LOCUS       sequence-9654-ex        4094 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 4094)
  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 4094)
  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-His-tag-HIS3MX6
FEATURES             Location/Qualifiers
     source          1..4094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             79..96
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      120..307
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(175..199)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        382..725
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             726..1424
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSNRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1428..1625
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1729..1748)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1730..1748)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1835..1852)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2006..2594)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2086..2105)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2765..3556,3557..3625))
                     /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     3388..3407
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3626..3730)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3798..3816
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3854..3876)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3976..3995
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4076..4094
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4076..4093
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gggggaggcg ggggtggaca tcatcaccat caccactagg gcgcgccact tctaaataag
      121 cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat
      181 acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt
      241 tcctgtaggt caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct
      301 ctaccggcag atccgctagg gataacaggg taatatagat ctgtttagct tgcctcgtcc
      361 ccgccgggtc acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac
      421 gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat
      481 gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc
      541 tcctcgctgc agacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc
      601 cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt
      661 catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa
      721 caaccatggc agaaccagcc caaaaaaagc aaaaacaaac tgttcaggag cgcaaggcgt
      781 ttatctcccg tatcactaat gaaactaaaa ttcaaatcgc tatttcgctg aatggtggtt
      841 atattcaaat aaaagattcg attcttcctg caaagaagga tgacgatgta gcttcccaag
      901 ctactcagtc acaggtcatc gatattcaca caggtgttgg ctttttggat catatgatcc
      961 atgcgttggc aaaacactct ggttggtctc ttattgttga atgtatcggt gacctgcaca
     1021 ttgacgatca ccatactacc gaagattgcg gtatcgcatt agggcaagcg ttcaaagaag
     1081 caatgggtgc tgtccgtggt gtaaaaagat tcggtactgg gttcgcacca ttggatgagg
     1141 cgctatcacg tgccgtagtc gatttatcta atagaccatt tgctgtaatc gaccttggat
     1201 tgaagagaga gatgattggt gatttatcca ctgaaatgat tccacacttt ttggaaagtt
     1261 tcgcggaggc ggccagaatt actttgcatg ttgattgtct gagaggtttc aacgatcacc
     1321 acagaagtga gagtgcgttc aaggctttgg ctgttgccat aagagaagct atttctagca
     1381 atggcaccaa tgacgttccc tcaaccaaag gtgttttgat gtgaagtact gacaataaaa
     1441 agattcttgt tttcaagaac ttgtcatttg tatagttttt ttatattgta gttgttctat
     1501 tttaatcaaa tgttagcgtg atttatattt tttttcgcct cgacatcatc tgcccagatg
     1561 cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta
     1621 tactgctgtc gattcgatac taacgccgcc atccagttta aacgagctcg aattcatcga
     1681 tgatatcaga tccactagtg gcctatgcgg ccgcggatct gccggtctcc ctatagtgag
     1741 tcgtattaat ttcgataagc caggttaacc tgcattaatg aatcggccaa cgcgcgggga
     1801 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     1861 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1921 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1981 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2041 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2101 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2161 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     2221 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2281 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2341 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2401 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     2461 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2521 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2581 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2641 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     2701 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     2761 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     2821 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     2881 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     2941 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     3001 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     3061 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     3121 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     3181 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     3241 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     3301 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     3361 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     3421 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     3481 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     3541 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     3601 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     3661 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     3721 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     3781 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     3841 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     3901 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     3961 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atggacatat
     4021 tgtcgttaga acgcggctac aattaataca taaccttatg tatcatacac atacgattta
     4081 ggtgacacta taga
//
