LOCUS       sequence-9669-ex        4115 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 4115)
  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 4115)
  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-V5-HIS3MX6
FEATURES             Location/Qualifiers
     source          1..4115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             76..117
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     terminator      141..328
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(196..220)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        403..746
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             747..1445
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSNRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1449..1646
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1750..1769)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1751..1769)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1856..1873)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2027..2615)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2107..2126)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2786..3577,3578..3646))
                     /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     3409..3428
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3647..3751)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3819..3837
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3875..3897)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3997..4016
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4097..4115
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4097..4114
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gggggaggcg ggggtggaaa gcctatccct aaccctctcc tcggtctcga ttctacgtag
      121 ggcgcgccac ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag
      181 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat
      241 tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg
      301 tcgctcttat tgaccacacc tctaccggca gatccgctag ggataacagg gtaatataga
      361 tctgtttagc ttgcctcgtc cccgccgggt cacccggcca gcgacatgga ggcccagaat
      421 accctccttg acagtcttga cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac
      481 atttagccca tacatcccca tgtataatca tttgcatcca tacattttga tggccgcacg
      541 gcgcgaagca aaaattacgg ctcctcgctg cagacctgcg agcagggaaa cgctcccctc
      601 acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat
      661 ttgccactga ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct
      721 cacatcacat ccgaacataa acaaccatgg cagaaccagc ccaaaaaaag caaaaacaaa
      781 ctgttcagga gcgcaaggcg tttatctccc gtatcactaa tgaaactaaa attcaaatcg
      841 ctatttcgct gaatggtggt tatattcaaa taaaagattc gattcttcct gcaaagaagg
      901 atgacgatgt agcttcccaa gctactcagt cacaggtcat cgatattcac acaggtgttg
      961 gctttttgga tcatatgatc catgcgttgg caaaacactc tggttggtct cttattgttg
     1021 aatgtatcgg tgacctgcac attgacgatc accatactac cgaagattgc ggtatcgcat
     1081 tagggcaagc gttcaaagaa gcaatgggtg ctgtccgtgg tgtaaaaaga ttcggtactg
     1141 ggttcgcacc attggatgag gcgctatcac gtgccgtagt cgatttatct aatagaccat
     1201 ttgctgtaat cgaccttgga ttgaagagag agatgattgg tgatttatcc actgaaatga
     1261 ttccacactt tttggaaagt ttcgcggagg cggccagaat tactttgcat gttgattgtc
     1321 tgagaggttt caacgatcac cacagaagtg agagtgcgtt caaggctttg gctgttgcca
     1381 taagagaagc tatttctagc aatggcacca atgacgttcc ctcaaccaaa ggtgttttga
     1441 tgtgaagtac tgacaataaa aagattcttg ttttcaagaa cttgtcattt gtatagtttt
     1501 tttatattgt agttgttcta ttttaatcaa atgttagcgt gatttatatt ttttttcgcc
     1561 tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc
     1621 gtatgtgaat gctggtcgct atactgctgt cgattcgata ctaacgccgc catccagttt
     1681 aaacgagctc gaattcatcg atgatatcag atccactagt ggcctatgcg gccgcggatc
     1741 tgccggtctc cctatagtga gtcgtattaa tttcgataag ccaggttaac ctgcattaat
     1801 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     1861 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1921 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1981 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2041 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2101 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2161 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2221 aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2281 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2341 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2401 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2461 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2521 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2581 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2641 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2701 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2761 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2821 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     2881 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     2941 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3001 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3061 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3121 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3181 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3241 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3301 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     3361 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     3421 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     3481 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     3541 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     3601 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     3661 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3721 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     3781 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     3841 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     3901 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     3961 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     4021 gagtgcacca tatggacata ttgtcgttag aacgcggcta caattaatac ataaccttat
     4081 gtatcataca catacgattt aggtgacact ataga
//
