LOCUS       sequence-9667-ex        4225 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 4225)
  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 4225)
  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-HSV-kanMX6
FEATURES             Location/Qualifiers
     source          1..4225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             82..114
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     terminator      138..325
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(193..217)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            400..1756
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        400..743
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             744..1553
                     /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(811..830)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1559..1756
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1860..1879)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1861..1879)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1966..1983)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2137..2725)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2217..2236)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2896..3687,3688..3756))
                     /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     3519..3538
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3757..3861)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3929..3947
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3985..4007)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4107..4126
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4207..4225
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4207..4224
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gggggaggcg ggggtggaag ccagccagaa ctcgccccgg aagaccccga ggattagggc
      121 gcgccacttc taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta
      181 taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct
      241 tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag tatgaggtcg
      301 ctcttattga ccacacctct accggcagat ccgctaggga taacagggta atatagatct
      361 gtttagcttg cctcgtcccc gccgggtcac ccggccagcg acatggaggc ccagaatacc
      421 ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg cccgtacatt
      481 tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg ccgcacggcg
      541 cgaagcaaaa attacggctc ctcgctgcag acctgcgagc agggaaacgc tcccctcaca
      601 gacgcgttga attgtcccca cgccgcgccc ctgtagagaa atataaaagg ttaggatttg
      661 ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata cagttctcac
      721 atcacatccg aacataaaca accatgggta aggaaaagac tcacgtttcg aggccgcgat
      781 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
      841 aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga
      901 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
      961 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     1021 ggttactcac cactgcgatc cccggcaaaa cagcattcca ggtattagaa gaatatcctg
     1081 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     1141 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
     1201 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     1261 ttgaacaagt ctggaaagaa atgcataagc ttttgccatt ctcaccggat tcagtcgtca
     1321 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     1381 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     1441 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     1501 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagtac
     1561 tgacaataaa aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt
     1621 agttgttcta ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat
     1681 ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat
     1741 gctggtcgct atactgctgt cgattcgata ctaacgccgc catccagttt aaacgagctc
     1801 gaattcatcg atgatatcag atccactagt ggcctatgcg gccgcggatc tgccggtctc
     1861 cctatagtga gtcgtattaa tttcgataag ccaggttaac ctgcattaat gaatcggcca
     1921 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1981 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2041 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2101 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2161 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2221 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2281 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     2341 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2401 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2461 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2521 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     2581 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2641 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2701 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2761 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2821 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2881 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2941 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3001 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3061 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3121 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3181 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3241 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3301 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3361 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3421 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3481 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3541 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3601 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3661 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3721 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3781 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3841 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     3901 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     3961 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     4021 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     4081 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     4141 tatggacata ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca
     4201 catacgattt aggtgacact ataga
//
