LOCUS       sequence-479468-        4876 bp    DNA     circular SYN 23-JUL-2025
DEFINITION  Tag S. pombe genes at 3' end (C-terminal end of protein) with 
            mNeonGreen, codon-optimized for S. pombe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4876)
  TITLE     Hauf Lab Plasmids 2025
REFERENCE   2  (bases 1 to 4876)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pFA6a-pmNeonGreen-kanMX6
FEATURES             Location/Qualifiers
     source          1..4876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      789..976
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(844..868)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1051..2407
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1051..1394
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1395..2204
                     /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(1462..1481)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2210..2407
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2511..2530)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2512..2530)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2617..2634)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2788..3376)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2868..2887)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3547..4338,4339..4407))
                     /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     4170..4189
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4408..4512)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4580..4598
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4636..4658)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4758..4777
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4858..4876
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4858..4875
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gtttctaagg gtgaagaaga taacatggca tcattgcctg ctacacatga attgcatatt
      121 tttggttcta ttaacggagt tgattttgat atggttggtc aaggaactgg taaccctaac
      181 gatggatacg aagaacttaa ccttaagtct acaaagggag atcttcaatt ttcaccttgg
      241 attcttgttc cacatattgg atacggtttt catcaatact taccttatcc agatggaatg
      301 tctccatttc aagctgcaat ggttgatgga tcaggttatc aagttcatcg aactatgcaa
      361 tttgaagatg gtgcttctct tactgttaac taccgttaca catacgaagg atcacatatt
      421 aaaggtgaag ctcaagttaa gggaactggt tttcctgctg atggtccagt tatgactaat
      481 tctttgacag ctgcagattg gtgtcgttct aagaaaactt atcctaacga taagactatt
      541 atttctactt ttaaatggtc atacactaca ggaaatggta aacgttatag atctactgct
      601 agaactacat acacatttgc aaaacctatg gctgcaaact acttaaagaa ccaaccaatg
      661 tacgtttttc gaaagactga acttaagcat tctaagacag aattgaactt taaggaatgg
      721 caaaaggctt ttactgatgt tatgggtatg gatgaacttt acaaataggg cgcgccactt
      781 ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa
      841 taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga
      901 gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc gctcttattg
      961 accacacctc taccggcaga tccgctaggg ataacagggt aatatagatc tgtttagctt
     1021 gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac
     1081 agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata
     1141 catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa
     1201 aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg
     1261 aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg
     1321 ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc
     1381 gaacataaac aaccatgggt aaggaaaaga ctcacgtttc gaggccgcga ttaaattcca
     1441 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     1501 cgacaatcta tcgattgtat gggaagcccg atgcgccaga gctgtttctg aaacatggca
     1561 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     1621 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     1681 ccactgcgat ccccggcaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     1741 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     1801 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     1861 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     1921 tctggaaaga aatgcataag cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     1981 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     2041 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     2101 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     2161 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagta ctgacaataa
     2221 aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct
     2281 attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga
     2341 tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc
     2401 tatactgctg tcgattcgat actaacgccg ccatccagtt taaacgagct cgaattcatc
     2461 gatgatatca gatccactag tggcctatgc ggccgcggat ctgccggtct ccctatagtg
     2521 agtcgtatta atttcgataa gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg
     2581 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2641 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2701 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2761 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2821 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2881 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2941 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3001 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3061 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3121 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3181 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3241 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3301 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3361 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3421 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3481 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3541 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3601 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3661 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3721 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3781 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3841 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3901 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3961 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4021 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4081 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4141 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4201 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4261 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4321 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4381 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4441 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4501 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     4561 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     4621 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4681 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4741 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatggacat
     4801 attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
     4861 taggtgacac tataga
//
