LOCUS       sequence-466721-        4732 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for generating integration cassette to C-terminally tag gene
            of interest with FAPoptim::NATMX.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4732)
  AUTHORS   Oppenheimer KG, Hager NA, McAtee CK, Filiztekin E, Shang C, Warnick 
            JA, Bruchez MP, Brodsky JL, Prosser DC, Kwiatkowski AV, O'Donnell AF
  TITLE     Optimization of the fluorogen-activating protein tag for 
            quantitative protein trafficking and colocalization studies in S. 
            cerevisiae.
  JOURNAL   Mol Biol Cell. 2024 Jul 1;35(7):mr5. doi: 10.1091/mbc.E24-04-0174. 
            Epub 2024 May 29.
  PUBMED    38809589
REFERENCE   2  (bases 1 to 4732)
  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: "Mol Biol 
            Cell."; date: "2024-07-1"; volume: "35(7):mr5. doi"; pages: " 
            10.1091/mbc.E24-04-0174. Epub 2024 May 29"
COMMENT     Sequence Label: pFA6-CFAPoptim-NAT
FEATURES             Location/Qualifiers
     source          1..4732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     847..866
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        847..865
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            complement(970..2089)
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     terminator      complement(970..1167)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(1173..1745)
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     promoter        complement(1746..2089)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      complement(2164..2351)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     2272..2296
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2375..2404)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(2405..2434)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(3233..3251)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3234..3251)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3332..3351)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3451..3473
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3511..3529)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3597..3701
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3702..3770,3771..4562)
                     /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(3920..3939)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaggttaa cctggcttat
      841 cgaaattaat acgactcact atagggagac cggcagatcc gcggccgcat aggccactag
      901 tggatctgat atcatcgatg aattcgagct cgtttaaact ggatggcggc gttagtatcg
      961 aatcgacagc agtatagcga ccagcattca catacgattg acgcatgata ttactttctg
     1021 cgcacttaac ttcgcatctg ggcagatgat gtcgaggcga aaaaaaatat aaatcacgct
     1081 aacatttgat taaaatagaa caactacaat ataaaaaaac tatacaaatg acaagttctt
     1141 gaaaacaaga atctttttat tgtcagtact gattaggggc agggcatgct catgtagagc
     1201 gcctgctcgc cgtccgaggc ggtgccgtcg tacagggcgg tgtccaggcc gcagagggtg
     1261 aaccccatcc gccggtacgc gtggatcgcc ggtgcgttga cgttggtgac ctccagccag
     1321 aggtgcccgg cgccccgctc gcgggcgaac tccgtcgcga gccccatcaa cgcgcgcccg
     1381 accccgtgcc cccggtgctc cggggcgacc tcgatgtcct cgacggtcag ccggcggttc
     1441 cagccggagt acgagacgac cacgaagccc gccaggtcgc cgtcgtcccc gtacgcgacg
     1501 aacgtccggg agtccgggtc gccgtcctcc ccgtcgtccg attcgtcgtc cgattcgtcg
     1561 tcggggaaca ccttggtcag gggcgggtcc accggcacct cccgcagggt gaagccgtcc
     1621 ccggtggcgg tgacgcggaa gacggtgtcg gtggtgaagg acccatccag tgcctcgatg
     1681 gcctcggcgt cccccgggac actggtgcgg taccggtaag ccgtatcgtc aagagtggta
     1741 cccatggttg tttatgttcg gatgtgatgt gagaactgta tcctagcaag attttaaaag
     1801 gaagtatatg aaagaagaac ctcagtggca aatcctaacc ttttatattt ctctacaggg
     1861 gcgcggcgtg gggacaattc aacgcgtctg tgaggggagc gtttccctgc tcgcaggtct
     1921 gcagcgagga gccgtaattt ttgcttcgcg ccgtgcggcc atcaaaatgt atggatgcaa
     1981 atgattatac atggggatgt atgggctaaa tgtacgggcg acagtcacat catgcccctg
     2041 agctgcgcac gtcaagactg tcaaggaggg tattctgggc ctccatgtcg ctggccgggt
     2101 gacccggcgg ggacgaggca agctaaacag atctatatta ccctgttatc cctagcggat
     2161 ctgccggtag aggtgtggtc aataagagcg acctcatact atacctgaga aagcaacctg
     2221 acctacagga aagagttact caagaataag aattttcgtt ttaaaaccta agagtcactt
     2281 taaaatttgt atacacttat tttttttata acttatttaa taataaaaat cataaatcat
     2341 aagaaattcg cttatttaga agtggcgcgc cttacaggtc ctcctcgctg atcagcttct
     2401 gctccaggtc ctcctcgctg atcagcttct gctcagacaa aacagtcaat tgagtaccac
     2461 caccgaacaa gtaaccgtca acgtcagaca aagaacagta gtattcagct tcgtcttctg
     2521 gttgagcgtc agagatagtc aaagcagcct tagcacccaa caaagaacca gagaatctac
     2581 ctggagtcca agagtacttc ttgtcagtgt cgaagatcaa agctcttgga gcttgacctg
     2641 gcttttgttg gaaccagtta gcgtagtgac cagaagtaac agcaccagta ccagaaccac
     2701 aagtcaagat aacagtacca cctggagaaa cagtaacaga tggttcttga gtaacaacag
     2761 cttgagaacc accaccacca gaaccaccac caccagaacc accaccacca gaaccaccac
     2821 caccagacaa aacagtcaat tgagtaccac caccgaacaa gtaaccgtca acgtcagaca
     2881 aagaacagta gtattcagct tcgtcttctg gttgagcgtc agagatagtc aaagcagcct
     2941 tagcacccaa caaagaacca gagaatctac ctggagtcca agagtacttc ttgtcagtgt
     3001 cgaagatcaa agctcttgga gcttgacctg gcttttgttg gaaccagtta gcgtagtgac
     3061 cagaagtaac agcaccagta ccagaaccac aagtcaagat aacagtacca cctggagaaa
     3121 cagtaacaga tggttcttga gtaacaacag cttgacccgc acccgcaccc gcgttaatta
     3181 acccggggat ccgtcgacct gcagcgtacg aagcttcagc tggcggccgc gttctatagt
     3241 gtcacctaaa tcgtatgtgt atgatacata aggttatgta ttaattgtag ccgcgttcta
     3301 acgacaatat gtccatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3361 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3421 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3481 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     3541 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     3601 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3661 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3721 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3781 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3841 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3901 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     3961 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4021 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4081 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4141 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4201 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4261 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4321 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4381 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4441 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4501 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4561 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4621 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4681 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tc
//
