LOCUS       sequence-256799-        4312 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 4312)
  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 4312)
  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-KanMX6
FEATURES             Location/Qualifiers
     source          1..4312
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             86..217
                     /codon_start=1
                     /product="five tandem FLAG(R) epitope tags"
                     /label=5xFLAG
                     /translation="DYKDDDDKDYKDDDDKLMDYKDDDDKDYKDDDDKLMDYKDDDDK"
     terminator      244..431
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(299..323)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            506..1862
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        506..849
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             850..1659
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(917..936)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1665..1862
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1966..1985)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1967..1985)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2072..2089)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2243..2831)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2323..2342)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3002..3793,3794..3862))
                     /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     3625..3644
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3863..3967)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4035..4053
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4091..4113)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4213..4232
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atttaggtga cactatagaa cgcggccgcc agctgaagct tcgtacgctg caggtcgacg
       61 gatccccggg ttaattaatc atatggacta caaggacgac gatgacaagg attacaaaga
      121 tgacgacgat aagcttatgg actacaagga cgacgatgac aaggattaca aagatgacga
      181 cgataagctt atggactaca aggacgacga tgacaagcat atgggcgcgc cacttctaaa
      241 taagcgaatt tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt
      301 gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac
      361 tctttcctgt aggtcaggtt gctttctcag gtatagtatg aggtcgctct tattgaccac
      421 acctctaccg gcagatccgc tagggataac agggtaatat agatctgttt agcttgcctc
      481 gtccccgccg ggtcacccgg ccagcgacat ggaggcccag aataccctcc ttgacagtct
      541 tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg tacatttagc ccatacatcc
      601 ccatgtataa tcatttgcat ccatacattt tgatggccgc acggcgcgaa gcaaaaatta
      661 cggctcctcg ctgcagacct gcgagcaggg aaacgctccc ctcacagacg cgttgaattg
      721 tccccacgcc gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt
      781 ctttcatata cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca
      841 taaacaacca tgggtaagga aaagactcac gtttcgaggc cgcgattaaa ttccaacatg
      901 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
      961 atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     1021 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     1081 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     1141 gcgatccccg gcaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     1201 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     1261 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     1321 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     1381 aaagaaatgc ataagctttt gccattctca ccggattcag tcgtcactca tggtgatttc
     1441 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     1501 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     1561 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     1621 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagtactgac aataaaaaga
     1681 ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt
     1741 aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga
     1801 agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac
     1861 tgctgtcgat tcgatactaa cgccgccatc cagtttaaac gagctcgaat tcatcgatga
     1921 tatcagatcc actagtggcc tatgcggccg cggatctgcc ggtctcccta tagtgagtcg
     1981 tattaatttc gataagccag gttaacctgc attaatgaat cggccaacgc gcggggagag
     2041 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2101 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2161 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2221 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2281 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2341 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2401 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2461 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2521 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2581 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2641 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2701 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2761 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2821 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2881 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2941 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3001 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3061 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3121 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3181 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3241 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3301 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3361 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3421 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3481 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3541 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3601 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3661 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3721 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3781 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3841 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     3901 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3961 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     4021 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     4081 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     4141 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     4201 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg gacatattgt
     4261 cgttagaacg cggctacaat taatacataa ccttatgtat catacacata cg
//
