LOCUS       sequence-12738-e        3714 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 3714)
  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 3714)
  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-RGS18HIS-TRP1
FEATURES             Location/Qualifiers
     source          1..3714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             72..101
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             75..104
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             78..107
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             81..110
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             84..113
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             87..116
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             90..119
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             93..122
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             96..125
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     terminator      152..339
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(207..231)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        375..522
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             523..1197
                     /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             1271..1279
                     /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(1351..1370)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1352..1370)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1457..1474)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1628..2216)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1708..1727)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2387..3178,3179..3247))
                     /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     3010..3029
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3248..3352)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3420..3438
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3476..3498)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3598..3617
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(3698..3714,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(3698..3714,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acaggggttc acatcatcac caccatcatc atcatcacca ccatcatcat catcaccacc
      121 atcattaatg aggcgcgcca cttctaaata agcgaatttc ttatgattta tgatttttat
      181 tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact cttaggtttt
      241 aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc tttctcaggt
      301 atagtatgag gtcgctctta ttgaccacac ctctaccggc agatccgcta gggataacag
      361 ggtaatatag atctgtacaa tcttgatccg gagcttttct ttttttgccg attaagaatt
      421 aattcggtcg aaaaaagaaa aggagagggc caagagggag ggcattggtg actattgagc
      481 acgtgagtat acgtgattaa gcacacaaag gcagcttgga gtatgtctgt tattaatttc
      541 acaggtagtt ctggtccatt ggtgaaagtt tgcggcttgc agagcacaga ggccgcagaa
      601 tgtgctctag attccgatgc tgacttgctg ggtattatat gtgtgcccaa tagaaagaga
      661 acaattgacc cggttattgc aaggaaaatt tcaagtcttg taaaagcata taaaaatagt
      721 tcaggcactc cgaaatactt ggttggcgtg tttcgtaatc aacctaagga ggatgttttg
      781 gctctggtca atgattacgg cattgatatc gtccaactgc atggagatga gtcgtggcaa
      841 gaataccaag agttcctcgg tttgccagtt attaaaagac tcgtatttcc aaaagactgc
      901 aacatactac tcagtgcagc ttcacagaaa cctcattcgt ttattccctt gtttgattca
      961 gaagcaggtg ggacaggtga acttttggat tggaactcga tttctgactg ggttggaagg
     1021 caagagagcc ccgaaagctt acattttatg ttagctggtg gactgacgcc agaaaatgtt
     1081 ggtgatgcgc ttagattaaa tggcgttatt ggtgttgatg taagcggagg tgtggagaca
     1141 aatggtgtaa aagactctaa caaaatagca aatttcgtca aaaatgctaa gaaataggtt
     1201 attactgagt agtatttatt taagtattgt ttgtgcactt gcctgcggtg tgaaataccg
     1261 cacagatgcg taaggaggtt taaacgagct cgaattcatc gatgatatca gatccactag
     1321 tggcctatgc ggccgcggat ctgccggtct ccctatagtg agtcgtatta atttcgataa
     1381 gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     1441 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     1501 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     1561 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1621 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1681 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1741 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1801 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg
     1861 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1921 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1981 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2041 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     2101 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     2161 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     2221 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     2281 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     2341 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     2401 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     2461 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     2521 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     2581 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     2641 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     2701 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     2761 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     2821 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     2881 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     2941 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     3001 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     3061 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     3121 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     3181 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     3241 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     3301 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     3361 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     3421 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     3481 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     3541 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     3601 catcagagca gattgtactg agagtgcacc atatggacat attgtcgtta gaacgcggct
     3661 acaattaata cataacctta tgtatcatac acatacgatt taggtgacac tata
//
