LOCUS       sequence-172996-        4715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tag S. pombe genes in C terminal with CLIP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4715)
  TITLE     pFA6a-mEGFP-KanMX6
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4715)
  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            "Unpublished"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-CLIP-KanMX6
FEATURES             Location/Qualifiers
     source          1..4715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             63..605
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylcytosine 
                     derivatives"
                     /label=CLIP-tag(TM)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="DKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVEV
                     PAPAAVLGGPEPLIQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLKV
                     VKFGEVISESHLAALVGNPAATAAVNTALDGNPVPILIPCHRVVQGDSDVGPYLGGLAV
                     KEWLLAHEGHRLGKPGLG"
     terminator      629..816
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(684..708)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            891..2247
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        891..1234
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1235..2044
                     /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(1302..1321)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2050..2247
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2351..2370)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2352..2370)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2457..2474)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2628..3216)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2708..2727)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3388..4179,4180..4248))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGSRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4011..4030
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4249..4353)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4421..4439
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4477..4499)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4599..4618
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4699..4715,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4699..4715,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acgacaaaga ctgcgaaatg aagcgcacca ccctggatag ccctctgggc aagctggaac
      121 tgtctgggtg cgaacagggc ctgcaccgta tcatcttcct gggcaaagga acatctgccg
      181 ccgacgccgt ggaagtgcct gccccagccg ccgtgctggg cggaccagag ccactgatcc
      241 aggccaccgc ctggctcaac gcctactttc accagcctga ggccatcgag gagttccctg
      301 tgccagccct gcaccaccca gtgttccagc aggagagctt tacccgccag gtgctgtgga
      361 aactgctgaa agtggtgaag ttcggagagg tcatcagcga gagccacctg gccgccctgg
      421 tgggcaatcc cgccgccacc gccgccgtga acaccgccct ggacggaaat cccgtgccca
      481 ttctgatccc ctgccaccgg gtggtgcagg gcgacagcga cgtggggccc tacctgggcg
      541 ggctcgccgt gaaagagtgg ctgctggccc acgagggcca cagactgggc aagcctgggc
      601 tgggttaggg cgcgccactt ctaaataagc gaatttctta tgatttatga tttttattat
      661 taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa
      721 acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata
      781 gtattaggtc gctcttattg accacacctc taccggcaga tccgctaggg ataacagggt
      841 aatatagatc tgtttagctt gcctcgtccc cgccgggtca cccggccagc gacatggagg
      901 cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc
      961 gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata cattttgatg
     1021 gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgctag cagggaaacg
     1081 ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag
     1141 gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat
     1201 acagttctca catcacatcc gaacataaac aaccatgggt aaggaaaaga ctcacgtttc
     1261 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     1321 taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga
     1381 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     1441 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     1501 tgatgatgca tggttactca ccactgcgat ccccggcaaa acagcattcc aggtattaga
     1561 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     1621 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     1681 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     1741 tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat tctcaccgga
     1801 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     1861 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     1921 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     1981 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     2041 ctaatcagta ctgacaataa aaagattctt gttttcaaga acttgtcatt tgtatagttt
     2101 ttttatattg tagttgttct attttaatca aatgttagcg tgatttatat tttttttcgc
     2161 ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat
     2221 cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat actaacgccg ccatccagtt
     2281 taaacgagct cgaattcatc gatgatatca gatccactag tggcctatgc ggccgcggat
     2341 ctgccggtct ccctatagtg agtcgtatta atttcgataa gccaggttaa cctgcattaa
     2401 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2461 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2521 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2581 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2641 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2701 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2761 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2821 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2881 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2941 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3001 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3061 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3121 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3181 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3241 gggtcttacg ctymagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3301 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3361 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3421 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3481 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3541 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3601 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3661 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3721 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3781 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3841 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3901 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3961 agaatagtgt atgcggctac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4021 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4081 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4141 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4201 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4261 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4321 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     4381 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     4441 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     4501 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     4561 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     4621 gagagtgcac catatggaca tattgtcgtt agaacgcggc tacaattaat acataacctt
     4681 atgtatcata cacatacgat ttaggtgaca ctata
//
