LOCUS       sequence-466681-        4969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for generating integration cassette to C-terminally tag gene
            of interest with FAPoptim::KANMX.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4969)
  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 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: "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-KAN
FEATURES             Location/Qualifiers
     source          1..4969
                     /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..2326)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      complement(970..1167)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(1173..1982)
                     /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     1896..1915
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1983..2326)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      complement(2401..2588)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     2509..2533
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2612..2641)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(2642..2671)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(3470..3488)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3471..3488)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3569..3588)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3688..3710
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3748..3766)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3834..3938
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3939..4007,4008..4799)
                     /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(4157..4176)
                     /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 gattagaaaa actcatcgag catcaaatga
     1201 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
     1261 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
     1321 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
     1381 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagctta
     1441 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
     1501 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg
     1561 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
     1621 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttg
     1681 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     1741 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
     1801 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     1861 aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     1921 aaatcagcat ccatgttgga atttaatcgc ggcctcgaaa cgtgagtctt ttccttaccc
     1981 atggttgttt atgttcggat gtgatgtgag aactgtatcc tagcaagatt ttaaaaggaa
     2041 gtatatgaaa gaagaacctc agtggcaaat cctaaccttt tatatttctc tacaggggcg
     2101 cggcgtgggg acaattcaac gcgtctgtga ggggagcgtt tccctgctcg caggtctgca
     2161 gcgaggagcc gtaatttttg cttcgcgccg tgcggccatc aaaatgtatg gatgcaaatg
     2221 attatacatg gggatgtatg ggctaaatgt acgggcgaca gtcacatcat gcccctgagc
     2281 tgcgcacgtc aagactgtca aggagggtat tctgggcctc catgtcgctg gccgggtgac
     2341 ccggcgggga cgaggcaagc taaacagatc tatattaccc tgttatccct agcggatctg
     2401 ccggtagagg tgtggtcaat aagagcgacc tcatactata cctgagaaag caacctgacc
     2461 tacaggaaag agttactcaa gaataagaat tttcgtttta aaacctaaga gtcactttaa
     2521 aatttgtata cacttatttt ttttataact tatttaataa taaaaatcat aaatcataag
     2581 aaattcgctt atttagaagt ggcgcgcctt acaggtcctc ctcgctgatc agcttctgct
     2641 ccaggtcctc ctcgctgatc agcttctgct cagacaaaac agtcaattga gtaccaccac
     2701 cgaacaagta accgtcaacg tcagacaaag aacagtagta ttcagcttcg tcttctggtt
     2761 gagcgtcaga gatagtcaaa gcagccttag cacccaacaa agaaccagag aatctacctg
     2821 gagtccaaga gtacttcttg tcagtgtcga agatcaaagc tcttggagct tgacctggct
     2881 tttgttggaa ccagttagcg tagtgaccag aagtaacagc accagtacca gaaccacaag
     2941 tcaagataac agtaccacct ggagaaacag taacagatgg ttcttgagta acaacagctt
     3001 gagaaccacc accaccagaa ccaccaccac cagaaccacc accaccagaa ccaccaccac
     3061 cagacaaaac agtcaattga gtaccaccac cgaacaagta accgtcaacg tcagacaaag
     3121 aacagtagta ttcagcttcg tcttctggtt gagcgtcaga gatagtcaaa gcagccttag
     3181 cacccaacaa agaaccagag aatctacctg gagtccaaga gtacttcttg tcagtgtcga
     3241 agatcaaagc tcttggagct tgacctggct tttgttggaa ccagttagcg tagtgaccag
     3301 aagtaacagc accagtacca gaaccacaag tcaagataac agtaccacct ggagaaacag
     3361 taacagatgg ttcttgagta acaacagctt gacccgcacc cgcacccgcg ttaattaacc
     3421 cggggatccg tcgacctgca gcgtacgaag cttcagctgg cggccgcgtt ctatagtgtc
     3481 acctaaatcg tatgtgtatg atacataagg ttatgtatta attgtagccg cgttctaacg
     3541 acaatatgtc catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3601 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3661 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3721 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     3781 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3841 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3901 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3961 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4021 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4081 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4141 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4201 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4261 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4321 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4381 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4441 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4501 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4561 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4621 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4681 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4741 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4801 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4861 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4921 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttc
//
