LOCUS       sequence-55634-e        4163 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 4163)
  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 4163)
  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-His3MX6
FEATURES             Location/Qualifiers
     source          1..4163
                     /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        451..794
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             795..1493
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSNRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1497..1694
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1798..1817)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1799..1817)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1904..1921)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2075..2663)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2155..2174)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2834..3625,3626..3694))
                     /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     3457..3476
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3695..3799)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3867..3885
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3923..3945)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4045..4064
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4145..4163
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4145..4162
                     /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 tgtttagctt
      421 gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac
      481 agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata
      541 catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa
      601 aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg
      661 aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg
      721 ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc
      781 gaacataaac aaccatggca gaaccagccc aaaaaaagca aaaacaaact gttcaggagc
      841 gcaaggcgtt tatctcccgt atcactaatg aaactaaaat tcaaatcgct atttcgctga
      901 atggtggtta tattcaaata aaagattcga ttcttcctgc aaagaaggat gacgatgtag
      961 cttcccaagc tactcagtca caggtcatcg atattcacac aggtgttggc tttttggatc
     1021 atatgatcca tgcgttggca aaacactctg gttggtctct tattgttgaa tgtatcggtg
     1081 acctgcacat tgacgatcac catactaccg aagattgcgg tatcgcatta gggcaagcgt
     1141 tcaaagaagc aatgggtgct gtccgtggtg taaaaagatt cggtactggg ttcgcaccat
     1201 tggatgaggc gctatcacgt gccgtagtcg atttatctaa tagaccattt gctgtaatcg
     1261 accttggatt gaagagagag atgattggtg atttatccac tgaaatgatt ccacactttt
     1321 tggaaagttt cgcggaggcg gccagaatta ctttgcatgt tgattgtctg agaggtttca
     1381 acgatcacca cagaagtgag agtgcgttca aggctttggc tgttgccata agagaagcta
     1441 tttctagcaa tggcaccaat gacgttccct caaccaaagg tgttttgatg tgaagtactg
     1501 acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt tatattgtag
     1561 ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc gacatcatct
     1621 gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc
     1681 tggtcgctat actgctgtcg attcgatact aacgccgcca tccagtttaa acgagctcga
     1741 attcatcgat gatatcagat ccactagtgg cctatgcggc cgcggatctg ccggtctccc
     1801 tatagtgagt cgtattaatt tcgataagcc aggttaacct gcattaatga atcggccaac
     1861 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     1921 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     1981 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2041 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2101 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2161 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2221 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     2281 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2341 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2401 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2461 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     2521 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2581 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2641 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2701 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     2761 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     2821 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     2881 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     2941 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3001 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3061 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3121 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3181 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3241 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3301 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3361 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3421 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     3481 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     3541 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     3601 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     3661 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     3721 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     3781 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     3841 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     3901 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     3961 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     4021 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     4081 tggacatatt gtcgttagaa cgcggctaca attaatacat aaccttatgt atcatacaca
     4141 tacgatttag gtgacactat aga
//
