acccatggag atcatccatg tctttggtcc
     4801 gcaagttttc tttgaacact gaatatcaag ctaatccaga gactgaattg atcaatttga
     4861 aagccgaacc aatattgaac attagtaatg ctggtccctg gtcgcgtttt gctactaaca
     4921 caactctaac taaggccaat tcttacaatg tcgatttgag caactcgact ggtaccctag
     4981 agtttgagtt ggtttacgct gttaacacca cacaaaccat atccaaatcc gtctttgccg
     5041 acttatcact ttggttcaag ggtttagaag atcctgaaga atatttgaga atgggttttg
     5101 aagtcagtgc ttcttccttc tttttggacc gtggtaactc taaggtcaag tttgtcaagg
     5161 agaacccata tttcacaaac agaatgtctg tcaacaacca accattcaag tctgagaacg
     5221 acctaagtta ctataaagtg tacggcctac tggatcaaaa catcttggaa ttgtacttca
     5281 acgatggaga tgtggtttct acaaatacct acttcatgac caccggtaac gctctaggat
     5341 ctgtgaacat gaccactggt gtcgataatt tgttctacat tgacggatcc gattacaagg
     5401 acgatgatga taaggactac aaagatgatg atgacaaaga ttataaggac gatgatgata
     5461 aaggtcatca tcatcatcat cactaactcg agtggcataa agcaatcttg atgaggataa
     5521 tgattttttt ttgaatatac ataaatacta ccgtttttct gctagatttt gtgatgacgt
     5581 aaataagtac atattacttt ttaagccaag acaagattaa gcattaactt tacccttttc
     5641 tttctaagtt tcaatattag ttatcactgt ttaaaagtta tggcgagaac gtcggcggtt
     5701 aaaatatatt accctgaacg gctggatgtg agacgataaa tcgcagccaa gtgagtgaat
     5761 agatgacgca ccacggtcag acacggccac atcgtatctc acaggagcaa gcgcgatagg
     5821 agcactcaca catagtacgg tgatccgctg actcctttgc ccaaataaga cgtgagccac
     5881 tagtgcactg cagtacatcg aggagaactt ctagtatatc tacataccta atattattgc
     5941 cttattaaaa atggaatccc aacaattaca tcaaaatcca cattctcttc aaaatcaatt
     6001 gtcctgtact tccttgttca tgtgtgttca aaaacgttat atttatagga taattatact
     6061 ctatttctca acaagtaatt ggttgtttgg ccgagcggtc taaggcgcct gattcaagaa
     6121 atatcttgac cgcagttaac tgtgggaata ctcaggtatc gtaagatgca agagttcgaa
     6181 tctcttagca accattattt ttttcctcaa cataacgaga acacacaggg gcgctatcgc
     6241 acagaatcaa attcgatgac tggaaatttt ttgttaattt cagaggtcgc ctgacgcata
     6301 tacctttttc aactgaaaaa ttgggagaaa aaggaaaggt gagagcgccg gaaccggctt
     6361 ttcatataga atagagaagc gttcatgact aaatgcttgc atcacaatac ttgaagttga
     6421 caatattatt taaggaccta ttgttttttc caataggtgg ttagcaatcg tcttactttc
     6481 taacttttct taccttttac atttcagcaa tatatatata tatatttcaa ggatatacca
     6541 ttctaatgtc tgcccctaag aagatcgtcg ttttgccagg tgaccacgtt ggtcaagaaa
     6601 tcacagccga agccattaag gttcttaaag ctatttctga tgttcgttcc aatgtcaagt
     6661 tcgatttcga aaatcattta attggtggtg ctgctatcga tgctacaggt gttccacttc
     6721 cagatgaggc gctggaagcc tccaagaagg ctgatgccgt tttgttaggt gctgtgggtg
     6781 gtcctaaatg gggaaccggt agtgttagac ctgaacaagg tttactaaaa atccgtaaag
     6841 aacttcaatt gtacgccaac ttaagaccat gtaactttgc atccgactct cttttagact
     6901 tatctccaat caagccacaa tttgctaaag gtactgactt cgttgttgtc agagaattag
     6961 tgggaggtat ttactttggt aagagaaagg aggacgatgg tgatggtgtc gcttgggata
     7021 gtgaacaata caccgttcca gaagtgcaaa gaatcacaag aatggccgct ttcatggccc
     7081 tacaacatga gccaccattg cctatttggt ccttggataa agctaatgtt ttggcctctt
     7141 caagattatg gagaaaaact gtggaggaaa ccatcaagaa cgaatttcct acattgaagg
     7201 ttcaacatca attgattgat tctgccgcca tgatcctagt taagaaccca acccacctaa
     7261 atggtattat aatcaccagc aacatgtt
//
