LOCUS       sequence-12731-e        3993 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 3993)
  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 3993)
  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-HTB-TRP1
FEATURES             Location/Qualifiers
     source          1..3993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             72..89
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             117..137
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     terminator      431..618
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(486..510)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        654..801
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             802..1476
                     /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             1550..1558
                     /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(1630..1649)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1631..1649)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1736..1753)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1907..2495)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1987..2006)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2666..3457,3458..3526))
                     /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     3289..3308
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3527..3631)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3699..3717
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3755..3777)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3877..3896
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(3977..3993,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(3977..3993,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acaggggttc acatcatcac caccatcatg actacgatat acccacaacc gccagcgaga
      121 atctgtactt tcaatcagag ttaaaaaccg ccgcactagc tcaacatgac gaggctgctg
      181 gaaaggccgg tgaaggtgaa atccctgccc ctcttgctgg taccgtttct aagatactgg
      241 taaaagaagg tgacactgtt aaagctggtc aaacagttct ggtgctggag gctatgaaaa
      301 tggagacaga aattaacgct cctactgacg gaaaagttga aaaggtgtta gttaaggaaa
      361 gagatgctgt tcaaggtggt caaggtctaa tcaagatcgg cgtttaatga ggcgcgccac
      421 ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa
      481 aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt
      541 gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg tcgctcttat
      601 tgaccacacc tctaccggca gatccgctag ggataacagg gtaatataga tctgtacaat
      661 cttgatccgg agcttttctt tttttgccga ttaagaatta attcggtcga aaaaagaaaa
      721 ggagagggcc aagagggagg gcattggtga ctattgagca cgtgagtata cgtgattaag
      781 cacacaaagg cagcttggag tatgtctgtt attaatttca caggtagttc tggtccattg
      841 gtgaaagttt gcggcttgca gagcacagag gccgcagaat gtgctctaga ttccgatgct
      901 gacttgctgg gtattatatg tgtgcccaat agaaagagaa caattgaccc ggttattgca
      961 aggaaaattt caagtcttgt aaaagcatat aaaaatagtt caggcactcc gaaatacttg
     1021 gttggcgtgt ttcgtaatca acctaaggag gatgttttgg ctctggtcaa tgattacggc
     1081 attgatatcg tccaactgca tggagatgag tcgtggcaag aataccaaga gttcctcggt
     1141 ttgccagtta ttaaaagact cgtatttcca aaagactgca acatactact cagtgcagct
     1201 tcacagaaac ctcattcgtt tattcccttg tttgattcag aagcaggtgg gacaggtgaa
     1261 cttttggatt ggaactcgat ttctgactgg gttggaaggc aagagagccc cgaaagctta
     1321 cattttatgt tagctggtgg actgacgcca gaaaatgttg gtgatgcgct tagattaaat
     1381 ggcgttattg gtgttgatgt aagcggaggt gtggagacaa atggtgtaaa agactctaac
     1441 aaaatagcaa atttcgtcaa aaatgctaag aaataggtta ttactgagta gtatttattt
     1501 aagtattgtt tgtgcacttg cctgcggtgt gaaataccgc acagatgcgt aaggaggttt
     1561 aaacgagctc gaattcatcg atgatatcag atccactagt ggcctatgcg gccgcggatc
     1621 tgccggtctc cctatagtga gtcgtattaa tttcgataag ccaggttaac ctgcattaat
     1681 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     1741 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1801 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1861 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1921 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1981 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2041 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2101 aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2161 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2221 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2281 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2341 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2401 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2461 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2521 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2581 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2641 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2701 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     2761 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     2821 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     2881 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     2941 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3001 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3061 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3121 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3181 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     3241 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     3301 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     3361 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     3421 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     3481 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     3541 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3601 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     3661 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     3721 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     3781 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     3841 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     3901 gagtgcacca tatggacata ttgtcgttag aacgcggcta caattaatac ataaccttat
     3961 gtatcataca catacgattt aggtgacact ata
//
