LOCUS       sequence-8998-ex        4078 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 4078)
  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 4078)
  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-natMX6
FEATURES             Location/Qualifiers
     source          1..4078
                     /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..1611
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        489..832
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             833..1408
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVISYSAWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFAGERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGERQALYMSMPCP"
     terminator      1414..1611
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1715..1734)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1716..1734)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1821..1838)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1992..2580)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2072..2091)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2751..3542,3543..3611))
                     /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     3374..3393
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3612..3716)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3784..3802
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3840..3862)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3962..3981
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4062..4078,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4062..4078,1)
                     /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 ccatgggtac
      841 cactcttgac gacacggctt accggtaccg caccagtgtc ccgggggacg ccgaggccat
      901 cgaggcactg gatgggtcct tcaccaccga caccgtcttc cgcgtcaccg ccaccgggga
      961 cggcttcacc ctgcgggagg tgccggtgga cccgcccctg accaaggtgt tccccgacga
     1021 cgaatcggac gacgaatcgg acgacgggga ggacggcgac ccggactccc ggacgttcgt
     1081 cgcgtacggg gacgacggcg acctggcggg cttcgtggtc atctcgtact cggcgtggaa
     1141 ccgccggctg accgtcgagg acatcgaggt cgccccggag caccgggggc acggggtcgg
     1201 gcgcgcgttg atggggctcg cgacggagtt cgccggcgag cggggcgccg ggcacctctg
     1261 gctggaggtc accaacgtca acgcaccggc gatccacgcg taccggcgga tggggttcac
     1321 cctctgcggc ctggacaccg ccctgtacga cggcaccgcc tcggacggcg agcggcaggc
     1381 gctctacatg agcatgccct gcccctaatc agtactgaca ataaaaagat tcttgttttc
     1441 aagaacttgt catttgtata gtttttttat attgtagttg ttctatttta atcaaatgtt
     1501 agcgtgattt atattttttt tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg
     1561 cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg tcgctatact gctgtcgatt
     1621 cgatactaac gccgccatcc agtttaaacg agctcgaatt catcgatgat atcagatcca
     1681 ctagtggcct atgcggccgc ggatctgccg gtctccctat agtgagtcgt attaatttcg
     1741 ataagccagg ttaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     1801 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     1861 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     1921 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1981 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2041 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2101 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2161 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     2221 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2281 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2341 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2401 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     2461 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2521 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2581 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2641 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     2701 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     2761 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     2821 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     2881 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     2941 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3001 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3061 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3121 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3181 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3241 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3301 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     3361 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     3421 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     3481 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     3541 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     3601 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     3661 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     3721 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     3781 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     3841 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     3901 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     3961 gcggcatcag agcagattgt actgagagtg caccatatgg acatattgtc gttagaacgc
     4021 ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg acactata
//
