LOCUS       sequence-9661-ex        4219 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 4219)
  AUTHORS   Funakoshi M, Hochstrasser M
  TITLE     Small epitope-linker modules for PCR-based C-terminal tagging in 
            Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2009 Feb 25. 26(3):185-192.
  PUBMED    19243080
REFERENCE   2  (bases 1 to 4219)
  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. 2009
            Feb 25. 26(3):185-192."
COMMENT     Sequence Label: pFA6a-6xGLY-Myc-kanMX6
FEATURES             Location/Qualifiers
     source          1..4219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             79..108
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      132..319
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(187..211)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            394..1750
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        394..737
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             738..1547
                     /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(805..824)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1553..1750
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1854..1873)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1855..1873)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1960..1977)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2131..2719)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2211..2230)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2890..3681,3682..3750))
                     /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     3513..3532
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3751..3855)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3923..3941
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3979..4001)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4101..4120
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4201..4219
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4201..4218
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gggggaggcg ggggtggaga gcagaaactc atctcagaag aggatctgta gggcgcgcca
      121 cttctaaata agcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa
      181 aaataagtgt atacaaattt taaagtgact cttaggtttt aaaacgaaaa ttcttattct
      241 tgagtaactc tttcctgtag gtcaggttgc tttctcaggt atagtatgag gtcgctctta
      301 ttgaccacac ctctaccggc agatccgcta gggataacag ggtaatatag atctgtttag
      361 cttgcctcgt ccccgccggg tcacccggcc agcgacatgg aggcccagaa taccctcctt
      421 gacagtcttg acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta catttagccc
      481 atacatcccc atgtataatc atttgcatcc atacattttg atggccgcac ggcgcgaagc
      541 aaaaattacg gctcctcgct gcagacctgc gagcagggaa acgctcccct cacagacgcg
      601 ttgaattgtc cccacgccgc gcccctgtag agaaatataa aaggttagga tttgccactg
      661 aggttcttct ttcatatact tccttttaaa atcttgctag gatacagttc tcacatcaca
      721 tccgaacata aacaaccatg ggtaaggaaa agactcacgt ttcgaggccg cgattaaatt
      781 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
      841 gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
      901 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
      961 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     1021 tcaccactgc gatccccggc aaaacagcat tccaggtatt agaagaatat cctgattcag
     1081 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     1141 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga
     1201 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     1261 aagtctggaa agaaatgcat aagcttttgc cattctcacc ggattcagtc gtcactcatg
     1321 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     1381 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     1441 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     1501 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gtactgacaa
     1561 taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt
     1621 tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc
     1681 agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt
     1741 cgctatactg ctgtcgattc gatactaacg ccgccatcca gtttaaacga gctcgaattc
     1801 atcgatgata tcagatccac tagtggccta tgcggccgcg gatctgccgg tctccctata
     1861 gtgagtcgta ttaatttcga taagccaggt taacctgcat taatgaatcg gccaacgcgc
     1921 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     1981 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2041 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2101 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2161 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2221 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2281 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     2341 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2401 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2461 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2521 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     2581 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2641 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2701 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2761 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2821 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2881 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2941 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3001 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3061 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3121 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3181 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3241 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3301 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3361 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3421 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3481 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3541 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     3601 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     3661 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3721 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3781 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3841 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     3901 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     3961 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     4021 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     4081 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgga
     4141 catattgtcg ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg
     4201 atttaggtga cactataga
//
