LOCUS       sequence-12735-e        3900 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3900)
  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 3900)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-BIO-TRP1
FEATURES             Location/Qualifiers
     source          1..3900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      338..525
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(393..417)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        561..708
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             709..1383
                     /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             1457..1465
                     /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(1537..1556)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1538..1556)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1643..1660)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1814..2402)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1894..1913)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2573..3364,3365..3433))
                     /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     3196..3215
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3434..3538)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3606..3624
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3662..3684)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3784..3803
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(3884..3900,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(3884..3900,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acgctggaaa ggccggtgaa ggtgctggaa aggccggtga aggtgaaatc cctgcccctc
      121 ttgctggtac cgtttctaag atactggtaa aagaaggtga cactgttaaa gctggtcaaa
      181 cagttctggt gctggaggct atgaaaatgg agacagaaat taacgctcct actgacggaa
      241 aagttgaaaa ggtgttagtt aaggaaagag atgctgttca aggtggtcaa ggtctaatca
      301 agatcggcgt ttaatgaggc gcgccacttc taaataagcg aatttcttat gatttatgat
      361 ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta
      421 ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc
      481 tcaggtatag tatgaggtcg ctcttattga ccacacctct accggcagat ccgctaggga
      541 taacagggta atatagatct gtacaatctt gatccggagc ttttcttttt ttgccgatta
      601 agaattaatt cggtcgaaaa aagaaaagga gagggccaag agggagggca ttggtgacta
      661 ttgagcacgt gagtatacgt gattaagcac acaaaggcag cttggagtat gtctgttatt
      721 aatttcacag gtagttctgg tccattggtg aaagtttgcg gcttgcagag cacagaggcc
      781 gcagaatgtg ctctagattc cgatgctgac ttgctgggta ttatatgtgt gcccaataga
      841 aagagaacaa ttgacccggt tattgcaagg aaaatttcaa gtcttgtaaa agcatataaa
      901 aatagttcag gcactccgaa atacttggtt ggcgtgtttc gtaatcaacc taaggaggat
      961 gttttggctc tggtcaatga ttacggcatt gatatcgtcc aactgcatgg agatgagtcg
     1021 tggcaagaat accaagagtt cctcggtttg ccagttatta aaagactcgt atttccaaaa
     1081 gactgcaaca tactactcag tgcagcttca cagaaacctc attcgtttat tcccttgttt
     1141 gattcagaag caggtgggac aggtgaactt ttggattgga actcgatttc tgactgggtt
     1201 ggaaggcaag agagccccga aagcttacat tttatgttag ctggtggact gacgccagaa
     1261 aatgttggtg atgcgcttag attaaatggc gttattggtg ttgatgtaag cggaggtgtg
     1321 gagacaaatg gtgtaaaaga ctctaacaaa atagcaaatt tcgtcaaaaa tgctaagaaa
     1381 taggttatta ctgagtagta tttatttaag tattgtttgt gcacttgcct gcggtgtgaa
     1441 ataccgcaca gatgcgtaag gaggtttaaa cgagctcgaa ttcatcgatg atatcagatc
     1501 cactagtggc ctatgcggcc gcggatctgc cggtctccct atagtgagtc gtattaattt
     1561 cgataagcca ggttaacctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1621 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1681 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1741 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1801 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1861 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1921 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1981 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     2041 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2101 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2161 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2221 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     2281 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2341 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2401 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2461 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2521 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2581 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2641 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     2701 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     2761 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     2821 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     2881 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     2941 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3001 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3061 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3121 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3181 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3241 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3301 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3361 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3421 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3481 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3541 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     3601 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     3661 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     3721 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     3781 atgcggcatc agagcagatt gtactgagag tgcaccatat ggacatattg tcgttagaac
     3841 gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg tgacactata
//
