LOCUS       sequence-181132-        4565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  genomic GBP epitope C-terminal tagging in S. pombe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4565)
  AUTHORS   Chen YH, Wang GY, Hao HC, Chao CJ, Wang Y, Jin QW
  TITLE     Facile manipulation of protein localization in fission yeast through
            GBP-GFP binding.
  JOURNAL   J Cell Sci. 2017 Jan 12. pii: jcs.198457. doi: 10.1242/jcs.198457.
  PUBMED    28082423
REFERENCE   2  (bases 1 to 4565)
  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: "J Cell Sci.
            2017 Jan 12. pii: jcs.198457. doi: 10.1242/jcs.198457."
COMMENT     Sequence Label: pFA6a-GBP-kanMX6
FEATURES             Location/Qualifiers
     source          1..4565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             70..411
                     /codon_start=1
                     /product="GFP-binding fragment of a single-chain camelid 
                     antibody (Rothbauer et al., 2008)"
                     /label=GFP nanobody
                     /translation="VQLVESGGALVQPGGSLRLSCAASGFPVNRYSMRWYRQAPGKERE
                     WVAGMSSAGDRSSYEDSVKGRFTISRDDARNTVYLQMNSLKPEDTAVYYCNVNVGFEYW
                     GQGTQVTVSS"
     terminator      478..665
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(533..557)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            740..2096
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        740..1083
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1084..1893
                     /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(1151..1170)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1899..2096
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2200..2219)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2201..2219)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2306..2323)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2477..3065)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2557..2576)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3236..4027,4028..4096))
                     /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     3859..3878
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4097..4201)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4269..4287
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4325..4347)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4447..4466
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4547..4565
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4547..4564
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 atggctgatg tccaactggt ggagtctggt ggcgctttgg tgcagccagg tggctctctg
      121 cgtttgtcct gtgccgcttc tggcttccca gtgaaccgct attccatgcg ctggtatcgc
      181 caggctccag gcaaagagcg tgagtgggta gccggtatgt ccagcgcggg tgatcgtagc
      241 tcctatgaag actccgtgaa gggccgtttc accatcagcc gtgacgatgc ccgtaacacg
      301 gtgtatctgc aaatgaacag cttgaaacct gaagatacgg ccgtgtatta ctgtaatgtg
      361 aacgtgggct tcgagtattg gggccaaggc acccaggtca ccgtctcctc tggaaggtaa
      421 gcggccgctc gttccagatt acgctgctca gtgctgaggc gcgccacttc taaataagcg
      481 aatttcttat gatttatgat ttttattatt aaataagtta taaaaaaaat aagtgtatac
      541 aaattttaaa gtgactctta ggttttaaaa cgaaaattct tattcttgag taactctttc
      601 ctgtaggtca ggttgctttc tcaggtatag tatgaggtcg ctcttattga ccacacctct
      661 accggcagat ccgctaggga taacagggta atatagatct gtttagcttg cctcgtcccc
      721 gccgggtcac ccggccagcg acatggaggc ccagaatacc ctccttgaca gtcttgacgt
      781 gcgcagctca ggggcatgat gtgactgtcg cccgtacatt tagcccatac atccccatgt
      841 ataatcattt gcatccatac attttgatgg ccgcacggcg cgaagcaaaa attacggctc
      901 ctcgctgcag acctgcgagc agggaaacgc tcccctcaca gacgcgttga attgtcccca
      961 cgccgcgccc ctgtagagaa atataaaagg ttaggatttg ccactgaggt tcttctttca
     1021 tatacttcct tttaaaatct tgctaggata cagttctcac atcacatccg aacataaaca
     1081 accatgggta aggaaaagac tcacgtttcg aggccgcgat taaattccaa catggatgct
     1141 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     1201 cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     1261 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     1321 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     1381 cccggcaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     1441 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     1501 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     1561 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     1621 atgcataagc ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     1681 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     1741 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     1801 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     1861 cagtttcatt tgatgctcga tgagtttttc taatcagtac tgacaataaa aagattcttg
     1921 ttttcaagaa cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa
     1981 atgttagcgt gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa
     2041 gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt
     2101 cgattcgata ctaacgccgc catccagttt aaacgagctc gaattcatcg atgatatcag
     2161 atccactagt ggcctatgcg gccgcggatc tgccggtctc cctatagtga gtcgtattaa
     2221 tttcgataag ccaggttaac ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2281 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2341 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2401 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2461 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2521 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2581 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2641 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2701 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2761 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2821 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2881 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2941 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3001 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3061 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3121 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3181 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3241 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3301 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3361 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3421 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3481 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3541 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3601 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3661 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3721 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3781 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3841 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3901 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3961 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4021 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4081 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4141 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4201 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     4261 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     4321 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     4381 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     4441 actatgcggc atcagagcag attgtactga gagtgcacca tatggacata ttgtcgttag
     4501 aacgcggcta caattaatac ataaccttat gtatcataca catacgattt aggtgacact
     4561 ataga
//
