LOCUS       sequence-425123-        8775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A pHis5 backbone integrating plasmid expressing mTurquoise2 under 
            the inducible promoter 41nmt1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8775)
  AUTHORS   Pan ZQ, Yang YS, Du LL
  TITLE     Pil1 Co-tethering Assay to Detect Protein-Protein Interactions in 
            the Fission Yeast Schizosaccharomyces pombe.
  JOURNAL   Methods Mol Biol. 2025;2862:93-102. doi: 
            10.1007/978-1-0716-4168-2_7.
  PUBMED    39527195
REFERENCE   2  (bases 1 to 8775)
  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: "Methods Mol
            Biol."; date: "2025"; volume: "2862:93-102. doi"; pages: " 
            10.1007/978-1-0716-4168-2_7"
COMMENT     Sequence Label: pDB4896
FEATURES             Location/Qualifiers
     source          1..8775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             1188..1838
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQK
                     GEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQA
                     MGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLYS
                     FSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     rep_origin      complement(2272..2727)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2409..2430)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2621..2640
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2853..2875
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2867..2884
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2868..2884
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2894..2913
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2894..2912
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      4847..5034
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4902..4926)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            5102..6458
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        5102..5445
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             5446..6255
                     /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(5513..5532)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      6261..6458
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(6515..6535)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6515..6533)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6554..6570)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6554..6570)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6567..6589)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6578..6594
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6602..6608,6609..6626,6627..6632))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6647..6668
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6785..6802)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6956..7544)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7036..7055)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7715..8506,8507..8575))
                     /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     8338..8357
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8576..8680)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8748..8766
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaccg ggatggaacg attgaatact gtgctactct ttatgtaata taacgaattc
      241 aactgttcaa cgtatgtgac tgctctctgc ttctcggtat atatttactt gaaggaatta
      301 atttttcttt gccaatggtc tcttttacca attgcagtct ggcttaaagc aatgaggttg
      361 taattaaatg gcttgagacc tagaacaaac accttttaca atccttgttt actggttttg
      421 atagtaaatt cggttcgctt gtggtgtaca ttatattttt taggctagtc gtgatttttt
      481 gataatatca atattcttag atttgagaca ttcatgcttt ttaatttgat caatcgtttg
      541 tatcttcatt aattagtagg tggtttgata gaacatttag ttgcttgttt gactacaaat
      601 atctgcagaa tggattcggt taataaaaaa aatgtttgag gaattaattt tcgctttgga
      661 gtggaggagc ggccgctcct caattcgatg ttgcagattt tacaagtttt taaaatgtat
      721 ttcattatta ctttttatat gcctaataaa aaagccatag tttaatctat agataacttt
      781 ttttccagtg cactaacgga cgttacattc ccatacaaaa ctgcgtagtt aaagctaagg
      841 aaaagttaat atcatgttaa ttaaatacgc tatttacaat aagacattga actcatttat
      901 atcgttgaat atgaataacc aatttcagcg aatttttaac aaacatcgtt cacctcgttt
      961 aaggatatct tgtgtatggg gtgttgactt gctttatcga ataattaccg tacctgtaat
     1021 tggcttgctg gatatagcgg tagtctaata tctagcaaaa atcttttggg tgaaaaggct
     1081 tgcaatttca cgacaccgaa ctatttgtca ttttttaata aggaagtttt ccataaattc
     1141 ctgtaattct cggttgatct aattgaaaag agtagttttg catcacgatg aggagggctt
     1201 ttgtagaaag aaatacgaac gaaacgaaaa tcagcgttgc catcgctttg gacaaagctc
     1261 ccttacctga agagtcgaat tttattgatg aacttataac ttccaagcat gcaaaccaaa
     1321 agggagaaca agtaatccaa gtagacacgg gaattggatt cttggatcac atgtatcatg
     1381 cactggctaa acatgcaggc tggagcttac gactttactc aagaggtgat ttaatcatcg
     1441 atgatcatca cactgcagaa gatactgcta ttgcacttgg tattgcattc aagcaggcta
     1501 tgggtaactt tgccggcgtt aaaagatttg gacatgctta ttgtccactt gacgaagctc
     1561 tttctagaag cgtagttgac ttgtcgggac ggccctatgc tgttatcgat ttgggattaa
     1621 agcgtgaaaa ggttggggaa ttgtcctgtg aaatgatccc tcacttacta tattcctttt
     1681 cggtagcagc tggaattact ttgcatgtta cctgcttata tggtagtaat gaccatcatc
     1741 gtgctgaaag cgcttttaaa tctctggctg ttgccatgcg cgcggctact agtcttactg
     1801 gaagttctga agtcccaagc acgaagggag tgttgtaaag atgaattgga ttatgtcagg
     1861 aaaagaacga caattttgca tccaaattgt ctaaatttta gagttgcttg aaaacaatag
     1921 aaccttactt gctttataat tacgttaatt agaagcgtta tctcgtgaag gaatatagta
     1981 cgtagccgta taaattgaat tgaatgttca gcttatagaa tagagacact ttgctgttca
     2041 atgcgtcgtc acttaccata ctcactttat tatacgactt taagtataaa ctccgcggtt
     2101 atggtaaaat taatgatgca caaacgtccg attccatatg ggtacactac aattaaatac
     2161 ttttaagtaa cgctgatttc ttatttttta tccttggtgt atttattgtt tataccgcta
     2221 gggcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa
     2281 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     2341 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     2401 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     2461 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     2521 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     2581 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     2641 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2701 cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc aggctgcgca actgttggga
     2761 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     2821 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     2881 cagtgagcgc gcgtaatacg actcactata gggcgaattg ggtaccgggc ccgatcagaa
     2941 aattatcgcc ataaaagaca gaataagtca tcagcggttg tttcatttcc tatatttttt
     3001 ttttattttt ttatttttta ataagggaaa atttaacgtc taaggataca gaagattgtt
     3061 agcacattaa agtaataaag gcttaagtag taagtgcctt agcatgttat tgtatttcaa
     3121 aggacataat ctaaaataat aacaatatca tttctcacaa gttattcaat tttctttttt
     3181 tttctaataa tatcaagaat gtattatttg tttgacataa gtcaactaat ttatttaata
     3241 tgctggatta atcttgcaga catgtaaatt aacaagtttt agtcaaataa cgttgaagtt
     3301 tcaatgaact caaataattt ctcttttttt ttatataacc ataatctgat ttatattttc
     3361 cgcagggatc aactgaagtt atgacatttg gattggatca cttataacct tggtcgccaa
     3421 ataatacaaa aatcagcgtt ataaaacaaa gaaggttttt gttaagaaat taatcctctt
     3481 tcttgataag aaagttgaac cgaaattgca gatactgata tatgaaaata atacccacaa
     3541 ttttgggaat agcgcaagcc tcaatttaaa caataggtga ggacacatga taatgacctc
     3601 aatgattgtt agaagaaaag agcctcatta caaaatcgaa aaatgaatgg ttgggtacaa
     3661 gtttccaaaa catggtaaag tggactttgc gtatgagacg taaatagaaa aaaacacttg
     3721 ttatatgttt tctagaatta ttgttgtctc tttatggttg gatgatgcaa aatagtaatt
     3781 tcggttagtt gctgtaaaac accacgagac aaatagatat ggatatttat taaatcagga
     3841 aaaacgtaac tctcggctac tggatggttc agtcacccaa cgattactgg ggagagaaaa
     3901 cagggcaaaa gcaaagctta aaggaatccg attgtcattc ggcaatgtgc agcgaaacta
     3961 aaaaccggat aatggacctg ttaatcgaaa cattgaagat aaaggaagag gaatcctggc
     4021 atatcatcaa ttgaataagt tgaattaatt atttcaatct cattctcact ttctgactta
     4081 tagtcgcttt gttaaagcta gcatggtttc taagggtgaa gaactcttta ctggtgtcgt
     4141 tcctatcttg gttgaacttg atggagatgt taatggacac aagttttcag tctctggtga
     4201 aggagaaggt gatgctactt atggtaagct tacccttaaa ttcatttgca ctactggaaa
     4261 acttcctgtc ccttggccta ctcttgtcac taccctgtct tggggtgttc aatgctttgc
     4321 tcgttatcct gaccatatga agcaacacga tttctttaag agtgctatgc ctgaaggtta
     4381 tgttcaagag cgtactattt tcttcaaaga cgatggtaac tataagactc gtgctgaagt
     4441 taaatttgag ggtgatactt tagttaaccg tattgagctc aaaggaattg attttaaaga
     4501 ggatggtaat attcttggtc ataaattaga atacaattac ttttctgata atgtttacat
     4561 tactgccgat aaacaaaaga atggtatcaa ggctaacttc aagattcgtc ataacattga
     4621 agatggcgga gtacaacttg ctgatcatta ccaacaaaac actcctattg gtgacggtcc
     4681 tgttttgttg cctgataacc attatctttc tactcaatcc aaattgtcta aagatcctaa
     4741 tgaaaaacgt gatcacatgg tccttttgga atttgttacc gctgccggta ttaccttggg
     4801 tatggatgaa ttgtacaaga gatcttaacg cgccacttct aaataagcga atttcttatg
     4861 atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag
     4921 tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc tgtaggtcag
     4981 gttgctttct caggtatagt atgaggtcgc tcttattgac cacacctcta ccggcagatc
     5041 cgctagggat aacagggtaa tatgttagct tgcctcgtcc ccgccgggtc acccggccag
     5101 cgacatggag gcccagaata ccctccttga cagtcttgac gtgcgcagct caggggcatg
     5161 atgtgactgt cgcccgtaca tttagcccat acatccccat gtataatcat ttgcatccat
     5221 acattttgat ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc agacctgcga
     5281 gcagggaaac gctcccctca cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag
     5341 aaatataaaa ggttaggatt tgccactgag gttcttcttt catatacttc cttttaaaat
     5401 cttgctagga tacagttctc acatcacatc cgaacataaa caaccatggg taaggaaaag
     5461 actcacgttt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     5521 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc
     5581 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     5641 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     5701 atccgtactc ctgatgatgc atggttactc accactgcga tccccggcaa aacagcattc
     5761 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     5821 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     5881 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     5941 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa gcttttgcca
     6001 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     6061 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     6121 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     6181 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     6241 gatgagtttt tctaatcagt actgacaata aaaagattct tgttttcaag aacttgtcat
     6301 ttgtatagtt tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata
     6361 ttttttttcg cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat
     6421 catgcgtcaa tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc
     6481 gccatccagt ttaaacgagc tccagctttt gttcccttta gtgagggtta attgcgcgct
     6541 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     6601 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6661 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6721 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6781 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6841 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6901 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6961 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     7021 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     7081 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     7141 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     7201 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     7261 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7321 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7381 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7441 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     7501 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     7561 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     7621 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     7681 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     7741 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     7801 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     7861 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7921 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7981 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     8041 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     8101 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     8161 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     8221 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     8281 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     8341 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     8401 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     8461 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     8521 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     8581 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     8641 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     8701 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     8761 cgaggccctt tcgtc
//
