LOCUS       sequence-479477-        5108 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 5108)
  TITLE     Hauf Lab Plasmids 2025
REFERENCE   2  (bases 1 to 5108)
  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-hphNT1
FEATURES             Location/Qualifiers
     source          1..5108
                     /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"
     promoter        1054..1397
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1398..2426
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             1398..2426
                     /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      2434..2681
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2450..2468)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2743..2762)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2744..2762)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2849..2866)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3020..3608)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3100..3119)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3779..4570,4571..4639))
                     /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     4402..4421
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4640..4744)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4812..4830
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4868..4890)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4990..5009
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5090..5108
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5090..5107
                     /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 gcggacctgc gagcagggaa acgctcccct cacagacgcg
     1261 ttgaattgtc cccacgccgc gcccctgtag agaaatataa aaggttagga tttgccactg
     1321 aggttcttct ttcatatact tccttttaaa atcttgctag gatacagttc tcacatcaca
     1381 tccgaacata aacaaccatg ggtaaaaagc ctgaactcac cgcgacgtct gtcgagaagt
     1441 ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat
     1501 ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg
     1561 ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga
     1621 ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc
     1681 gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc
     1741 cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg
     1801 gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga
     1861 ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg
     1921 tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc
     1981 tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg
     2041 tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct
     2101 tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc
     2161 cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac
     2221 tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg
     2281 acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg
     2341 cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca
     2401 gcactcgtcc gagggcaaag gaataatctc gagtcatgta attagttatg tcacgcttac
     2461 attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag
     2521 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc
     2581 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
     2641 tgcttgagaa ggttttggga cgctcgaagg ctttaacgag ctcgaattca tcgatgatat
     2701 cagatccact agtggcctat gcggccgcgg atctgccggt ctccctatag tgagtcgtat
     2761 taatttcgat aagccaggtt aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2821 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2881 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2941 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3001 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3061 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3121 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3181 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3241 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3301 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3361 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3421 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3481 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3541 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3601 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3661 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3721 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3781 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3841 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3901 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3961 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4021 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4081 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4141 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4201 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4261 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4321 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4381 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4441 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4501 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4561 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4621 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4681 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4741 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     4801 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     4861 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     4921 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     4981 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatggac atattgtcgt
     5041 tagaacgcgg ctacaattaa tacataacct tatgtatcat acacatacga tttaggtgac
     5101 actataga
//
