LOCUS       sequence-479469-        5093 bp    DNA     circular SYN 23-JUL-2025
DEFINITION  Tag S. pombe genes at 3' end (C-terminal end of protein) with 
            mKate2, codon-optimized for S. pombe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5093)
  TITLE     Hauf Lab Plasmids 2025
REFERENCE   2  (bases 1 to 5093)
  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-pmKate2-hphNT1
FEATURES             Location/Qualifiers
     source          1..5093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      777..964
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(832..856)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1039..1382
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1383..2411
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             1383..2411
                     /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      2419..2666
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2435..2453)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2728..2747)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2729..2747)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2834..2851)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3005..3593)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3085..3104)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3764..4555,4556..4624))
                     /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     4387..4406
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4625..4729)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4797..4815
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4853..4875)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4975..4994
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5075..5093
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5075..5092
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gtttctgaat tgattaagga aaacatgcat atgaagcttt acatggaagg aacagttaac
      121 aaccatcatt ttaagtgtac atctgaagga gaaggtaaac cttacgaagg tactcaaaca
      181 atgagaatta aggctgttga aggtggacct ttgccatttg cttttgatat tcttgcaact
      241 tcttttatgt acggatcaaa gacttttatt aaccatacac aaggtattcc tgatttcttt
      301 aagcaatctt ttccagaagg ttttacttgg gaaagagtta ctacatatga agatggtgga
      361 gttttgactg ctacacaaga tacatcactt caagatggtt gtcttatcta taacgttaag
      421 attcgaggag ttaactttcc ttctaacggt ccagttatgc aaaagaaaac tttaggatgg
      481 gaagcttcaa ctgaaacact ttatcctgca gatggtggat tggaaggtag agctgatatg
      541 gcattaaaac ttgttggtgg aggtcatttg atttgtaatc ttaagactac ataccgatct
      601 aaaaagcctg ctaaaaactt aaagatgcca ggagtttact acgttgatcg tagacttgaa
      661 cgtattaaag aagcagataa ggaaacttac gttgaacaac atgaagttgc tgttgcacga
      721 tactgcgatc ttccatctaa attgggtcat cgttaaggcg cgccacttct aaataagcga
      781 atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca
      841 aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc
      901 tgtaggtcag gttgctttct caggtatagt atgaggtcgc tcttattgac cacacctcta
      961 ccggcagatc cgctagggat aacagggtaa tatagatctg tttagcttgc ctcgtccccg
     1021 ccgggtcacc cggccagcga catggaggcc cagaataccc tccttgacag tcttgacgtg
     1081 cgcagctcag gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta
     1141 taatcatttg catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc
     1201 tcgctgcgga cctgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac
     1261 gccgcgcccc tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat
     1321 atacttcctt ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa
     1381 ccatgggtaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt
     1441 tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct
     1501 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca
     1561 aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg
     1621 acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca
     1681 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca
     1741 tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc
     1801 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg
     1861 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg
     1921 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt
     1981 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg
     2041 aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt
     2101 tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat
     2161 cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg
     2221 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
     2281 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg
     2341 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg
     2401 caaaggaata atctcgagtc atgtaattag ttatgtcacg cttacattca cgccctcccc
     2461 ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt
     2521 atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     2581 ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt
     2641 tgggacgctc gaaggcttta acgagctcga attcatcgat gatatcagat ccactagtgg
     2701 cctatgcggc cgcggatctg ccggtctccc tatagtgagt cgtattaatt tcgataagcc
     2761 aggttaacct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2821 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2881 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2941 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3001 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3061 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3121 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3181 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3241 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3301 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3361 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3421 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3481 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3541 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3601 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3661 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3721 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3781 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3841 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3901 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3961 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4021 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4081 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4141 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4201 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4261 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4321 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4381 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4441 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4501 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4561 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4621 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4681 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4741 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     4801 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     4861 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     4921 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     4981 cagagcagat tgtactgaga gtgcaccata tggacatatt gtcgttagaa cgcggctaca
     5041 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat aga
//
