LOCUS       sequence-421716-        7234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the S. cerevisiae SUC2 (Invertase) protein (Part3) without 
            any signal peptide 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 7234)
  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 7234)
  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: pYSD100
FEATURES             Location/Qualifiers
     source          1..7234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        472..885
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             2429..2500
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             2504..2521
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        3176..3583
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             3584..4678
                     /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"
     misc_feature    4857..5360
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(join(5471..6262,6263..6331))
                     /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     6094..6113
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6332..6436)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6539..7127
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7028..7047
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgaattc gcatctagaa cggggtcatc acggctcatc atgcgccaaa caaatgtgtg
       61 caatacacgc tcggatgact gcatgatgac cgcactgact ggggacagca gatccaccta
      121 agcctgtgag agaagcagac acccgacaga tcaaggcagt tacgtctcac caaaccagat
      181 gtcaacacag ctacaacgcc ttgccaacag ggagttcttc agagacatgg aggctcaaaa
      241 cgaaattatt gacagcctag acatcaatag tcatacaaca gaaagcgacc acccaacttt
      301 ggctgataat agcgtataaa caatgcatac tttgtacgtt caaaatacaa tgcagtagat
      361 atatttatgc atattacata taatacatat cacataggaa gcaacaggcg cgttggactt
      421 ttaattttcg aggaccgcga atccttacat cacacccaat cccccacaag tgatccccca
      481 cacaccatag cttcaaaatg tttctactcc ttttttactc ttccagattt tctcggactc
      541 cgcgcatcgc cgtaccactt caaaacaccc aagcacagca tactaaattt cccctctttc
      601 ttcctctagg gtgtcgttaa ttacccgtac taaaggtttg gaaaagaaaa aagacaccgc
      661 ctcgtttctt tttcttcgtc gaaaaaggca ataaaaattt ttatcacgtt tctttttctt
      721 gaaaattttt ttttttgatt tttttctctt tcgatgacct cccattgata tttaagttaa
      781 taaacggtca tcaatttctc aagtttcagt ttcatttttc ttgttctatt acaacttttt
      841 ttacttcttg ctcattagaa agaaagcata gcaatctaat ctaagtttta attacaaaag
      901 atctatgtca atgacaaacg aaactagcga tagacctttg gtccacttca cacccaacaa
      961 gggctggatg aatgacccaa atgggttgtg gtacgatgaa aaagatgcca aatggcatct
     1021 gtactttcaa tacaacccaa atgacaccgt atggggtacg ccattgtttt ggggccatgc
     1081 tacttccgat gatttgacta attgggaaga tcaacccatt gctatcgctc ccaagcgtaa
     1141 cgattcaggt gctttctctg gctccatggt ggttgattac aacaacacga gtgggttttt
     1201 caatgatact attgatccaa gacaaagatg cgttgcgatt tggacttata acactcctga
     1261 aagtgaagag caatacatta gctattctct tgatggtggt tacactttta ctgaatacca
     1321 aaagaaccct gttttagctg ccaactccac tcaattcaga gatccaaagg tgttctggta
     1381 tgaaccttct caaaaatgga ttatgacggc tgccaaatca caagactaca aaattgaaat
     1441 ttactcctct gatgacttga agtcctggaa gctagaatct gcatttgcca atgaaggttt
     1501 cttaggctac caatacgaat gtccaggttt gattgaagtc ccaactgagc aagatccttc
     1561 caaatcttat tgggtcatgt ttatttctat caacccaggt gcacctgctg gcggttcctt
     1621 caaccaatat tttgttggat ccttcaatgg tactcatttt gaagcgtttg acaatcaatc
     1681 tagagtggta gattttggta aggactacta tgccttgcaa actttcttca acactgaccc
     1741 aacctacggt tcagcattag gtattgcctg ggcttcaaac tgggagtaca gtgcctttgt
     1801 cccaactaac ccatggagat catccatgtc tttggtccgc aagttttctt tgaacactga
     1861 atatcaagct aatccagaga ctgaattgat caatttgaaa gccgaaccaa tattgaacat
     1921 tagtaatgct ggtccctggt cgcgttttgc tactaacaca actctaacta aggccaattc
     1981 ttacaatgtc gatttgagca actcgactgg taccctagag tttgagttgg tttacgctgt
     2041 taacaccaca caaaccatat ccaaatccgt ctttgccgac ttatcacttt ggttcaaggg
     2101 tttagaagat cctgaagaat atttgagaat gggttttgaa gtcagtgctt cttccttctt
     2161 tttggaccgt ggtaactcta aggtcaagtt tgtcaaggag aacccatatt tcacaaacag
     2221 aatgtctgtc aacaaccaac cattcaagtc tgagaacgac ctaagttact ataaagtgta
     2281 cggcctactg gatcaaaaca tcttggaatt gtacttcaac gatggagatg tggtttctac
     2341 aaatacctac ttcatgacca ccggtaacgc tctaggatct gtgaacatga ccactggtgt
     2401 cgataatttg ttctacattg acggatccga ttacaaggac gatgatgata aggactacaa
     2461 agatgatgat gacaaagatt ataaggacga tgatgataaa ggtcatcatc atcatcatca
     2521 ctaactcgag tggcataaag caatcttgat gaggataatg attttttttt gaatatacat
     2581 aaatactacc gtttttctgc tagattttgt gatgacgtaa ataagtacat attacttttt
     2641 aagccaagac aagattaagc attaacttta cccttttctt tctaagtttc aatattagtt
     2701 atcactgttt aaaagttatg gcgagaacgt cggcggttaa aatatattac cctgaacggc
     2761 tggatgtgag acgataaatc gcagccaagt gagtgaatag atgacgcacc acggtcagac
     2821 acggccacat cgtatctcac aggagcaagc gcgataggag cactcacaca tagtacggtg
     2881 atccgctgac tcctttgccc aaataagacg tgagccacta gtgcactgca gtacatcgag
     2941 gagaacttct agtatatcta catacctaat attattgcct tattaaaaat ggaatcccaa
     3001 caattacatc aaaatccaca ttctcttcaa aatcaattgt cctgtacttc cttgttcatg
     3061 tgtgttcaaa aacgttatat ttataggata attatactct atttctcaac aagtaattgg
     3121 ttgtttggcc gagcggtcta aggcgcctga ttcaagaaat atcttgaccg cagttaactg
     3181 tgggaatact caggtatcgt aagatgcaag agttcgaatc tcttagcaac cattattttt
     3241 ttcctcaaca taacgagaac acacaggggc gctatcgcac agaatcaaat tcgatgactg
     3301 gaaatttttt gttaatttca gaggtcgcct gacgcatata cctttttcaa ctgaaaaatt
     3361 gggagaaaaa ggaaaggtga gagcgccgga accggctttt catatagaat agagaagcgt
     3421 tcatgactaa atgcttgcat cacaatactt gaagttgaca atattattta aggacctatt
     3481 gttttttcca ataggtggtt agcaatcgtc ttactttcta acttttctta ccttttacat
     3541 ttcagcaata tatatatata tatttcaagg atataccatt ctaatgtctg cccctaagaa
     3601 gatcgtcgtt ttgccaggtg accacgttgg tcaagaaatc acagccgaag ccattaaggt
     3661 tcttaaagct atttctgatg ttcgttccaa tgtcaagttc gatttcgaaa atcatttaat
     3721 tggtggtgct gctatcgatg ctacaggtgt tccacttcca gatgaggcgc tggaagcctc
     3781 caagaaggct gatgccgttt tgttaggtgc tgtgggtggt cctaaatggg gaaccggtag
     3841 tgttagacct gaacaaggtt tactaaaaat ccgtaaagaa cttcaattgt acgccaactt
     3901 aagaccatgt aactttgcat ccgactctct tttagactta tctccaatca agccacaatt
     3961 tgctaaaggt actgacttcg ttgttgtcag agaattagtg ggaggtattt actttggtaa
     4021 gagaaaggag gacgatggtg atggtgtcgc ttgggatagt gaacaataca ccgttccaga
     4081 agtgcaaaga atcacaagaa tggccgcttt catggcccta caacatgagc caccattgcc
     4141 tatttggtcc ttggataaag ctaatgtttt ggcctcttca agattatgga gaaaaactgt
     4201 ggaggaaacc atcaagaacg aatttcctac attgaaggtt caacatcaat tgattgattc
     4261 tgccgccatg atcctagtta agaacccaac ccacctaaat ggtattataa tcaccagcaa
     4321 catgtttggt gatatcatct ccgatgaagc ctccgttatc ccaggttcct tgggtttgtt
     4381 gccatctgcg tccttggcct ctttgccaga caagaacacc gcatttggtt tgtacgaacc
     4441 atgccacggt tctgctccag atttgccaaa gaataaggtc aaccctatcg ccactatctt
     4501 gtctgctgca atgatgttga aattgtcatt gaacttgcct gaagaaggta aggccattga
     4561 agatgcagtt aaaaaggttt tggatgcagg tatcagaact ggtgatttag gtggttccaa
     4621 cagtaccacc gaagtcggtg atgctgtcgc cgaagaagtt aagaaaatcc ttgcttaacc
     4681 tggaggaccc ttctctttag actattctac tcttatgcac gtaaaaaatt ctaggaaata
     4741 tgtattaact aggagtaaaa taaccggcta gtggcattca tatagccgtc tgtttacatc
     4801 tacatcacac atttcgagtg tatatctcgc aacgttggcg ttaaataggc aggagtatca
     4861 cgtgctataa aaataattat aatttaaatt ttttaatata aatatataaa ttaaaaatag
     4921 aaagtaaaaa aagaaattaa agaaaaaata gtttttgttt tccgaagatg taaaagactc
     4981 tagggggatc gccaacaaat actacctttt atcttgctct tcctgctctc aggtattaat
     5041 gccgaattgt ttcatcttgt ctgtgtagaa aaccacacac gaaaatcctg tgattttaca
     5101 ttttacttat cgttaatcga atgtatatct atttaatctg cttttcttgt ctaataaata
     5161 tatatgtaaa gtacgctttt tgttgaaatt ttttaaacct ttgtttattt ttttttcttc
     5221 attccgtaac tcttctacct tctttattta ctttctaaaa tccaaataca aaacataaaa
     5281 ataaataaac acagagtaaa ttcccaaatt attccatcat taaaagatac gaggcgcgtg
     5341 taagttacag gcaagcgatc cgtcccgagc ggccgcgatt atcaaaaagg atcttcacct
     5401 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5461 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5521 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5581 catctggccc cagtgctgca atgataccgc gggacccacg ctcaccggct ccagatttat
     5641 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5701 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5761 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5821 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5881 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5941 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6001 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6061 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6121 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6181 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6241 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6301 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6361 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6421 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgtcatgacc aaaatccctt
     6481 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6541 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6601 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6661 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6721 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6781 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6841 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6901 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6961 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     7021 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     7081 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     7141 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     7201 tatcccctga ttctgtggat aaccgtgcgg ccgc
//
