LOCUS       sequence-12728-e        3924 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 3924)
  AUTHORS   Tagwerker C, Zhang H, Wang X, Larsen LS, Lathrop RH, Hatfield GW, 
            Auer B, Huang L, Kaiser P
  TITLE     HB tag modules for PCR-based gene tagging and tandem affinity 
            purification in Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2006 Jun . 23(8):623-32.
  PUBMED    16823883
REFERENCE   2  (bases 1 to 3924)
  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. 2006
            Jun . 23(8):623-32."
COMMENT     Sequence Label: pFA6a-HBH-TRP1
FEATURES             Location/Qualifiers
     source          1..3924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             72..89
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             318..335
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      362..549
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(417..441)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        585..732
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             733..1407
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     RBS             1481..1489
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(1561..1580)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1562..1580)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1667..1684)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1838..2426)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1918..1937)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2597..3388,3389..3457))
                     /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     3220..3239
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3458..3562)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3630..3648
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3686..3708)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3808..3827
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(3908..3924,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(3908..3924,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acaggggttc acatcatcac caccatcatg ctggaaaggc cggtgaaggt gaaatccctg
      121 cccctcttgc tggtaccgtt tctaagatac tggtaaaaga aggtgacact gttaaagctg
      181 gtcaaacagt tctggtgctg gaggctatga aaatggagac agaaattaac gctcctactg
      241 acggaaaagt tgaaaaggtg ttagttaagg aaagagatgc tgttcaaggt ggtcaaggtc
      301 taatcaagat cggcgttcat catcaccacc atcattaatg aggcgcgcca cttctaaata
      361 agcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt
      421 atacaaattt taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc
      481 tttcctgtag gtcaggttgc tttctcaggt atagtatgag gtcgctctta ttgaccacac
      541 ctctaccggc agatccgcta gggataacag ggtaatatag atctgtacaa tcttgatccg
      601 gagcttttct ttttttgccg attaagaatt aattcggtcg aaaaaagaaa aggagagggc
      661 caagagggag ggcattggtg actattgagc acgtgagtat acgtgattaa gcacacaaag
      721 gcagcttgga gtatgtctgt tattaatttc acaggtagtt ctggtccatt ggtgaaagtt
      781 tgcggcttgc agagcacaga ggccgcagaa tgtgctctag attccgatgc tgacttgctg
      841 ggtattatat gtgtgcccaa tagaaagaga acaattgacc cggttattgc aaggaaaatt
      901 tcaagtcttg taaaagcata taaaaatagt tcaggcactc cgaaatactt ggttggcgtg
      961 tttcgtaatc aacctaagga ggatgttttg gctctggtca atgattacgg cattgatatc
     1021 gtccaactgc atggagatga gtcgtggcaa gaataccaag agttcctcgg tttgccagtt
     1081 attaaaagac tcgtatttcc aaaagactgc aacatactac tcagtgcagc ttcacagaaa
     1141 cctcattcgt ttattccctt gtttgattca gaagcaggtg ggacaggtga acttttggat
     1201 tggaactcga tttctgactg ggttggaagg caagagagcc ccgaaagctt acattttatg
     1261 ttagctggtg gactgacgcc agaaaatgtt ggtgatgcgc ttagattaaa tggcgttatt
     1321 ggtgttgatg taagcggagg tgtggagaca aatggtgtaa aagactctaa caaaatagca
     1381 aatttcgtca aaaatgctaa gaaataggtt attactgagt agtatttatt taagtattgt
     1441 ttgtgcactt gcctgcggtg tgaaataccg cacagatgcg taaggaggtt taaacgagct
     1501 cgaattcatc gatgatatca gatccactag tggcctatgc ggccgcggat ctgccggtct
     1561 ccctatagtg agtcgtatta atttcgataa gccaggttaa cctgcattaa tgaatcggcc
     1621 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     1681 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     1741 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     1801 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     1861 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     1921 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1981 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
     2041 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2101 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2161 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     2221 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     2281 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     2341 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     2401 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2461 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     2521 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     2581 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     2641 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     2701 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     2761 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     2821 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     2881 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     2941 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3001 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     3061 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     3121 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     3181 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     3241 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     3301 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     3361 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3421 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     3481 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     3541 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     3601 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     3661 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     3721 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     3781 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     3841 atatggacat attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac
     3901 acatacgatt taggtgacac tata
//
