LOCUS       sequence-142646-        4905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  alias pFA6a-mCherry-hphNT1 - vector for C-terminal tagging of yeast 
            genes with mCherry and hygromycin selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4905)
  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 4905)
  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: pFM689
FEATURES             Location/Qualifiers
     source          1..4905
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        72..415
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             416..1444
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             416..1444
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      1452..1699
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1468..1486)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1761..1780)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1762..1780)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1867..1884)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2038..2626)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2118..2137)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2797..3588,3589..3657))
                     /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     3420..3439
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3658..3762)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3830..3848
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3886..3908)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4008..4027
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4108..4126
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4108..4125
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4194..4904
                     /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(4339..4357)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4607..4626
                     /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 ggacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc cacgccgcgc
      301 ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt catatacttc
      361 cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa caaccatggg
      421 taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
      481 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
      541 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
      601 ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg
      661 ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
      721 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc
      781 gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
      841 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
      901 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
      961 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     1021 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     1081 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     1141 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg
     1201 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     1261 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     1321 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg
     1381 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     1441 ataatctcga gtcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc
     1501 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     1561 tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta
     1621 cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg
     1681 ctcgaaggct ttaacgagct cgaattcatc gatgatatca gatccactag tggcctatgc
     1741 ggccgcggat ctgccggtct ccctatagtg agtcgtatta atttcgataa gccaggttaa
     1801 cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     1861 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1921 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1981 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2041 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2101 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2161 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2221 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2281 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2341 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2401 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2461 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2521 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2581 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2641 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2701 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2761 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2821 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2881 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2941 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3001 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3061 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3121 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3181 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3241 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3301 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3361 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3421 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3481 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3541 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3601 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3661 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3721 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3781 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     3841 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     3901 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     3961 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     4021 gattgtactg agagtgcacc atatggacat attgtcgtta gaacgcggct acaattaata
     4081 cataacctta tgtatcatac acatacgatt taggtgacac tatagaacgc ggccgccagc
     4141 tgaagcttcg tacgctgcag gtcgacggtg caggtgctgg agctggtgct ggtatggtga
     4201 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     4261 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     4321 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     4381 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     4441 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     4501 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     4561 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     4621 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     4681 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     4741 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     4801 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     4861 agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaag
//
