LOCUS       sequence-49549-e        4843 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 4843)
  AUTHORS   Bahler J, Wu JQ, Longtine MS, Shah NG, McKenzie A 3rd, Steever AB, 
            Wach A, Philippsen P, Pringle JR
  TITLE     Heterologous modules for efficient and versatile PCR-based gene 
            targeting in Schizosaccharomyces pombe.
  JOURNAL   Yeast. 1998 Jul;14(10):943-51.
  PUBMED    9717240
REFERENCE   2  (bases 1 to 4843)
  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: "943-51"
COMMENT     Sequence Label: pFA6a-GST-kanMX6
FEATURES             Location/Qualifiers
     source          1..4843
                     /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"
     gene            1018..2374
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1018..1361
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1362..2171
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1429..1448)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2177..2374
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2478..2497)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2479..2497)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2584..2601)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2755..3343)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2835..2854)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3514..4305,4306..4374))
                     /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     4137..4156
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4375..4479)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4547..4565
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4603..4625)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4725..4744
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4825..4843
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4825..4842
                     /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 catgggtaag gaaaagactc
     1381 acgtttcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     1441 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg
     1501 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     1561 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     1621 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggcaaaaca gcattccagg
     1681 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     1741 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     1801 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     1861 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataagctt ttgccattct
     1921 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     1981 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     2041 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     2101 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     2161 agtttttcta atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt
     2221 atagtttttt tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt
     2281 ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg
     2341 cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca
     2401 tccagtttaa acgagctcga attcatcgat gatatcagat ccactagtgg cctatgcggc
     2461 cgcggatctg ccggtctccc tatagtgagt cgtattaatt tcgataagcc aggttaacct
     2521 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2581 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2641 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2701 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2761 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2821 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2881 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2941 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3001 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3061 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3121 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3181 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3241 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3301 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3361 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3421 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3481 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3541 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3601 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3661 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3721 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3781 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3841 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3901 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3961 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4021 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4081 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4141 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4201 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4261 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4321 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4381 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4441 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4501 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     4561 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     4621 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     4681 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     4741 tgtactgaga gtgcaccata tggacatatt gtcgttagaa cgcggctaca attaatacat
     4801 aaccttatgt atcatacaca tacgatttag gtgacactat aga
//
