LOCUS       sequence-172998-        4861 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tag S. pombe genes in C terminal with PATagRFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4861)
  TITLE     pFA6a-mEGFP-KanMX6
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4861)
  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            "Unpublished"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-PATagRFP-KanMX6
FEATURES             Location/Qualifiers
     source          1..4861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      776..963
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(831..855)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1038..2394
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1038..1381
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1382..2191
                     /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(1449..1468)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2197..2394
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2498..2517)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2499..2517)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2604..2621)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2775..3363)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2855..2874)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3534..4325,4326..4394))
                     /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     4157..4176
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4395..4499)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4567..4585
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4623..4645)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4745..4764
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4845..4861,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4845..4861,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acagcgagct gattaaggag aacatgcaca tgaagctgta catggagggc accgtgaaca
      121 accaccactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
      181 tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacca
      241 gcttcatgta cggcagcagc accttcatca accacaccca gggcatcccc gacttctgga
      301 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg
      361 tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga
      421 tcagaggggt gaacttccca tccaacggcc ctgtgatgaa gaagaaaaca ctcggctggg
      481 aaccgagcac cgagaaactg aaacccgctg acggcggcct ggaaggcagg gtcgacatgg
      541 ccctgaagct cgtgggcggg ggccacctga tctgcaactt caagaccaca tacagatcca
      601 agaaacccgc taagaacctc aagatgcccg gcgtctacta tgtggaccgc agactggaaa
      661 tcatcaagga ggccgacaaa gagacctact gggagcagca cgaggtggct gtggccagat
      721 actccgacct ccctagcaaa ctggggcaca agtaaggcgc gccacttcta aataagcgaa
      781 tttcttatga tttatgattt ttattattaa ataagttata aaaaaaataa gtgtatacaa
      841 attttaaagt gactcttagg ttttaaaacg aaaattctta ttcttgagta actctttcct
      901 gtaggtcagg ttgctttctc aggtatagta tgaggtcgct cttattgacc acacctctac
      961 cggcagatcc gctagggata acagggtaat atagatctgt ttagcttgcc tcgtccccgc
     1021 cgggtcaccc ggccagcgac atggaggccc agaataccct ccttgacagt cttgacgtgc
     1081 gcagctcagg ggcatgatgt gactgtcgcc cgtacattta gcccatacat ccccatgtat
     1141 aatcatttgc atccatacat tttgatggcc gcacggcgcg aagcaaaaat tacggctcct
     1201 cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat tgtccccacg
     1261 ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc actgaggttc ttctttcata
     1321 tacttccttt taaaatcttg ctaggataca gttctcacat cacatccgaa cataaacaac
     1381 catgggtaag gaaaagactc acgtttcgag gccgcgatta aattccaaca tggatgctga
     1441 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     1501 attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     1561 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     1621 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     1681 cggcaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     1741 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     1801 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     1861 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     1921 gcataagctt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     1981 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     2041 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     2101 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     2161 gtttcatttg atgctcgatg agtttttcta atcagtactg acaataaaaa gattcttgtt
     2221 ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat
     2281 gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt
     2341 gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg
     2401 attcgatact aacgccgcca tccagtttaa acgagctcga attcatcgat gatatcagat
     2461 ccactagtgg cctatgcggc cgcggatctg ccggtctccc tatagtgagt cgtattaatt
     2521 tcgataagcc aggttaacct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2581 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2641 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2701 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2761 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2821 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2881 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2941 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3001 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3061 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3121 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3181 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3241 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3301 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3361 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3421 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3481 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3541 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3601 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3661 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3721 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3781 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3841 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3901 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3961 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4021 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4081 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4141 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4201 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4261 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4321 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4381 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4441 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     4501 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     4561 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     4621 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     4681 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     4741 tatgcggcat cagagcagat tgtactgaga gtgcaccata tggacatatt gtcgttagaa
     4801 cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat
     4861 a
//
