LOCUS       sequence-9008-ex        4969 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 4969)
  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 4969)
  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-bleMX6-P81nmt1-3FLAG
FEATURES             Location/Qualifiers
     source          1..4969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..19)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2..19)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(100..119)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     219..241
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(279..297)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        365..469
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(470..538,539..1330)
                     /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     complement(688..707)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1501..2089
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1990..2009
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2243..2260
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2347..2366
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2347..2365
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            complement(2470..3391)
                     /label=bleMX6
                     /note="yeast selectable marker conferring phleomycin 
                     resistance"
     terminator      complement(2470..2667)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(2673..3047)
                     /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"
     promoter        complement(3048..3391)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        3436..4594
                     /label=nmt1 P81 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring weak thiamine-repressible expression"
     CDS             4612..4683
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     terminator      4707..4894
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4762..4786)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
ORIGIN
        1 tctatagtgt cacctaaatc gtatgtgtat gatacataag gttatgtatt aattgtagcc
       61 gcgttctaac gacaatatgt ccatatggtg cactctcagt acaatctgct ctgatgccgc
      121 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
      181 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
      241 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
      301 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
      361 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      421 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
      481 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
      541 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
      601 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
      661 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
      721 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
      781 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
      841 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
      901 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
      961 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     1021 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     1081 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     1141 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     1201 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     1261 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     1321 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     1381 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     1441 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1501 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1561 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1621 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     1681 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1741 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1801 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     1861 ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg
     1921 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     1981 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2041 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2101 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2161 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     2221 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     2281 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaggttaa cctggcttat
     2341 cgaaattaat acgactcact atagggagac cggcagatcc gcggccgcat aggccactag
     2401 tggatctgat atcatcgatg aattcgagct cgtttaaact ggatggcggc gttagtatcg
     2461 aatcgacagc agtatagcga ccagcattca catacgattg acgcatgata ttactttctg
     2521 cgcacttaac ttcgcatctg ggcagatgat gtcgaggcga aaaaaaatat aaatcacgct
     2581 aacatttgat taaaatagaa caactacaat ataaaaaaac tatacaaatg acaagttctt
     2641 gaaaacaaga atctttttat tgtcagtact gatcagtcct gctcctcggc cacgaagtgc
     2701 acgcagttgc cggccgggtc gcgcagggcg aactcccgcc cccacggctg ctcgccgatc
     2761 tcggtcatgg ccggcccgga ggcgtcccgg aagttcgtgg acacgacctc cgaccactcg
     2821 gcgtacagct cgtccaggcc gcgcacccac acccaggcca gggtgttgtc cggcaccacc
     2881 tggtcctgga ccgcgctgat gaacagggtc acgtcgtccc ggaccacacc ggcgaagtcg
     2941 tcctccacga agtcccggga gaacccgagc cggtcggtcc agaactcgac cgctccggcg
     3001 acgtcgcgcg cggtgagcac cggaacggca ctggtcaact tggccatggt tgtttatgtt
     3061 cggatgtgat gtgagaactg tatcctagca agattttaaa aggaagtata tgaaagaaga
     3121 acctcagtgg caaatcctaa ccttttatat ttctctacag gggcgcggcg tggggacaat
     3181 tcaacgcgtc tgtgagggga gcgtttccct gctcgcaggt ctgcagcgag gagccgtaat
     3241 ttttgcttcg cgccgtgcgg ccatcaaaat gtatggatgc aaatgattat acatggggat
     3301 gtatgggcta aatgtacggg cgacagtcac atcatgcccc tgagctgcgc acgtcaagac
     3361 tgtcaaggag ggtattctgg gcctccatgt cgctggccgg gtgacccggc ggggacgagg
     3421 caagctaaac agatctcgcc ataaaagaca gaataagtca tcagcggttg tttcatttcc
     3481 tatatttttt ttttattttt ttatttttta ataagggaaa atttaacgtc taaggataca
     3541 gaagattgtt agcacattaa agtaataaag gcttaagtag taagtgcctt agcatgttat
     3601 tgtatttcaa aggacataat ctaaaataat aacaatatca tttctcacaa gttattcaat
     3661 tttctttttt ttttctaata atatcaagaa tgtattattt gtttgacata agtcaactaa
     3721 tttatttaat atgctggatt aatcttgcag acatgtaaat taacaagttt tagtcaaata
     3781 acgttgaagt ttcaatgaac tcaaataatt tctctttttt tttatataac catatatgtc
     3841 taatctgatt tatattttcc gcaggatcaa ctgaagttat gacatttgga ttggatcact
     3901 tataaccttg gtcgccaaat aatacaaaaa tcagcgttat aaaacaaaga aggtttttgt
     3961 taagaaatta atcctctttc ttgataagaa agttgaaccg aaattgcaga tactgatata
     4021 tgaaaataat acccacaatt ttgggaatag cgcaagcctc aatttaaaca ataggtgagg
     4081 acacatgata atgacctcaa tgattgttag aagaaaagag cctcattaca aaatcgaaaa
     4141 atgaatggtt gggtacaagt ttccaaaaca tggtaaagtg gactttgcgt atgagacgta
     4201 aatagaaaaa aacacttgtt atatgttttc tagaattatt gttgtctctt tatggttgga
     4261 tgatgcaaaa tagtaatttc ggttagttgc tgtaaaacac cacgagacaa atagatatgg
     4321 atatttatta aatcaggaaa aacgtaactc tcggctactg gatggttcag tcacccaacg
     4381 attactgggg agagaaaaca gggcaaaagc aaagcttaaa ggaatccgat tgtcattcgg
     4441 caatgtgcag cgaaactaaa aaccggataa tggacctgtt aatcgaaaca ttgaagatgg
     4501 aagaggaatc ctggcatatc atcaattgaa taagttgaat taattatttc aatctcattc
     4561 tcactttctg acttatagtc gctttgttaa atcatgtctt taattaatat ggactacaag
     4621 gacgacgatg acaaggatta caaagatgac gacgataagg actataagga cgatgatgat
     4681 aaatagggcg cgccacttct aaataagcga atttcttatg atttatgatt tttattatta
     4741 aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac
     4801 gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagt
     4861 atgaggtcgc tcttattgac cacacctcta ccggcagatc cgctagggat aacagggtaa
     4921 tatagatccg tcgacctgca gcgtacgaag cttcagctgg cggccgcgt
//
