LOCUS       sequence-247466-        4888 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 4888)
  TITLE     mNeonGreen tagging tools for yeast
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4888)
  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            "Unpublished"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-KanMX6
FEATURES             Location/Qualifiers
     source          1..4888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     104..122
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(160..182)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     282..301
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        382..400
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     382..399
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      1201..1388
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1256..1280)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1463..2819
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1463..1806
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1807..2616
                     /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     complement(1874..1893)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2622..2819
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2923..2942)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2924..2942)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3029..3046)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3200..3788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3280..3299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3959..4750,4751..4819))
                     /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     4582..4601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4820..4888,1..36))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
       61 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
      121 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
      181 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
      241 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
      301 actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac
      361 cttatgtatc atacacatac gatttaggtg acactataga acgcggccgc cagctgaagc
      421 ttcgtacgct gcaggtcgac ggatcggtga cggtgctggt ttaattaaca tggtgagtaa
      481 aggcgaggag gacaatatgg cgagcttacc tgctacccac gaactgcaca ttttcggctc
      541 aatcaatggt gtagattttg acatggtcgg tcaggggact ggtaacccta acgatggtta
      601 tgaggagcta aatctgaaat ctactaaagg ggaccttcaa tttagtccct ggatacttgt
      661 tccgcacata ggttatggtt tccaccaata tttgccctac cctgacggga tgagcccctt
      721 ccaggcggct atggtagatg gcagtggcta tcaggtccac agaacaatgc aatttgagga
      781 cggggccagc ctaaccgtga attataggta cacgtacgag gggtcacata taaagggcga
      841 agcacaggtg aaggggacgg ggttcccggc ggacggacca gtgatgacaa actccctgac
      901 cgctgccgat tggtgccgta gtaagaagac ttatcctaac gacaaaacga taatcagtac
      961 cttcaagtgg agttatacca cgggcaacgg gaaaagatac cgtagcacgg cgagaacaac
     1021 ttatactttc gccaagccaa tggctgccaa ctacttgaag aatcagccca tgtatgtatt
     1081 ccgtaagacg gagctgaaac atagcaaaac cgaactaaac ttcaaggagt ggcagaaagc
     1141 gtttaccgat gtgatgggta tggacgaatt gtataagtaa ggcgcgccac ttctaaataa
     1201 gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa aataagtgta
     1261 tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt gagtaactct
     1321 ttcctgtagg tcaggttgct ttctcaggta tagtatgagg tcgctcttat tgaccacacc
     1381 tctaccggca gatccgctag ggataacagg gtaatataga tctgtttagc ttgcctcgtc
     1441 cccgccgggt cacccggcca gcgacatgga ggcccagaat accctccttg acagtcttga
     1501 cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac atttagccca tacatcccca
     1561 tgtataatca tttgcatcca tacattttga tggccgcacg gcgcgaagca aaaattacgg
     1621 ctcctcgctg cagacctgcg agcagggaaa cgctcccctc acagacgcgt tgaattgtcc
     1681 ccacgccgcg cccctgtaga gaaatataaa aggttaggat ttgccactga ggttcttctt
     1741 tcatatactt ccttttaaaa tcttgctagg atacagttct cacatcacat ccgaacataa
     1801 acaaccatgg gtaaggaaaa gactcacgtt tcgaggccgc gattaaattc caacatggat
     1861 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     1921 tatcgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     1981 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     2041 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     2101 atccccggca aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     2161 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     2221 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     2281 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     2341 gaaatgcata agcttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     2401 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     2461 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     2521 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     2581 ttgcagtttc atttgatgct cgatgagttt ttctaatcag tactgacaat aaaaagattc
     2641 ttgttttcaa gaacttgtca tttgtatagt ttttttatat tgtagttgtt ctattttaat
     2701 caaatgttag cgtgatttat attttttttc gcctcgacat catctgccca gatgcgaagt
     2761 taagtgcgca gaaagtaata tcatgcgtca atcgtatgtg aatgctggtc gctatactgc
     2821 tgtcgattcg atactaacgc cgccatccag tttaaacgag ctcgaattca tcgatgatat
     2881 cagatccact agtggcctat gcggccgcgg atctgccggt ctccctatag tgagtcgtat
     2941 taatttcgat aagccaggtt aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3001 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3061 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3121 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3181 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3241 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3301 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3361 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     3421 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3481 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3541 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3601 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     3661 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3721 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3781 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3841 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3901 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3961 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4021 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4081 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4141 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4201 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4261 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4321 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4381 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4441 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4501 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4561 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4621 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4681 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4741 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4801 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4861 attgtctcat gagcggatac atatttga
//
