LOCUS       sequence-421718-        6992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the S. cerevisiae Aga1 protein (Part3) with pGBK1 promoter, 
            tENO2 terminator, HIS3 selectable marker and CEN/ARS yeast origin..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6992)
  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 6992)
  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: pYSD300
FEATURES             Location/Qualifiers
     source          1..6992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3550..3737
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             3738..4400
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     misc_feature    4615..5118
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(join(5229..6020,6021..6089))
                     /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     5852..5871
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6090..6194)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6297..6885
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6786..6805
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgaattc gcatctagat ggtagagcca caaacagccg gtacaagcaa cgatctccag
       61 gaccatctga atcatgcgcg gatgacacga actcacgacg gcgatcacag acattaaccc
      121 acagtacaga cactgcgaca acgtggcaat tcgtcgcaat accgtctcac tgaactggcc
      181 gataattgca gacgaacggt gagtaaggaa agagtgagga actatcgcat acctgcattt
      241 aaagatgccg atttgggcgc gaatccttta ttttggcttc accctcatac tattatcagg
      301 gccagaaaaa ggaagtgttt ccctccttct tgaattgatg ttaccctcat aaagcacgtg
      361 gcctcttatc gagaaagaaa ttaccgtcgc tcgtgatttg tttgcaaaaa gaacaaaact
      421 gaaaaaaccc agacacgctc gacttcctgt catcctattg attgcagctt ccaatttcgt
      481 cacacaacaa ggtcctagcg acggctcaca ggttttgtaa caagcaatcg aaggttctgg
      541 aatggcggga aagggtttag taccacatgc tatgatgccc actgtgatct ccagagcaaa
      601 gttcgttcga tcgtactgtt actctctctc tttcaaacag aattgtccga atcgtgtgac
      661 aacaacagcc tgttctcaca cactcttttc ttctaaccaa gggggtggtt tagtttagta
      721 gaacctcgtg aaacttacat ttacatatat ataaacttgc ataaattggt caatgcaaga
      781 aatacatatt tggtcttttc taattcgtag tttttcaagt tcttagatgc tttctttttc
      841 tcttttttac agatcatcaa ggaagtaatt atctactttt tacaacaaat ataaaacaag
      901 atctatgaca ttatctttcg ctcattttac ctacctgttc acaatattgt tgggattaac
      961 taatattgcc ttggcatctg atccagaaac gattctagtg acgataacca agacaaacga
     1021 tgcaaatggg gttgttacaa ctacagtttc acccgcgcta gtctccacat ccactatcgt
     1081 tcaagctggc actacgacat tgtatacgac ttggtgtcca ttgacggtat ccacttcatc
     1141 tgctgccgaa ataagtcctt caatatcgta cgctactacc ctatccagat ttagtacttt
     1201 gacattatct acagaagtct gctcccatga ggcatgtcct tcgtcatcga cgttgccaac
     1261 caccacctta tctgtgactt ccaagttcac ttcatatatt tgccctactt gtcacacaac
     1321 cgctatcagc tcattatccg aagtaggaac tacaaccgtg gtatcatcca gcgccattga
     1381 accatcaagt gcctctataa tctcacctgt cacctctaca ctttcgagta caacatcgtc
     1441 caatccaact actacctccc taagttcgac atctacatct ccaagctcta catctacatc
     1501 tccaagctct acatctacct catcaagttc gacatctacc tcatcaagtt cgacatctac
     1561 ctcatcaagt tcgacatcta catctccaag ttcgacatcc acatcttcaa gtttgacatc
     1621 cacatcttca agttctacat ctacatccca aagttctaca tctacctcat caagttcgac
     1681 atctacatct ccaagctcta catctacctc atcaagttca acatctacat ctccaagttc
     1741 taaatctact tctgcaagct ccacttccac ttcttcatat tcaacatcta catccccaag
     1801 tttgacttct tcatctccaa ctttggcttc cacttctcca agttcaacat ctattagctc
     1861 tacttttact gattcaactt catcccttgg ctcctctata gcatcttcat caacgtctgt
     1921 gtcattatac agcccatcca cacctgttta ctccgtccct tcgacttcgt caaatgttgc
     1981 aactccttct atgacttctt caactgttga aacaactgtt agttcacaaa gttcgtctga
     2041 atatatcacc aaatcctcaa tttctactac tatcccatca ttttccatgt ctacatattt
     2101 caccactgtt agtggagtca ctacaatgta tacgacatgg tgtccttata gctctgaatc
     2161 tgagactagc acattaacca gtatgcatga aacggttaca acagacgcta cagtctgcac
     2221 tcacgagtct tgcatgccct cgcagacaac aagtttgatt acatcttcta taaaaatgtc
     2281 cactaaaaac gtcgcaactt ctgtaagcac ctcaacggtt gaatcctcat atgcatgctc
     2341 cacatgtgct gaaacgtcac actcgtattc ttccgtgcaa acagcttcat caagttctgt
     2401 aacacagcag accacatcca caaagagttg ggtaagttca atgacaactt cggatgaaga
     2461 tttcaataag cacgctaccg gtaagtatca tgtaacatct tcaggtacct caaccatttc
     2521 gactagtgta agtgaagcca cgagtacatc aagcattgac tcagaatctc aagaacaatc
     2581 atcacactta ttatcgacat cggtcctttc atcctcctcc ttgtctgcta cattatcctc
     2641 tgacagtact attttgctat tcagttctgt atcatcacta agtgtcgaac agtcaccagt
     2701 taccacactt caaatttctt caacatcaga gattttacaa cccacttctt ccacagctat
     2761 tgctacaata tctgcctcta catcatcact ttccgcaaca tctatctcta caccatctac
     2821 ctctgtggaa tcgactattg aatcttcatc attgactccg acggtatctt ctattttcct
     2881 ctcatcatca tctgctccct cttctctaca aacatctgtt accactacag aagtttccac
     2941 tacttcaatc tccatacaat accaaacttc atcaatggta acaattagcc aatatatggg
     3001 cagtggatcg caaacgcgtt tgccattagg aaagttggtc ttcgccatca tggcagttgc
     3061 ttgcaatgta attttcagtt aataaatcct aactcgagag tgcttttaac taagaattat
     3121 tagtcttttc tgcttatttt ttcatcatag tttagaacac tttatattaa cgaatagttt
     3181 atgaatctat ttaggtttaa aaattgatac agttttataa gttacttttt caaagactcg
     3241 tgctgtctat tgcataatgc actggaaggg gaaaaaaaag gtgcacacgc gtggcttttt
     3301 cttgaatttg cagtttgaaa aatgctgcca atgagacgac ggggtcatca cggctcatca
     3361 tgcgccaaac aaatgtgtgc aatacacgct cggatgactg catgatgacc gcactgactg
     3421 gggacagcag atccacctaa gcctgtgaga gaagcagaca cccgacagat caaggcagtt
     3481 aactagtgca ctgcagtaca aacacagtcc tttcccgcaa ttttcttttt ctattactct
     3541 tggcctcctc tagtacactc tatatttttt tatgcctcgg taatgatttt catttttttt
     3601 tttccaccta gcggatgact cttttttttt cttagcgatt ggcattatca cataatgaat
     3661 tatacattat ataaagtaat gtgatttctt cgaagaatat actaaaaaat gagcaggcaa
     3721 gataaacgaa ggcaaagatg acagagcaga aagccctagt aaagcgtatt acaaatgaaa
     3781 ccaagattca gattgcgatc tctttaaagg gtggtcccct agcgatagag cactcgatct
     3841 tcccagaaaa agaggcagaa gcagtagcag aacaggccac acaatcgcaa gtgattaacg
     3901 tccacacagg tatagggttt ctggaccata tgatacatgc tctggccaag cattccggct
     3961 ggtcgctaat cgttgagtgc attggtgact tacacataga cgaccatcac accactgagg
     4021 actgcgggat tgctctcggt caagctttta aagaggccct aggggccgtg cgtggagtaa
     4081 aaaggtttgg atcaggattt gcgcctttgg atgaggcact ttccagagcg gtggttgatc
     4141 tttcgaacag gccgtacgca gttgtcgaac ttggtttgca aagggagaaa gtaggtgatc
     4201 tctcttgcga gatgatcccg cattttcttg aaagctttgc agaggctagc agaattaccc
     4261 tccacgttga ttgtctgcga ggcaagaatg atcatcaccg tagtgagagt gcgttcaagg
     4321 ctcttgcggt tgccataaga gaagccacct cgcccaatgg taccaacgat gttccctcca
     4381 ccaaaggtgt tcttatgtag tgacaccgat tatttaaagc tgctgcatac gatatatata
     4441 catgtgtata tatgtatacc tatgaatgtc agtaagtatg tatacgaaca gtatgatact
     4501 gaagatgaca aggtaatgca tcattctata cgtgtcattc tgaacgaggc gcgctttcct
     4561 tttttctttt tgctttttct ttttttttct cttgaactcg acggatcata gagtatcacg
     4621 tgctataaaa ataattataa tttaaatttt ttaatataaa tatataaatt aaaaatagaa
     4681 agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc cgaagatgta aaagactcta
     4741 gggggatcgc caacaaatac taccttttat cttgctcttc ctgctctcag gtattaatgc
     4801 cgaattgttt catcttgtct gtgtagaaaa ccacacacga aaatcctgtg attttacatt
     4861 ttacttatcg ttaatcgaat gtatatctat ttaatctgct tttcttgtct aataaatata
     4921 tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt gtttattttt ttttcttcat
     4981 tccgtaactc ttctaccttc tttatttact ttctaaaatc caaatacaaa acataaaaat
     5041 aaataaacac agagtaaatt cccaaattat tccatcatta aaagatacga ggcgcgtgta
     5101 agttacaggc aagcgatccg tcccgagcgg ccgcgattat caaaaaggat cttcacctag
     5161 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5221 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5281 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5341 tctggcccca gtgctgcaat gataccgcgg gacccacgct caccggctcc agatttatca
     5401 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5461 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5521 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5581 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5641 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5701 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5761 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5821 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5881 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5941 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6001 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6061 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6121 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6181 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg tcatgaccaa aatcccttaa
     6241 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6301 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6361 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6421 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6481 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6541 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6601 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6661 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6721 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6781 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6841 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6901 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6961 tcccctgatt ctgtggataa ccgtgcggcc gc
//
