LOCUS       sequence-479472-        4879 bp    DNA     circular SYN 23-JUL-2025
DEFINITION  Tag S. pombe genes at 3' end (C-terminal end of protein) with 
            TagRFP-T, codon-optimized for S. pombe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4879)
  TITLE     Hauf Lab Plasmids 2025
REFERENCE   2  (bases 1 to 4879)
  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-pTagRFP-T-kanMX6
FEATURES             Location/Qualifiers
     source          1..4879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      792..979
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(847..871)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1054..2410
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1054..1397
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1398..2207
                     /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(1465..1484)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2213..2410
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2514..2533)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2515..2533)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2620..2637)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2791..3379)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2871..2890)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3550..4341,4342..4410))
                     /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     4173..4192
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4411..4515)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4583..4601
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4639..4661)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4761..4780
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4861..4879
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4861..4878
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gtttctaagg gtgaagaatt gattaaggaa aacatgcata tgaagcttta catggaagga
      121 acagttaaca accatcattt taagtgtact tctgaaggag aaggtaaacc ttacgaaggt
      181 actcaaacaa tgagaattaa ggttgttgaa ggtggacctc ttccatttgc ttttgatatt
      241 ttggcaactt cttttatgta cggatcacga acttttatta accatacaca aggtattcct
      301 gatttcttta agcaatcttt tccagaaggt tttacttggg aacgtgttac tacatatgaa
      361 gatggtggag ttttaactgc tacacaagat acatctttgc aagatggttg tcttatctat
      421 aacgttaaga ttagaggagt taattttcct tcaaacggtc cagttatgca aaagaaaact
      481 cttggatggg aagctaacac agaaatgtta tatcctgcag atggtggact tgaaggtcga
      541 actgatatgg ctttaaaact tgttggtgga ggtcatttga tttgtaactt taagactaca
      601 tatcgttcta aaaagcctgc taaaaacttg aagatgccag gagtttacta cgttgatcat
      661 cgtcttgaaa gaattaaaga agcagataag gaaacttacg ttgaacaaca tgaagttgct
      721 gttgcacgtt actgcgattt gccatcaaaa ttaggtcata agttgaatta aggcgcgcca
      781 cttctaaata agcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa
      841 aaataagtgt atacaaattt taaagtgact cttaggtttt aaaacgaaaa ttcttattct
      901 tgagtaactc tttcctgtag gtcaggttgc tttctcaggt atagtatgag gtcgctctta
      961 ttgaccacac ctctaccggc agatccgcta gggataacag ggtaatatag atctgtttag
     1021 cttgcctcgt ccccgccggg tcacccggcc agcgacatgg aggcccagaa taccctcctt
     1081 gacagtcttg acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta catttagccc
     1141 atacatcccc atgtataatc atttgcatcc atacattttg atggccgcac ggcgcgaagc
     1201 aaaaattacg gctcctcgct gcagacctgc gagcagggaa acgctcccct cacagacgcg
     1261 ttgaattgtc cccacgccgc gcccctgtag agaaatataa aaggttagga tttgccactg
     1321 aggttcttct ttcatatact tccttttaaa atcttgctag gatacagttc tcacatcaca
     1381 tccgaacata aacaaccatg ggtaaggaaa agactcacgt ttcgaggccg cgattaaatt
     1441 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     1501 gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     1561 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     1621 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     1681 tcaccactgc gatccccggc aaaacagcat tccaggtatt agaagaatat cctgattcag
     1741 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     1801 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga
     1861 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     1921 aagtctggaa agaaatgcat aagcttttgc cattctcacc ggattcagtc gtcactcatg
     1981 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     2041 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     2101 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     2161 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gtactgacaa
     2221 taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt
     2281 tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc
     2341 agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt
     2401 cgctatactg ctgtcgattc gatactaacg ccgccatcca gtttaaacga gctcgaattc
     2461 atcgatgata tcagatccac tagtggccta tgcggccgcg gatctgccgg tctccctata
     2521 gtgagtcgta ttaatttcga taagccaggt taacctgcat taatgaatcg gccaacgcgc
     2581 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2641 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2701 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2761 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2821 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2881 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2941 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3001 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3061 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3121 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3181 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3241 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3301 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3361 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3421 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3481 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3541 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3601 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3661 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3721 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3781 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3841 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3901 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3961 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4021 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4081 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4141 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4201 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4261 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4321 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4381 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4441 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4501 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     4561 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     4621 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     4681 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     4741 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgga
     4801 catattgtcg ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg
     4861 atttaggtga cactataga
//
