LOCUS       sequence-425126-        8629 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A pLys3 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 8629)
  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 8629)
  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: pDB4893
FEATURES             Location/Qualifiers
     source          1..8629
                     /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"
     rep_origin      complement(2536..2991)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2673..2694)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2885..2904
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3117..3139
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3131..3148
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3132..3148
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3158..3177
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3158..3176
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      5112..5299
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5167..5191)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        5368..5711
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      6110..6307
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(6369..6389)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6369..6387)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6408..6424)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6408..6424)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6421..6443)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6432..6448
                     /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(6456..6462,6463..6480,6481..6486))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6501..6522
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6639..6656)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6810..7398)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6890..6909)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7569..8360,8361..8429))
                     /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     8192..8211
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8430..8534)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8602..8620
                     /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 gggccttacg aagttaaatt ataggcacag acatctttgg cattggaaag
      241 atttagtaac aatcataatg tagcacttct taaaaaatct aattttagaa caaaaaggct
      301 taaaaaagaa tggttccgct cttttaaaaa atagtaaaat gtacaatagc aaggttttcg
      361 taggtcgacg gacataacaa ggggaaagag aaataaagca tgtaaacgtc aataaaacat
      421 ctttgattct ggatcaatca tatgccaaat gaagaaaaat agcatttata acaacatgaa
      481 accatatttg agacctttta acgatgaaaa tttattaaat aaaagataaa aaagaaagaa
      541 accgttcaag ttcaatgaaa atttcgtcgt gaagtagaaa ccaagataat ggaaaggctt
      601 cgcaaagcca caaattttcc aatagtcatt ataactgaag attaaggaca gtaagcggcc
      661 gcttactagt aaaaatcaag tgaattgtgc ctaggatagg tagctaacgt gactgccact
      721 gaacctgtgt agcgtattag atccatcaag atcccttctg ctcatatcat tgctgatctg
      781 taaattgacc gcatcaaatt tgatctcatt tatatcgaga ttgatctata ttattttgcc
      841 aatttgcagt atataaactt atttgattaa gtttttggtc tccttcgcca cgtcttcatt
      901 ttgttagttt aaattttttt gactaactta gaaagggaag tttttaaaaa cgttgaaata
      961 ctttttcgat ttaaaaatat atatttaatt aattttctgt tccaaaatgg ttgctcctca
     1021 tctttggtta cgtgctgaaa ccaaaccttt agaagagcgc agtgctttga ctcctagaac
     1081 tgccaagata ctcgccgatg ctggatttca aattaccatt gagcgctctt cccaacgtgc
     1141 tttcaaggac aaagaatttg agcgccttgg tttccccatg gttcctgagg gctcatggcg
     1201 acatgcacct aaggacgcgt acattattgg tctcaaagag cttccggaaa atgataacag
     1261 tcctttgaag catactcata ttcaatttgc tcattgctac aaaaaccagg aaggttggcg
     1321 tgaagtcttg tcccgattcc ccgctggcaa tggtcttcta tatgatttgg aatttttgca
     1381 agatgacaat ggcagacgtg ttgccgcttt tggttaccac gctggttttg ctggatcagc
     1441 tatctcatgt ttagtttggg cacatcaatt attgcatcct aacaaacaat tccctgctat
     1501 ccgtccgttc cccaacgaaa aatctttggt tcgccatgtt gctcgtcaag tacgtttagc
     1561 tttgaaaaag aataacaacc agtatcctcg tattctggtt attggggctc ttggtcgttg
     1621 tggtactgga gcgtgcgatt tagcatcaaa gattggtatt cccttcgata acattcttcg
     1681 ctgggacatt aatgaaacta agaagggtgg tccctttacc gaaattacgg aatcagacat
     1741 atttgttaac tgcatttatc tgagcatgcc tattcccaaa ttctgtaccg tcgagtcgtt
     1801 aaatgtacct aaccgtaaat tgcgtgttgt ttgtgatgtt agctgtgaca ctaccaatcc
     1861 taacaatcct attccaattt ataatgtaaa cactacattt gatcatccta cagttgaggt
     1921 taaaggtgtc acaactcctc ctcctttaga ggtcatttct attgatcatc ttcctacttt
     1981 attgccaagg gaatcttccg aagcatttag tgaagctcta attcctagtc ttttggcttt
     2041 gaaggacgtc gacaatgctc ccgtttgggt cagagctaaa aaactttatg aaaccatggt
     2101 tcaaaagctt taaaataaat gttatttaat ttcaaagttc cgggtccttt atattgtttc
     2161 accaaattat tgaatattat tttctcaaaa attaacttta gaatacaatt tagacggcat
     2221 tttcgatgct ctggacattt tttccacgta cttgttaaat tgtagcggtg ttgcataata
     2281 ctaccttttg atttgttatt cacgttacaa tatatatttg actaatttaa atatgataat
     2341 atacacattt ctctgcaaac ctcctttata cttttcgccg gtcattgatt aataattaag
     2401 aaacaggatt tcatcctgca tttgccactt acctatatgg taaatataaa tcttgaaact
     2461 ttccccaata tttattgctt gctagggcgg tgtgaaatac cgcacagatg cgtaaggaga
     2521 aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     2581 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     2641 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2701 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2761 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     2821 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     2881 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     2941 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc
     3001 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     3061 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     3121 gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga
     3181 attgggtacc gggcccgatc agaaaattat cgccataaaa gacagaataa gtcatcagcg
     3241 gttgtttcat ttcctatatt ttttttttat ttttttattt tttaataagg gaaaatttaa
     3301 cgtctaagga tacagaagat tgttagcaca ttaaagtaat aaaggcttaa gtagtaagtg
     3361 ccttagcatg ttattgtatt tcaaaggaca taatctaaaa taataacaat atcatttctc
     3421 acaagttatt caattttctt ttttttttct aataatatca agaatgtatt atttgtttga
     3481 cataagtcaa ctaatttatt taatatgctg gattaatctt gcagacatgt aaattaacaa
     3541 gttttagtca aataacgttg aagtttcaat gaactcaaat aatttctctt tttttttata
     3601 taaccataat ctgatttata ttttccgcag ggatcaactg aagttatgac atttggattg
     3661 gatcacttat aaccttggtc gccaaataat acaaaaatca gcgttataaa acaaagaagg
     3721 tttttgttaa gaaattaatc ctctttcttg ataagaaagt tgaaccgaaa ttgcagatac
     3781 tgatatatga aaataatacc cacaattttg ggaatagcgc aagcctcaat ttaaacaata
     3841 ggtgaggaca catgataatg acctcaatga ttgttagaag aaaagagcct cattacaaaa
     3901 tcgaaaaatg aatggttggg tacaagtttc caaaacatgg taaagtggac tttgcgtatg
     3961 agacgtaaat agaaaaaaac acttgttata tgttttctag aattattgtt gtctctttat
     4021 ggttggatga tgcaaaatag taatttcggt tagttgctgt aaaacaccac gagacaaata
     4081 gatatggata tttattaaat caggaaaaac gtaactctcg gctactggat ggttcagtca
     4141 cccaacgatt actggggaga gaaaacaggg caaaagcaaa gcttaaagga atccgattgt
     4201 cattcggcaa tgtgcagcga aactaaaaac cggataatgg acctgttaat cgaaacattg
     4261 aagataaagg aagaggaatc ctggcatatc atcaattgaa taagttgaat taattatttc
     4321 aatctcattc tcactttctg acttatagtc gctttgttaa agctagcatg gtttctaagg
     4381 gtgaagaact ctttactggt gtcgttccta tcttggttga acttgatgga gatgttaatg
     4441 gacacaagtt ttcagtctct ggtgaaggag aaggtgatgc tacttatggt aagcttaccc
     4501 ttaaattcat ttgcactact ggaaaacttc ctgtcccttg gcctactctt gtcactaccc
     4561 tgtcttgggg tgttcaatgc tttgctcgtt atcctgacca tatgaagcaa cacgatttct
     4621 ttaagagtgc tatgcctgaa ggttatgttc aagagcgtac tattttcttc aaagacgatg
     4681 gtaactataa gactcgtgct gaagttaaat ttgagggtga tactttagtt aaccgtattg
     4741 agctcaaagg aattgatttt aaagaggatg gtaatattct tggtcataaa ttagaataca
     4801 attacttttc tgataatgtt tacattactg ccgataaaca aaagaatggt atcaaggcta
     4861 acttcaagat tcgtcataac attgaagatg gcggagtaca acttgctgat cattaccaac
     4921 aaaacactcc tattggtgac ggtcctgttt tgttgcctga taaccattat ctttctactc
     4981 aatccaaatt gtctaaagat cctaatgaaa aacgtgatca catggtcctt ttggaatttg
     5041 ttaccgctgc cggtattacc ttgggtatgg atgaattgta caagagatct taacgcgcca
     5101 cttctaaata agcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa
     5161 aaataagtgt atacaaattt taaagtgact cttaggtttt aaaacgaaaa ttcttattct
     5221 tgagtaactc tttcctgtag gtcaggttgc tttctcaggt atagtatgag gtcgctctta
     5281 ttgaccacac ctctaccggc agatccgcta gggataacag ggtaatacgt ttagcttgcc
     5341 tcgtccccgc cgggtcaccc ggccagcgac atggaggccc agaataccct ccttgacagt
     5401 cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc cgtacattta gcccatacat
     5461 ccccatgtat aatcatttgc atccatacat tttgatggcc gcacggcgcg aagcaaaaat
     5521 tacggctcct cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat
     5581 tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc actgaggttc
     5641 ttctttcata tacttccttt taaaatcttg ctaggataca gttctcacat cacatccgaa
     5701 cataaacaac catgcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta
     5761 caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg
     5821 acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac
     5881 tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga
     5941 tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgtcgacag gtgcttctcg
     6001 atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg
     6061 ggattcgtga attgctgccc tctggttatg tgtgggaggg ctaatcagta ctgacaataa
     6121 aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct
     6181 attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga
     6241 tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc
     6301 tatactgctg tcgattcgat actaacgccg ccatccacct gcaggtaatc acgtgccagc
     6361 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     6421 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6481 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6541 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6601 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6661 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6721 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6781 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6841 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6901 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6961 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     7021 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     7081 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     7141 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     7201 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     7261 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7321 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7381 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7441 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7501 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7561 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7621 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7681 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7741 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7801 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7861 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7921 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7981 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     8041 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     8101 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     8161 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     8221 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     8281 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     8341 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     8401 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     8461 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     8521 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     8581 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc
//
