LOCUS       sequence-55632-e        3753 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 3753)
  AUTHORS   Longtine MS, McKenzie A 3rd, Demarini DJ, Shah NG, Wach A, Brachat 
            A, Philippsen P, Pringle JR
  TITLE     Additional modules for versatile and economical PCR-based gene 
            deletion and modification in Saccharomyces cerevisiae.
  JOURNAL   Yeast. 1998 Jul;14(10):953-61.
  PUBMED    9717241
REFERENCE   2  (bases 1 to 3753)
  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."; 
            date: "1998-07"; volume: "14(10)"; pages: "953-61"
COMMENT     Sequence Label: pFA6a-3HA-TRP1
FEATURES             Location/Qualifiers
     source          1..3753
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             67..156
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     terminator      189..376
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(244..268)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        412..559
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             560..1234
                     /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             1308..1316
                     /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(1388..1407)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1389..1407)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1494..1511)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1665..2253)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1745..1764)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2424..3215,3216..3284))
                     /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     3047..3066
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3285..3389)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3457..3475
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3513..3535)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3635..3654
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3735..3753
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3735..3752
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 atcttttacc catacgatgt tcctgactat gcgggctatc cgtatgacgt cccggactat
      121 gcaggatcct atccatatga cgttccagat tacgctgctc agtgctgagg cgcgccactt
      181 ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa
      241 taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga
      301 gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc gctcttattg
      361 accacacctc taccggcaga tccgctaggg ataacagggt aatatagatc tgtacaatct
      421 tgatccggag cttttctttt tttgccgatt aagaattaat tcggtcgaaa aaagaaaagg
      481 agagggccaa gagggagggc attggtgact attgagcacg tgagtatacg tgattaagca
      541 cacaaaggca gcttggagta tgtctgttat taatttcaca ggtagttctg gtccattggt
      601 gaaagtttgc ggcttgcaga gcacagaggc cgcagaatgt gctctagatt ccgatgctga
      661 cttgctgggt attatatgtg tgcccaatag aaagagaaca attgacccgg ttattgcaag
      721 gaaaatttca agtcttgtaa aagcatataa aaatagttca ggcactccga aatacttggt
      781 tggcgtgttt cgtaatcaac ctaaggagga tgttttggct ctggtcaatg attacggcat
      841 tgatatcgtc caactgcatg gagatgagtc gtggcaagaa taccaagagt tcctcggttt
      901 gccagttatt aaaagactcg tatttccaaa agactgcaac atactactca gtgcagcttc
      961 acagaaacct cattcgttta ttcccttgtt tgattcagaa gcaggtggga caggtgaact
     1021 tttggattgg aactcgattt ctgactgggt tggaaggcaa gagagccccg aaagcttaca
     1081 ttttatgtta gctggtggac tgacgccaga aaatgttggt gatgcgctta gattaaatgg
     1141 cgttattggt gttgatgtaa gcggaggtgt ggagacaaat ggtgtaaaag actctaacaa
     1201 aatagcaaat ttcgtcaaaa atgctaagaa ataggttatt actgagtagt atttatttaa
     1261 gtattgtttg tgcacttgcc tgcggtgtga aataccgcac agatgcgtaa ggaggtttaa
     1321 acgagctcga attcatcgat gatatcagat ccactagtgg cctatgcggc cgcggatctg
     1381 ccggtctccc tatagtgagt cgtattaatt tcgataagcc aggttaacct gcattaatga
     1441 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1501 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1561 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1621 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1681 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1741 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     1801 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     1861 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     1921 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     1981 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2041 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2101 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2161 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2221 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2281 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2341 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2401 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2461 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2521 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2581 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2641 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2701 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     2761 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     2821 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     2881 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     2941 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3001 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3061 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3121 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3181 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3241 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3301 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3361 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     3421 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     3481 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     3541 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     3601 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     3661 gtgcaccata tggacatatt gtcgttagaa cgcggctaca attaatacat aaccttatgt
     3721 atcatacaca tacgatttag gtgacactat aga
//
