LOCUS       sequence-252098-        4731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For tagging genes at 3'end (C-terminal end of protein) in S. 
            cerevisiae with mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4731)
  TITLE     mNeonGreen tagging tools for yeast
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4731)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pFA6a-mNeonGreen-HIS3MX6
FEATURES             Location/Qualifiers
     source          1..4731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        77..95
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     77..94
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      895..1082
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(950..974)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1157..2357
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        1157..1500
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1501..2154
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHTNQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMSNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     terminator      2160..2357
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2461..2480)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2462..2480)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2567..2584)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2738..3326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2818..2837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3497..4288,4289..4357))
                     /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     4120..4139
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4358..4462)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4530..4548
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4586..4608)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4708..4727
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gagtgcacca tatggacata ttgtcgttag aacgcggcta caattaatac ataaccttat
       61 gtatcataca catacgattt aggtgacact atagaacgcg gccgccagct gaagcttcgt
      121 acgctgcagg tcgacggatc ggtgacggtg ctggtttaat taaatggtga gtaaaggcga
      181 ggaggacaat atggcgagct tacctgctac ccacgaactg cacattttcg gctcaatcaa
      241 tggtgtagat tttgacatgg tcggtcaggg gactggtaac cctaacgatg gttatgagga
      301 gctaaatctg aaatctacta aaggggacct tcaatttagt ccctggatac ttgttccgca
      361 cataggttat ggtttccacc aatatttgcc ctaccctgac gggatgagcc ccttccaggc
      421 ggctatggta gatggcagtg gctatcaggt ccacagaaca atgcaatttg aggacggggc
      481 cagcctaacc gtgaattata ggtacacgta cgaggggtca catataaagg gcgaagcaca
      541 ggtgaagggg acggggttcc cggcggacgg accagtgatg acaaactccc tgaccgctgc
      601 cgattggtgc cgtagtaaga agacttatcc taacgacaaa acgataatca gtaccttcaa
      661 gtggagttat accacgggca acgggaaaag ataccgtagc acggcgagaa caacttatac
      721 tttcgccaag ccaatggctg ccaactactt gaagaatcag cccatgtatg tattccgtaa
      781 gacggagctg aaacatagca aaaccgaact aaacttcaag gagtggcaga aagcgtttac
      841 cgatgtgatg ggtatggacg aattgtataa gtaaggcgcg ccacttctaa ataagcgaat
      901 ttcttatgat ttatgatttt tattattaaa taagttataa aaaaaataag tgtatacaaa
      961 ttttaaagtg actcttaggt tttaaaacga aaattcttat tcttgagtaa ctctttcctg
     1021 taggtcaggt tgctttctca ggtatagtat gaggtcgctc ttattgacca cacctctacc
     1081 ggcagatccg ctagggataa cagggtaata tagatctgtt tagcttgcct cgtccccgcc
     1141 gggtcacccg gccagcgaca tggaggccca gaataccctc cttgacagtc ttgacgtgcg
     1201 cagctcaggg gcatgatgtg actgtcgccc gtacatttag cccatacatc cccatgtata
     1261 atcatttgca tccatacatt ttgatggccg cacggcgcga agcaaaaatt acggctcctc
     1321 gctgcagacc tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt gtccccacgc
     1381 cgcgcccctg tagagaaata taaaaggtta ggatttgcca ctgaggttct tctttcatat
     1441 acttcctttt aaaatcttgc taggatacag ttctcacatc acatccgaac ataaacaacc
     1501 atgggtagga gggcttttgt agaaagaaat acgaacgaaa cgaaaatcag cgttgccatc
     1561 gctttggaca aagctccctt acctgaagag tcgaatttta ttgatgaact tataacttcc
     1621 aagcatacaa accaaaaggg agaacaagta atccaagtag acacgggaat tggattcttg
     1681 gatcacatgt atcatgcact ggctaaacat gcaggctgga gcttacgact ttactcaaga
     1741 ggtgatttaa tcatcgatga tcatcacact gcagaagata ctgctattgc acttggtatt
     1801 gcattcaagc aggctatgag taactttgcc ggcgttaaaa gatttggaca tgcttattgt
     1861 ccacttgacg aagctctttc tagaagcgta gttgacttgt cgggacggcc ctatgctgtt
     1921 atcgatttgg gattaaagcg tgaaaaggtt ggggaattgt cctgtgaaat gatccctcac
     1981 ttactatatt ccttttcggt agcagctgga attactttgc atgttacctg cttatatggt
     2041 agtaatgacc atcatcgtgc tgaaagcgct tttaaatctc tggctgttgc catgcgcgcg
     2101 gctactagtc ttactggaag ttctgaagtc ccaagcacga agggagtgtt gtaaagagta
     2161 ctgacaataa aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg
     2221 tagttgttct attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca
     2281 tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa
     2341 tgctggtcgc tatactgctg tcgattcgat actaacgccg ccatccagtt taaacgagct
     2401 cgaattcatc gatgatatca gatccactag tggcctatgc ggccgcggat ctgccggtct
     2461 ccctatagtg agtcgtatta atttcgataa gccaggttaa cctgcattaa tgaatcggcc
     2521 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2581 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2641 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2701 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2761 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2821 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2881 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2941 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3001 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3061 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3121 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3181 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3241 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3301 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3361 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3421 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3481 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3541 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3601 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3661 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3721 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3781 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3841 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3901 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     3961 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4021 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4081 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4141 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4201 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4261 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4321 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4381 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4441 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     4501 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     4561 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     4621 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     4681 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg a
//
