LOCUS       sequence-148903-        4519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  alias pFA6a-mCherry-natNT2 - vector for C-terminal tagging of yeast 
            genes with mCherry and nourseothricin selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4519)
  AUTHORS   Malcova I, Farkasovsky M, Senohrabkova L, Vasicova P, Hasek J
  TITLE     New integrative modules for multicolor-protein labeling and 
            live-cell imaging in Saccharomyces cerevisiae.
  JOURNAL   FEMS Yeast Res. 2016 Mar 17. pii: fow027.
  PUBMED    26994102
REFERENCE   2  (bases 1 to 4519)
  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: "FEMS Yeast 
            Res. 2016 Mar 17. pii: fow027."
COMMENT     Sequence Label: pFM699
FEATURES             Location/Qualifiers
     source          1..4519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        72..415
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             416..988
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      996..1183
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1051..1075)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1375..1394)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1376..1394)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1481..1498)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1652..2240)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1732..1751)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2411..3202,3203..3271))
                     /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     3034..3053
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3272..3376)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3444..3462
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3500..3522)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3622..3641
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3722..3740
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3722..3739
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             3808..4518
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3953..3971)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4221..4240
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 gatccccggg ttaattaagg cgcgccagat ctgtttagct tgcctcgtcc ccgccgggtc
       61 acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac gtgcgcagct
      121 caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat gtataatcat
      181 ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc
      241 agacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc cacgccgcgc
      301 ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt catatacttc
      361 cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa caaccatggg
      421 taccactctt gacgacacgg cttaccggta ccgcaccagt gtcccggggg acgccgaggc
      481 catcgaggca ctggatgggt ccttcaccac cgacaccgtc ttccgcgtca ccgccaccgg
      541 ggacggcttc accctgcggg aggtgccggt ggacccgccc ctgaccaagg tgttccccga
      601 cgacgaatcg gacgacgaat cggacgacgg ggaggacggc gacccggact cccggacgtt
      661 cgtcgcgtac ggggacgacg gcgacctggc gggcttcgtg gtcgtctcgt actccggctg
      721 gaaccgccgg ctgaccgtcg aggacatcga ggtcgccccg gagcaccggg ggcacggggt
      781 cgggcgcgcg ttgatggggc tcgcgacgga gttcgcccgc gagcggggcg ccgggcacct
      841 ctggctggag gtcaccaacg tcaacgcacc ggcgatccac gcgtaccggc ggatggggtt
      901 caccctctgc ggcctggaca ccgccctgta cgacggcacc gcctcggacg gcgagcaggc
      961 gctctacatg agcatgccct gcccctaatc tcgaggcgaa tttcttatga tttatgattt
     1021 ttattattaa ataagttata aaaaaaataa gtgtatacaa attttaaagt gactcttagg
     1081 ttttaaaacg aaaattctta ttcttgagta actctttcct gtaggtcagg ttgctttctc
     1141 aggtatagca tgaggtcgct cttattgacc acacctctac cggcatgccg agcaaatgcc
     1201 tgcaaatcgc tccccatttc acccaattgt agatatgcta actccagcaa tgagttgatg
     1261 aatctcggtg tgtattttat gtcctcagag gacaacacct gttgtaatcg agctcgaatt
     1321 catcgatgat atcagatcca ctagtggcct atgcggccgc ggatctgccg gtctccctat
     1381 agtgagtcgt attaatttcg ataagccagg ttaacctgca ttaatgaatc ggccaacgcg
     1441 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     1501 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1561 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     1621 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1681 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1741 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1801 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     1861 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1921 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1981 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2041 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     2101 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2161 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2221 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2281 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2341 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2401 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2461 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     2521 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     2581 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     2641 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     2701 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     2761 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     2821 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     2881 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     2941 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3001 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3061 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3121 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3181 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3241 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     3301 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     3361 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     3421 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     3481 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     3541 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     3601 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgg
     3661 acatattgtc gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac
     3721 gatttaggtg acactataga acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac
     3781 ggtgcaggtg ctggagctgg tgctggtatg gtgagcaagg gcgaggagga taacatggcc
     3841 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     3901 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     3961 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     4021 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     4081 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     4141 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     4201 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     4261 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     4321 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     4381 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     4441 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     4501 gacgagctgt acaagtaag
//
