LOCUS       sequence-8999-ex        3877 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 3877)
  AUTHORS   Noguchi C, Garabedian MV, Malik M, Noguchi E
  TITLE     A vector system for genomic FLAG epitope-tagging in 
            Schizosaccharomyces pombe.
  JOURNAL   Biotechnol J. 2008 Oct;3(9-10):1280-5. doi: 10.1002/biot.200800140.
  PUBMED    18729046
REFERENCE   2  (bases 1 to 3877)
  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: "Biotechnol 
            J."; date: "2008-10"; volume: "3(9-10):1280-5. doi"; pages: " 
            10.1002/biot.200800140"
COMMENT     Sequence Label: pFA6a-5FLAG-bleMX6
FEATURES             Location/Qualifiers
     source          1..3877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             69..200
                     /codon_start=1
                     /product="five tandem FLAG(R) epitope tags"
                     /label=5xFLAG
                     /translation="DYKDDDDKDYKDDDDKLMDYKDDDDKDYKDDDDKLMDYKDDDDK"
     terminator      227..414
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(282..306)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            489..1410
                     /label=bleMX6
                     /note="yeast selectable marker conferring phleomycin 
                     resistance"
     promoter        489..832
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             833..1207
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      1213..1410
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1514..1533)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1515..1533)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1620..1637)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1791..2379)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1871..1890)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2550..3341,3342..3410))
                     /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     3173..3192
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3411..3515)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3583..3601
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3639..3661)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3761..3780
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(3861..3877,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(3861..3877,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 atcatatgga ctacaaggac gacgatgaca aggattacaa agatgacgac gataagctta
      121 tggactacaa ggacgacgat gacaaggatt acaaagatga cgacgataag cttatggact
      181 acaaggacga cgatgacaag catatgggcg cgccacttct aaataagcga atttcttatg
      241 atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag
      301 tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc tgtaggtcag
      361 gttgctttct caggtatagt atgaggtcgc tcttattgac cacacctcta ccggcagatc
      421 cgctagggat aacagggtaa tatagatctg tttagcttgc ctcgtccccg ccgggtcacc
      481 cggccagcga catggaggcc cagaataccc tccttgacag tcttgacgtg cgcagctcag
      541 gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta taatcatttg
      601 catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga
      661 cctgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac gccgcgcccc
      721 tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat atacttcctt
      781 ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa ccatggccaa
      841 gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg
      901 gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg
      961 ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct
     1021 ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc
     1081 cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg
     1141 gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca
     1201 ggactgatca gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag
     1261 tttttttata ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt
     1321 cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc
     1381 aatcgtatgt gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatcca
     1441 gtttaaacga gctcgaattc atcgatgata tcagatccac tagtggccta tgcggccgcg
     1501 gatctgccgg tctccctata gtgagtcgta ttaatttcga taagccaggt taacctgcat
     1561 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     1621 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1681 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1741 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1801 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1861 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1921 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1981 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2041 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2101 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2161 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2221 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2281 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2341 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2401 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2461 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2521 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2581 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2641 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     2701 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2761 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     2821 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     2881 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     2941 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3001 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3061 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3121 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3181 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3241 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3301 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3361 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3421 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3481 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3541 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     3601 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     3661 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     3721 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     3781 ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta atacataacc
     3841 ttatgtatca tacacatacg atttaggtga cactata
//
