LOCUS       sequence-425144-        8626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A pLys3 backbone integrating plasmid expressing mECitrine under the 
            inducible promoter 41nmt1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8626)
  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 8626)
  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: pDB4881
FEATURES             Location/Qualifiers
     source          1..8626
                     /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"
     primer_bind     complement(4398..4426)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5021..5041
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      5109..5296
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5164..5188)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        5365..5708
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      6107..6304
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(6366..6386)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6366..6384)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6405..6421)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6405..6421)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6418..6440)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6429..6445
                     /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(6453..6459,6460..6477,6478..6483))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6498..6519
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6636..6653)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6807..7395)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6887..6906)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7566..8357,8358..8426))
                     /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     8189..8208
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8427..8531)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8599..8617
                     /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 agtaaaggag
     4381 aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc
     4441 acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta
     4501 aatttatttg cactactgga aaactacctg ttccatggcc aacacttgtc actactttgg
     4561 gttatggtct aatgtgcttt gctagatacc cagatcatat gaaacggcat gactttttca
     4621 agagtgccat gcccgaaggt tatgtacagg aaagaactat atttttcaaa gatgacggga
     4681 actacaagac acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat agaatcgagt
     4741 taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg gaatacaact
     4801 ataactcaca caatgtatac atcatggcag acaaacaaaa gaatggaatc aaagttaact
     4861 tcaaaattag acacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa
     4921 atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg tcctatcaat
     4981 ctaagctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa
     5041 cagctgctgg gattacacat ggcatggatg aactatacaa aagatcttaa cgcgccactt
     5101 ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa
     5161 taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga
     5221 gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc gctcttattg
     5281 accacacctc taccggcaga tccgctaggg ataacagggt aatatgttta gcttgcctcg
     5341 tccccgccgg gtcacccggc cagcgacatg gaggcccaga ataccctcct tgacagtctt
     5401 gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt acatttagcc catacatccc
     5461 catgtataat catttgcatc catacatttt gatggccgca cggcgcgaag caaaaattac
     5521 ggctcctcgc tgcagacctg cgagcaggga aacgctcccc tcacagacgc gttgaattgt
     5581 ccccacgccg cgcccctgta gagaaatata aaaggttagg atttgccact gaggttcttc
     5641 tttcatatac ttccttttaa aatcttgcta ggatacagtt ctcacatcac atccgaacat
     5701 aaacaaccat gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa
     5761 tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg
     5821 gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg
     5881 tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg
     5941 gaaatgagaa caggggcatc ttgagcccct gcggacggtg tcgacaggtg cttctcgatc
     6001 tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga
     6061 ttcgtgaatt gctgccctct ggttatgtgt gggagggcta atcagtactg acaataaaaa
     6121 gattcttgtt ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt
     6181 ttaatcaaat gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc
     6241 gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat
     6301 actgctgtcg attcgatact aacgccgcca tccacctgca ggtaatcacg tgccagcttt
     6361 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     6421 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6481 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6541 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6601 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6661 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6721 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6781 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6841 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6901 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6961 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7021 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7081 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7141 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7201 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     7261 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7321 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7381 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7441 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7501 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7561 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7621 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7681 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7741 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7801 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7861 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7921 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7981 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8041 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8101 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8161 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8221 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8281 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8341 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8401 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8461 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8521 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     8581 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
