LOCUS       sequence-55637-e        4320 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4320)
  AUTHORS   Longtine MS, McKenzie A 3rd, Demarini DJ, Shah NG, Wach A, Brachat 
            A, Philippsen P, Pringle JR
  TITLE     Additional modules for versatile and economical PCR-based gene 
            deletion and modification in Saccharomyces cerevisiae.
  JOURNAL   Yeast. 1998 Jul;14(10):953-61.
  PUBMED    9717241
REFERENCE   2  (bases 1 to 4320)
  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: "Yeast."; 
            date: "1998-07"; volume: "14(10)"; pages: "953-61"
COMMENT     Sequence Label: pFA6a-GST-TRP1
FEATURES             Location/Qualifiers
     source          1..4320
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             61..714
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     672..694
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     terminator      756..943
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(811..835)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        979..1126
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             1127..1801
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     RBS             1875..1883
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(1955..1974)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1956..1974)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2061..2078)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2232..2820)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2312..2331)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2991..3782,3783..3851))
                     /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     3614..3633
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3852..3956)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4024..4042
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4080..4102)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4202..4221
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4302..4320
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4302..4319
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaat
       61 atgtccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt
      121 ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa
      181 tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta ttatattgat
      241 ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac
      301 atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg
      361 gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt
      421 gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa
      481 acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat
      541 gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa
      601 aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca
      661 tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc aaaatcggat
      721 ctggttccgc gttgaggcgc gccacttcta aataagcgaa tttcttatga tttatgattt
      781 ttattattaa ataagttata aaaaaaataa gtgtatacaa attttaaagt gactcttagg
      841 ttttaaaacg aaaattctta ttcttgagta actctttcct gtaggtcagg ttgctttctc
      901 aggtatagta tgaggtcgct cttattgacc acacctctac cggcagatcc gctagggata
      961 acagggtaat atagatctgt acaatcttga tccggagctt ttcttttttt gccgattaag
     1021 aattaattcg gtcgaaaaaa gaaaaggaga gggccaagag ggagggcatt ggtgactatt
     1081 gagcacgtga gtatacgtga ttaagcacac aaaggcagct tggagtatgt ctgttattaa
     1141 tttcacaggt agttctggtc cattggtgaa agtttgcggc ttgcagagca cagaggccgc
     1201 agaatgtgct ctagattccg atgctgactt gctgggtatt atatgtgtgc ccaatagaaa
     1261 gagaacaatt gacccggtta ttgcaaggaa aatttcaagt cttgtaaaag catataaaaa
     1321 tagttcaggc actccgaaat acttggttgg cgtgtttcgt aatcaaccta aggaggatgt
     1381 tttggctctg gtcaatgatt acggcattga tatcgtccaa ctgcatggag atgagtcgtg
     1441 gcaagaatac caagagttcc tcggtttgcc agttattaaa agactcgtat ttccaaaaga
     1501 ctgcaacata ctactcagtg cagcttcaca gaaacctcat tcgtttattc ccttgtttga
     1561 ttcagaagca ggtgggacag gtgaactttt ggattggaac tcgatttctg actgggttgg
     1621 aaggcaagag agccccgaaa gcttacattt tatgttagct ggtggactga cgccagaaaa
     1681 tgttggtgat gcgcttagat taaatggcgt tattggtgtt gatgtaagcg gaggtgtgga
     1741 gacaaatggt gtaaaagact ctaacaaaat agcaaatttc gtcaaaaatg ctaagaaata
     1801 ggttattact gagtagtatt tatttaagta ttgtttgtgc acttgcctgc ggtgtgaaat
     1861 accgcacaga tgcgtaagga ggtttaaacg agctcgaatt catcgatgat atcagatcca
     1921 ctagtggcct atgcggccgc ggatctgccg gtctccctat agtgagtcgt attaatttcg
     1981 ataagccagg ttaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     2041 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2101 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2161 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2221 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2281 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2341 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2401 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     2461 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2521 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2581 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2641 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     2701 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2761 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2821 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2881 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     2941 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3001 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3061 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3121 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3181 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3241 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3301 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3361 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3421 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3481 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3541 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     3601 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     3661 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     3721 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     3781 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     3841 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     3901 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     3961 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     4021 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     4081 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     4141 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     4201 gcggcatcag agcagattgt actgagagtg caccatatgg acatattgtc gttagaacgc
     4261 ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg acactataga
//
