LOCUS       sequence-269073-        9005 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mTagBFP2 tagged CRIB domain to monitor active Cdc42.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9005)
  AUTHORS   Vjestica A, Marek M, Nkosi PJ, Merlini L, Liu G, Berard M, 
            Billault-Chaumartin I, Martin SG
  TITLE     A toolbox of stable integration vectors in the fission yeast 
            Schizosaccharomyces pombe.
  JOURNAL   J Cell Sci. 2020 Jan 8;133(1). pii: jcs.240754. doi: 
            10.1242/jcs.240754.
  PUBMED    31801797
REFERENCE   2  (bases 1 to 9005)
  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."; date: "2020-01-8"; volume: "133(1). pii: jcs.240754. doi"; 
            pages: " 10.1242/jcs.240754"
COMMENT     Sequence Label: pAV0816
FEATURES             Location/Qualifiers
     source          1..9005
                     /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(2697..3152)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2834..2855)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3046..3065
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3278..3300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3292..3309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3293..3309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3319..3338
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3319..3337
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4753..5463
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     terminator      5487..5674
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5542..5566)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        5749..6092
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      6491..6688
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(6745..6765)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6745..6763)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6784..6800)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6784..6800)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6797..6819)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6808..6824
                     /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(6832..6838,6839..6856,6857..6862))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6877..6898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7015..7032)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7186..7774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7266..7285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7945..8736,8737..8805))
                     /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     8568..8587
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8806..8910)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8978..8996
                     /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 ggagaatcct gaataatgtg ctgtgaagca gttgaaagaa aattaaagaa
      241 cccaatataa tatgctataa agcaatcaaa aaataatagt aagaaaccca aataaaaccc
      301 cgttgaccat gttcccaatt aaattaacgc atattaatgc aaaaaatgca atattaagac
      361 tagttattaa accgataatt agtcttcaat tattaaaaga tgtagttgat ttgtattatc
      421 tttcaattgc taattctaaa tatattggag aattcaatga gtttatgcat tgaagtttaa
      481 gataacattg gcgctgattg cagcaaaagt aatcacgcat atagctgtag gaccaaaacg
      541 atcatcatat cggaccgcat tatgtttaaa cgttgcttta tatatggata acgacatata
      601 catttatttc ttgaaaaaat agcaaagcta tattgatttt agcgtatgct gctttaaagt
      661 tgcatttcac aatgcttgga aatgtaacga tgacagaatt aatacgatgc aaaactcaaa
      721 taataaactg cgcactaact cactacaata aacaactttg aaaaaaagat tcgttttttc
      781 aacatttacc atctcattaa gctgagctgc caaggtatat acatacttca tcgaatatga
      841 gcgaaaaaca ggttgtaggg atccttggag gtggtcaatt gggccgaatg atggtagagg
      901 cagcccatcg cttaaacatc aaatgcatca tcttggatgc agcaaattct cctgccaaac
      961 aaattgatgg aggacgtgag cacattgatg catcatttac tgaccccgat gcaattgttg
     1021 aactgtctaa gaagtgcacg ttattaacaa ctgaaattga gcatattaac actgatgcct
     1081 tggcagccgt tacgaaatct gttgctgttg aaccctctcc tgcaactctg cgatgcattc
     1141 aagacaaata tcttcaaaaa cagcatttac aggtttttaa gattgcactt cctgaatttt
     1201 gcgatgcacc tgaccaggaa agtgttgaaa aagcaggcca agagtttggt tatccttttg
     1261 tactgaaaag taaaacattg gcttacgacg gtcgtggaaa ttacgttgtt catcaaccat
     1321 ctgagattcc tactgccatc aaagcacttg gtgatcgtcc gctttatgtt gaaaagttcg
     1381 ttcctttctc catggaaatt gcagtgatgg tagtacgcag tttagacgga aaagtttatg
     1441 cttatcctac aactgagacc attcaaaagg ataatgtttg tcatttagta tatgcccctg
     1501 ctcgtcttcc cttctcaatt caacagcgtg ctcaaaccct tgccatggat gcggtgcgca
     1561 cttttgaagg cgctggtata tatggtgtag agatgtttgt tttgcaagat ggtgagacca
     1621 ttttactcaa cgaaattgct cctcggcctc acaattcagg tcactacacc attgaagctt
     1681 gcccaacttc tcagtttgaa gctcacttac gggccatatg tggtcttcct ttcagcgaaa
     1741 tcaacaccca actctcgact tccacaactc atgcgttgat ggtaaatatt ttaggtactg
     1801 atgatcctga ttatgtttca aagatcgcta aacgttctct gtccattccc ggtgcaactt
     1861 tgcatcttta tggtaaagct gaatctagaa agggtcgcaa gatgggacac gttaccatca
     1921 tttctgattc acctcaagaa tgtgaacgta ggtatcagat gcttcttgac gtcaaagatc
     1981 ctgtcgaatc acctgttgtt ggtattatca tgggttcgga ttctgattta agcaagatga
     2041 aagatgctgc cgtcatttta gatgaattca aggtgcctta cgaacttact attgtttcag
     2101 ctcaccgcac accagatcgc atggttactt atgctcgtac cgcagcttca agagggttgc
     2161 gtgtgattat tgctggtgct ggtggtgccg ctcatttgcc tggtatggtt gctgcaatga
     2221 cacctcttcc agtaatcggc gttcctgtaa aaggaagcac tcttgacgga gttgactctc
     2281 ttcactctat tgttcagatg cctcgaggtg tccctgtcgc cactgttgct atcaataata
     2341 gccaaaatgc cggtatttta gcctgtcgta tacttgctac atttcaaccc tcccttttgg
     2401 ctgctatgga gagctttatg gacaatatga aagagattgt tttagaaaag gctgataaat
     2461 tagaaaaagt tggttggaaa aattattctg cataggcgac catagacata actgttaaat
     2521 gattttgctt gctttttgtt ttaaataata ttttattttg aaattctaga ttgtaaaatg
     2581 tattctaaaa agtgattaaa aattaaaaat aaaaaccccc cgaataatgt gctgcgacgc
     2641 agctagggcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat
     2701 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     2761 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     2821 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     2881 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     2941 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3001 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3061 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3121 cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt
     3181 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     3241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     3301 acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc
     3361 ctcgaggatc tacgataatg agacggtgtt tgggttgttt ttactgaatt tttcttttat
     3421 tttatgacat cttaattttt atgttaatga ttattcttat acttttatct atctttgatt
     3481 aaaactaata ttcattcttt tgatcactaa ttatttttat agtatacgat tcaacccaac
     3541 tataatacat tgtatcaaga gaaattttta attagatgta atttaatagc catttgataa
     3601 aaaaaaatta cctttttttt tgctagtgtt aaattattga tcataacaat gtcgaatgaa
     3661 tactaataaa atagggacac gcgagttgca aatacttgaa atacctttga agatttgtct
     3721 gtgaattttg ataagatttt tatgaccatc tcttttatat tcgatatcgt gaactttcat
     3781 tttcttttgt tttttgcttt tcaagttaaa taacgcggaa attatatttc cttatatgtt
     3841 tataagagct tttaaaggat ttgctatatc gcttacagta ctacaaccgg ttgctcaatg
     3901 ttatccgttt ccgcggtctt cttccgtgcg cattctgccg tgaagtgcta acgctgtgtg
     3961 tggctttatc aaaaaagtgg ctcaaatata gcaatttcat acatatttaa acacccatta
     4021 tacttaaatg tatattttaa aatatataag agattttcac tgtataactg tgtatcttat
     4081 ctatatgtaa tcaacggctt cataccacct cagccagccg tgttataact taccgtttac
     4141 caactacatt ttttgtaacg aaccaaaaaa ccctcaaaag acaagaccat gggaggtccc
     4201 gggactagtg caagtattac caatactgga aacgaaacca tgaaccttcc acagatgcgg
     4261 tcgatttggc tggatgaaga tgaagaagct gaaaaactct acggtctgca ggcccagcaa
     4321 ttcatgggat ctgatgatga agaaaaccta ggcattactt tcatcaacag cgataaacct
     4381 gtgctgagta acaagaaaaa cattgagttg cctccacttt caccaaattc acatccgtct
     4441 tgccaccaca ggagaagtaa ttctaactct gcaaagtcta aagaatcatc gtcatcatcg
     4501 tccagcgcca acaagacaaa tcacaaaaag gttttcctta agcttaattt gttgaagaaa
     4561 aagttgcttg gtgcccaacc ggacataaga ggtaaaggta tctccacacc atttgatttt
     4621 caacatattt cacatgctga cactagaaat ggattccaag atgagcaatt gcaggaacct
     4681 tcatcgctgt ccacagagat taaggacgac tatacctcct cctcaagcaa gcgggattcg
     4741 aaattaatta acatggtgtc taagggcgaa gagctgatta aggagaacat gcacatgaag
     4801 ctgtacatgg agggcaccgt ggacaaccat cacttcaagt gcacatccga gggcgaaggc
     4861 aagccctacg agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc
     4921 ttcgccttcg acatcctggc tactagcttc ctctacggca gcaagacctt catcaaccac
     4981 acccagggca tccccgactt cttcaagcag tccttccctg agggcttcac atgggagaga
     5041 gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac
     5101 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcacatccaa cggccctgtg
     5161 atgcagaaga aaacactcgg ctgggaggcc ttcaccgaga cgctgtaccc cgctgacggc
     5221 ggcctggaag gcagaaacga catggccctg aagctcgtgg gcgggagcca tctgatcgca
     5281 aacgccaaga ccacatatag atccaagaaa cccgctaaga acctcaagat gcctggcgtc
     5341 tactatgtgg actacagact ggaaagaatc aaggaggcca acaacgagac ctacgtcgag
     5401 cagcacgagg tggcagtggc cagatactgc gacctcccta gcaaactggg gcacaagctt
     5461 aactaaggcg cgccacttct aaataagcga atttcttatg atttatgatt tttattatta
     5521 aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac
     5581 gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagt
     5641 atgaggtcgc tcttattgac cacacctcta ccggcagatc cgctagggat aacagggtaa
     5701 tatagatctg tttagcttgc ctcgtccccg ccgggtcacc cggccagcga catggaggcc
     5761 cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc
     5821 ccgtacattt agcccataca tccccatgta taatcatttg catccataca ttttgatggc
     5881 cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca gggaaacgct
     5941 cccctcacag acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt
     6001 taggatttgc cactgaggtt cttctttcat atacttcctt ttaaaatctt gctaggatac
     6061 agttctcaca tcacatccga acataaacaa ccatgccttt gtctcaagaa gaatccaccc
     6121 tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg
     6181 ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta
     6241 ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca
     6301 acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac
     6361 ggtgtcgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg
     6421 atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg
     6481 gctaatcagt actgacaata aaaagattct tgttttcaag aacttgtcat ttgtatagtt
     6541 tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata ttttttttcg
     6601 cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa
     6661 tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc gccatccagt
     6721 ttaaacgagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     6781 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6841 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6901 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6961 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     7021 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     7081 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     7141 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     7201 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     7261 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     7321 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     7381 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7441 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7501 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7561 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7621 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7681 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7741 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     7801 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7861 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7921 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7981 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     8041 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     8101 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     8161 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     8221 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     8281 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     8341 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     8401 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8461 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8521 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8581 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8641 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8701 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8761 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8821 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8881 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     8941 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     9001 tcgtc
//
