LOCUS       sequence-61667-e        4177 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 4177)
  TITLE     Formosa lab yeast 3' tagging constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4177)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pTF264 (pFA6a-TEV-6xGly-3xFlag-His3MX)
FEATURES             Location/Qualifiers
     source          1..4177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(199..303)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     371..389
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(427..449)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     549..568
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        649..667
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     649..666
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             728..748
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             776..841
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      870..1057
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(925..949)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1132..1475
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1476..2174
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSNRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      2178..2375
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2479..2498)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2480..2498)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2585..2602)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2756..3344)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2836..2855)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3515..4177,1..129,130..198))
                     /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     4138..4157
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
       61 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
      121 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
      181 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
      241 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
      301 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
      361 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
      421 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
      481 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
      541 taactatgcg gcatcagagc agattgtact gagagtgcac catatggaca tattgtcgtt
      601 agaacgcggc tacaattaat acataacctt atgtatcata cacatacgat ttaggtgaca
      661 ctatagaacg cggccgccag ctgaagcttc gtacgctgca ggtcgacgga tccccgggtt
      721 aattaacgag aatttatact tccaaggttc tggtggaggc ggtggaggct ctagagacta
      781 caaggaccac gacggtgact acaaggacca cgacattgac tacaaggatg acgatgacaa
      841 gtgactcgag gcgcgccact tctaaataag cgaatttctt atgatttatg atttttatta
      901 ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa
      961 aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat
     1021 agtatgaggt cgctcttatt gaccacacct ctaccggcag atccgctagg gataacaggg
     1081 taatatagat ctgtttagct tgcctcgtcc ccgccgggtc acccggccag cgacatggag
     1141 gcccagaata ccctccttga cagtcttgac gtgcgcagct caggggcatg atgtgactgt
     1201 cgcccgtaca tttagcccat acatccccat gtataatcat ttgcatccat acattttgat
     1261 ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc agacctgcga gcagggaaac
     1321 gctcccctca cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag aaatataaaa
     1381 ggttaggatt tgccactgag gttcttcttt catatacttc cttttaaaat cttgctagga
     1441 tacagttctc acatcacatc cgaacataaa caaccatggc agaaccagcc caaaaaaagc
     1501 aaaaacaaac tgttcaggag cgcaaggcgt ttatctcccg tatcactaat gaaactaaaa
     1561 ttcaaatcgc tatttcgctg aatggtggtt atattcaaat aaaagattcg attcttcctg
     1621 caaagaagga tgacgatgta gcttcccaag ctactcagtc acaggtcatc gatattcaca
     1681 caggtgttgg ctttttggat catatgatcc atgcgttggc aaaacactct ggttggtctc
     1741 ttattgttga atgtatcggt gacctgcaca ttgacgatca ccatactacc gaagattgcg
     1801 gtatcgcatt agggcaagcg ttcaaagaag caatgggtgc tgtccgtggt gtaaaaagat
     1861 tcggtactgg gttcgcacca ttggatgagg cgctatcacg tgccgtagtc gatttatcta
     1921 atagaccatt tgctgtaatc gaccttggat tgaagagaga gatgattggt gatttatcca
     1981 ctgaaatgat tccacacttt ttggaaagtt tcgcggaggc ggccagaatt actttgcatg
     2041 ttgattgtct gagaggtttc aacgatcacc acagaagtga gagtgcgttc aaggctttgg
     2101 ctgttgccat aagagaagct atttctagca atggcaccaa tgacgttccc tcaaccaaag
     2161 gtgttttgat gtgaagtact gacaataaaa agattcttgt tttcaagaac ttgtcatttg
     2221 tatagttttt ttatattgta gttgttctat tttaatcaaa tgttagcgtg atttatattt
     2281 tttttcgcct cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat
     2341 gcgtcaatcg tatgtgaatg ctggtcgcta tactgctgtc gattcgatac taacgccgcc
     2401 atccagttta aacgagctcg aattcatcga tgatatcaga tccactagtg gcctatgcgg
     2461 ccgcggatct gccggtctcc ctatagtgag tcgtattaat ttcgataagc caggttaacc
     2521 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2581 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2641 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2701 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2761 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2821 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2881 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2941 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3001 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3061 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3121 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3181 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3241 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3301 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3361 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3421 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3481 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3541 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3601 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3661 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3721 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3781 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3841 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3901 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3961 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4021 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4081 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4141 ataccgcgcc acatagcaga actttaaaag tgctcat
//
