LOCUS       sequence-9642-ex        4100 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 4100)
  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 4100)
  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-FLAG-HIS3MX6
FEATURES             Location/Qualifiers
     source          1..4100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             79..102
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      126..313
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(181..205)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        388..731
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             732..1430
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSNRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1434..1631
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1735..1754)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1736..1754)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1841..1858)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2012..2600)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2092..2111)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2771..3562,3563..3631))
                     /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     3394..3413
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3632..3736)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3804..3822
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3860..3882)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3982..4001
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4082..4100
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4082..4099
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gggggaggcg ggggtggaga ttacaaggac gacgatgaca agtagggcgc gccacttcta
      121 aataagcgaa tttcttatga tttatgattt ttattattaa ataagttata aaaaaaataa
      181 gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta ttcttgagta
      241 actctttcct gtaggtcagg ttgctttctc aggtatagta tgaggtcgct cttattgacc
      301 acacctctac cggcagatcc gctagggata acagggtaat atagatctgt ttagcttgcc
      361 tcgtccccgc cgggtcaccc ggccagcgac atggaggccc agaataccct ccttgacagt
      421 cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc cgtacattta gcccatacat
      481 ccccatgtat aatcatttgc atccatacat tttgatggcc gcacggcgcg aagcaaaaat
      541 tacggctcct cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat
      601 tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc actgaggttc
      661 ttctttcata tacttccttt taaaatcttg ctaggataca gttctcacat cacatccgaa
      721 cataaacaac catggcagaa ccagcccaaa aaaagcaaaa acaaactgtt caggagcgca
      781 aggcgtttat ctcccgtatc actaatgaaa ctaaaattca aatcgctatt tcgctgaatg
      841 gtggttatat tcaaataaaa gattcgattc ttcctgcaaa gaaggatgac gatgtagctt
      901 cccaagctac tcagtcacag gtcatcgata ttcacacagg tgttggcttt ttggatcata
      961 tgatccatgc gttggcaaaa cactctggtt ggtctcttat tgttgaatgt atcggtgacc
     1021 tgcacattga cgatcaccat actaccgaag attgcggtat cgcattaggg caagcgttca
     1081 aagaagcaat gggtgctgtc cgtggtgtaa aaagattcgg tactgggttc gcaccattgg
     1141 atgaggcgct atcacgtgcc gtagtcgatt tatctaatag accatttgct gtaatcgacc
     1201 ttggattgaa gagagagatg attggtgatt tatccactga aatgattcca cactttttgg
     1261 aaagtttcgc ggaggcggcc agaattactt tgcatgttga ttgtctgaga ggtttcaacg
     1321 atcaccacag aagtgagagt gcgttcaagg ctttggctgt tgccataaga gaagctattt
     1381 ctagcaatgg caccaatgac gttccctcaa ccaaaggtgt tttgatgtga agtactgaca
     1441 ataaaaagat tcttgttttc aagaacttgt catttgtata gtttttttat attgtagttg
     1501 ttctatttta atcaaatgtt agcgtgattt atattttttt tcgcctcgac atcatctgcc
     1561 cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg
     1621 tcgctatact gctgtcgatt cgatactaac gccgccatcc agtttaaacg agctcgaatt
     1681 catcgatgat atcagatcca ctagtggcct atgcggccgc ggatctgccg gtctccctat
     1741 agtgagtcgt attaatttcg ataagccagg ttaacctgca ttaatgaatc ggccaacgcg
     1801 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     1861 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1921 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     1981 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2041 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2101 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2161 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     2221 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2281 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2341 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2401 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     2461 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2521 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2581 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2641 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2701 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2761 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2821 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     2881 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     2941 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3001 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3061 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3121 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3181 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3241 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3301 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3361 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3421 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3481 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3541 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3601 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     3661 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     3721 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     3781 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     3841 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     3901 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     3961 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgg
     4021 acatattgtc gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac
     4081 gatttaggtg acactataga
//
