LOCUS       sequence-424921-        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"
     CDS             570..641
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             645..662
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        1317..1724
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             1725..2819
                     /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    2998..3501
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(join(3612..4403,4404..4472))
                     /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     4235..4254
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4473..4577)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4680..5268
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5169..5188
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5847..6260
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 aatatcaagc taatccagag actgaattga tcaatttgaa agccgaacca atattgaaca
       61 ttagtaatgc tggtccctgg tcgcgttttg ctactaacac aactctaact aaggccaatt
      121 cttacaatgt cgatttgagc aactcgactg gtaccctaga gtttgagttg gtttacgctg
      181 ttaacaccac acaaaccata tccaaatccg tctttgccga cttatcactt tggttcaagg
      241 gtttagaaga tcctgaagaa tatttgagaa tgggttttga agtcagtgct tcttccttct
      301 ttttggaccg tggtaactct aaggtcaagt ttgtcaagga gaacccatat ttcacaaaca
      361 gaatgtctgt caacaaccaa ccattcaagt ctgagaacga cctaagttac tataaagtgt
      421 acggcctact ggatcaaaac atcttggaat tgtacttcaa cgatggagat gtggtttcta
      481 caaataccta cttcatgacc accggtaacg ctctaggatc tgtgaacatg accactggtg
      541 tcgataattt gttctacatt gacggatccg attacaagga cgatgatgat aaggactaca
      601 aagatgatga tgacaaagat tataaggacg atgatgataa aggtcatcat catcatcatc
      661 actaactcga gtggcataaa gcaatcttga tgaggataat gatttttttt tgaatataca
      721 taaatactac cgtttttctg ctagattttg tgatgacgta aataagtaca tattactttt
      781 taagccaaga caagattaag cattaacttt acccttttct ttctaagttt caatattagt
      841 tatcactgtt taaaagttat ggcgagaacg tcggcggtta aaatatatta ccctgaacgg
      901 ctggatgtga gacgataaat cgcagccaag tgagtgaata gatgacgcac cacggtcaga
      961 cacggccaca tcgtatctca caggagcaag cgcgatagga gcactcacac atagtacggt
     1021 gatccgctga ctcctttgcc caaataagac gtgagccact agtgcactgc agtacatcga
     1081 ggagaacttc tagtatatct acatacctaa tattattgcc ttattaaaaa tggaatccca
     1141 acaattacat caaaatccac attctcttca aaatcaattg tcctgtactt ccttgttcat
     1201 gtgtgttcaa aaacgttata tttataggat aattatactc tatttctcaa caagtaattg
     1261 gttgtttggc cgagcggtct aaggcgcctg attcaagaaa tatcttgacc gcagttaact
     1321 gtgggaatac tcaggtatcg taagatgcaa gagttcgaat ctcttagcaa ccattatttt
     1381 tttcctcaac ataacgagaa cacacagggg cgctatcgca cagaatcaaa ttcgatgact
     1441 ggaaattttt tgttaatttc agaggtcgcc tgacgcatat acctttttca actgaaaaat
     1501 tgggagaaaa aggaaaggtg agagcgccgg aaccggcttt tcatatagaa tagagaagcg
     1561 ttcatgacta aatgcttgca tcacaatact tgaagttgac aatattattt aaggacctat
     1621 tgttttttcc aataggtggt tagcaatcgt cttactttct aacttttctt accttttaca
     1681 tttcagcaat atatatatat atatttcaag gatataccat tctaatgtct gcccctaaga
     1741 agatcgtcgt tttgccaggt gaccacgttg gtcaagaaat cacagccgaa gccattaagg
     1801 ttcttaaagc tatttctgat gttcgttcca atgtcaagtt cgatttcgaa aatcatttaa
     1861 ttggtggtgc tgctatcgat gctacaggtg ttccacttcc agatgaggcg ctggaagcct
     1921 ccaagaaggc tgatgccgtt ttgttaggtg ctgtgggtgg tcctaaatgg ggaaccggta
     1981 gtgttagacc tgaacaaggt ttactaaaaa tccgtaaaga acttcaattg tacgccaact
     2041 taagaccatg taactttgca tccgactctc ttttagactt atctccaatc aagccacaat
     2101 ttgctaaagg tactgacttc gttgttgtca gagaattagt gggaggtatt tactttggta
     2161 agagaaagga ggacgatggt gatggtgtcg cttgggatag tgaacaatac accgttccag
     2221 aagtgcaaag aatcacaaga atggccgctt tcatggccct acaacatgag ccaccattgc
     2281 ctatttggtc cttggataaa gctaatgttt tggcctcttc aagattatgg agaaaaactg
     2341 tggaggaaac catcaagaac gaatttccta cattgaaggt tcaacatcaa ttgattgatt
     2401 ctgccgccat gatcctagtt aagaacccaa cccacctaaa tggtattata atcaccagca
     2461 acatgtttgg tgatatcatc tccgatgaag cctccgttat cccaggttcc ttgggtttgt
     2521 tgccatctgc gtccttggcc tctttgccag acaagaacac cgcatttggt ttgtacgaac
     2581 catgccacgg ttctgctcca gatttgccaa agaataaggt caaccctatc gccactatct
     2641 tgtctgctgc aatgatgttg aaattgtcat tgaacttgcc tgaagaaggt aaggccattg
     2701 aagatgcagt taaaaaggtt ttggatgcag gtatcagaac tggtgattta ggtggttcca
     2761 acagtaccac cgaagtcggt gatgctgtcg ccgaagaagt taagaaaatc cttgcttaac
     2821 ctggaggacc cttctcttta gactattcta ctcttatgca cgtaaaaaat tctaggaaat
     2881 atgtattaac taggagtaaa ataaccggct agtggcattc atatagccgt ctgtttacat
     2941 ctacatcaca catttcgagt gtatatctcg caacgttggc gttaaatagg caggagtatc
     3001 acgtgctata aaaataatta taatttaaat tttttaatat aaatatataa attaaaaata
     3061 gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt ttccgaagat gtaaaagact
     3121 ctagggggat cgccaacaaa tactaccttt tatcttgctc ttcctgctct caggtattaa
     3181 tgccgaattg tttcatcttg tctgtgtaga aaaccacaca cgaaaatcct gtgattttac
     3241 attttactta tcgttaatcg aatgtatatc tatttaatct gcttttcttg tctaataaat
     3301 atatatgtaa agtacgcttt ttgttgaaat tttttaaacc tttgtttatt tttttttctt
     3361 cattccgtaa ctcttctacc ttctttattt actttctaaa atccaaatac aaaacataaa
     3421 aataaataaa cacagagtaa attcccaaat tattccatca ttaaaagata cgaggcgcgt
     3481 gtaagttaca ggcaagcgat ccgtcccgag cggccgcgat tatcaaaaag gatcttcacc
     3541 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3601 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3661 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3721 ccatctggcc ccagtgctgc aatgataccg cgggacccac gctcaccggc tccagattta
     3781 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3841 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3901 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3961 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4021 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4081 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4141 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4201 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4261 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4321 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4381 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4441 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4501 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4561 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgtcatgac caaaatccct
     4621 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4681 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4741 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4801 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4861 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4921 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4981 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5041 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5101 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5161 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5221 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5281 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5341 ttatcccctg attctgtgga taaccgtgcg gccgcccctg aattcgcatc tagaacgggg
     5401 tcatcacggc tcatcatgcg ccaaacaaat gtgtgcaata cacgctcgga tgactgcatg
     5461 atgaccgcac tgactgggga cagcagatcc acctaagcct gtgagagaag cagacacccg
     5521 acagatcaag gcagttacgt ctcaccaaac cagatgtcaa cacagctaca acgccttgcc
     5581 aacagggagt tcttcagaga catggaggct caaaacgaaa ttattgacag cctagacatc
     5641 aatagtcata caacagaaag cgaccaccca actttggctg ataatagcgt ataaacaatg
     5701 catactttgt acgttcaaaa tacaatgcag tagatatatt tatgcatatt acatataata
     5761 catatcacat aggaagcaac aggcgcgttg gacttttaat tttcgaggac cgcgaatcct
     5821 tacatcacac ccaatccccc acaagtgatc ccccacacac catagcttca aaatgtttct
     5881 actccttttt tactcttcca gattttctcg gactccgcgc atcgccgtac cacttcaaaa
     5941 cacccaagca cagcatacta aatttcccct ctttcttcct ctagggtgtc gttaattacc
     6001 cgtactaaag gtttggaaaa gaaaaaagac accgcctcgt ttctttttct tcgtcgaaaa
     6061 aggcaataaa aatttttatc acgtttcttt ttcttgaaaa tttttttttt tgattttttt
     6121 ctctttcgat gacctcccat tgatatttaa gttaataaac ggtcatcaat ttctcaagtt
     6181 tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgctcat tagaaagaaa
     6241 gcatagcaat ctaatctaag ttttaattac aaaagatcta tgatgcagtt acttcgctgt
     6301 ttttcaatat tttctgttat tgcttcagtt cttgcttcaa tgacaaacga aactagcgat
     6361 agacctttgg tccacttcac acccaacaag ggctggatga atgacccaaa tgggttgtgg
     6421 tacgatgaaa aagatgccaa atggcatctg tactttcaat acaacccaaa tgacaccgta
     6481 tggggtacgc cattgttttg gggccatgct acttccgatg atttgactaa ttgggaagat
     6541 caacccattg ctatcgctcc caagcgtaac gattcaggtg ctttctctgg ctccatggtg
     6601 gttgattaca acaacacgag tgggtttttc aatgatacta ttgatccaag acaaagatgc
     6661 gttgcgattt ggacttataa cactcctgaa agtgaagagc aatacattag ctattctctt
     6721 gatggtggtt acacttttac tgaataccaa aagaaccctg ttttagctgc caactccact
     6781 caattcagag atccaaaggt gttctggtat gaaccttctc aaaaatggat tatgacggct
     6841 gccaaatcac aagactacaa aattgaaatt tactcctctg atgacttgaa gtcctggaag
     6901 ctagaatctg catttgccaa tgaaggtttc ttaggctacc aatacgaatg tccaggtttg
     6961 attgaagtcc caactgagca agatccttcc aaatcttatt gggtcatgtt tatttctatc
     7021 aacccaggtg cacctgctgg cggttccttc aaccaatatt ttgttggatc cttcaatggt
     7081 actcattttg aagcgtttga caatcaatct agagtggtag attttggtaa ggactactat
     7141 gccttgcaaa ctttcttcaa cactgaccca acctacggtt cagcattagg tattgcctgg
     7201 gcttcaaact gggagtacag tgcctttgtc ccaactaacc catggagatc atccatgtct
     7261 ttggtccgca agttttcttt gaacactg
//
