LOCUS       sequence-55638-e        4730 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 4730)
  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 4730)
  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-His3MX6
FEATURES             Location/Qualifiers
     source          1..4730
                     /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        1018..1361
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1362..2060
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSNRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      2064..2261
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2365..2384)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2366..2384)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2471..2488)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2642..3230)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2722..2741)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3401..4192,4193..4261))
                     /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     4024..4043
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4262..4366)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4434..4452
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4490..4512)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4612..4631
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4712..4730
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4712..4729
                     /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 ttagcttgcc tcgtccccgc cgggtcaccc ggccagcgac
     1021 atggaggccc agaataccct ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt
     1081 gactgtcgcc cgtacattta gcccatacat ccccatgtat aatcatttgc atccatacat
     1141 tttgatggcc gcacggcgcg aagcaaaaat tacggctcct cgctgcagac ctgcgagcag
     1201 ggaaacgctc ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat
     1261 ataaaaggtt aggatttgcc actgaggttc ttctttcata tacttccttt taaaatcttg
     1321 ctaggataca gttctcacat cacatccgaa cataaacaac catggcagaa ccagcccaaa
     1381 aaaagcaaaa acaaactgtt caggagcgca aggcgtttat ctcccgtatc actaatgaaa
     1441 ctaaaattca aatcgctatt tcgctgaatg gtggttatat tcaaataaaa gattcgattc
     1501 ttcctgcaaa gaaggatgac gatgtagctt cccaagctac tcagtcacag gtcatcgata
     1561 ttcacacagg tgttggcttt ttggatcata tgatccatgc gttggcaaaa cactctggtt
     1621 ggtctcttat tgttgaatgt atcggtgacc tgcacattga cgatcaccat actaccgaag
     1681 attgcggtat cgcattaggg caagcgttca aagaagcaat gggtgctgtc cgtggtgtaa
     1741 aaagattcgg tactgggttc gcaccattgg atgaggcgct atcacgtgcc gtagtcgatt
     1801 tatctaatag accatttgct gtaatcgacc ttggattgaa gagagagatg attggtgatt
     1861 tatccactga aatgattcca cactttttgg aaagtttcgc ggaggcggcc agaattactt
     1921 tgcatgttga ttgtctgaga ggtttcaacg atcaccacag aagtgagagt gcgttcaagg
     1981 ctttggctgt tgccataaga gaagctattt ctagcaatgg caccaatgac gttccctcaa
     2041 ccaaaggtgt tttgatgtga agtactgaca ataaaaagat tcttgttttc aagaacttgt
     2101 catttgtata gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt
     2161 atattttttt tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa
     2221 tatcatgcgt caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac
     2281 gccgccatcc agtttaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct
     2341 atgcggccgc ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg
     2401 ttaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2461 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2521 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2581 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2641 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2701 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2761 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2821 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2881 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2941 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3001 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3061 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     3121 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3181 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3241 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3301 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3361 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3421 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3481 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3541 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3601 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3661 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3721 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3781 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3841 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3901 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3961 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4021 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4081 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4141 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4201 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4261 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4321 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4381 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     4441 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     4501 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     4561 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     4621 agcagattgt actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt
     4681 aatacataac cttatgtatc atacacatac gatttaggtg acactataga
//
