LOCUS       sequence-263734-        7659 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mCherry targeted to the plasma membrane with an 
            amphipathic helix of RitC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7659)
  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 7659)
  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: pAV0607
FEATURES             Location/Qualifiers
     source          1..7659
                     /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             4220..4921
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="SKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQ
                     TAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFED
                     GGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGE
                     IKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEG
                     RHSTGGMDELYK"
     primer_bind     complement(4359..4377)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4627..4646
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      5155..5342
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5210..5234)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(5399..5419)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5399..5417)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5438..5454)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5438..5454)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5451..5473)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5462..5478
                     /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(5486..5492,5493..5510,5511..5516))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5531..5552
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5669..5686)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5840..6428)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5920..5939)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6599..7390,7391..7459))
                     /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     7222..7241
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7460..7564)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7632..7650
                     /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 cgggcccgct
     3361 agcatgcgat ctacgataat gagacggtgt ttgggttgtt tttactgaat ttttctttta
     3421 ttttatgaca tcttaatttt tatgttaatg attattctta tacttttatc tatctttgat
     3481 taaaactaat attcattctt ttgatcacta attattttta tagtatacga ttcaacccaa
     3541 ctataataca ttgtatcaag agaaattttt aattagatgt aatttaatag ccatttgata
     3601 aaaaaaaatt accttttttt ttgctagtgt taaattattg atcataacaa tgtcgaatga
     3661 atactaataa aatagggaca cgcgagttgc aaatacttga aatacctttg aagatttgtc
     3721 tgtgaatttt gataagattt ttatgaccat ctcttttata ttcgatatcg tgaactttca
     3781 ttttcttttg ttttttgctt ttcaagttaa ataacgcgga aattatattt ccttatatgt
     3841 ttataagagc ttttaaagga tttgctatat cgcttacagt actacaaccg gttgctcaat
     3901 gttatccgtt tccgcggtct tcttccgtgc gcattctgcc gtgaagtgct aacgctgtgt
     3961 gtggctttat caaaaaagtg gctcaaatat agcaatttca tacatattta aacacccatt
     4021 atacttaaat gtatatttta aaatatataa gagattttca ctgtataact gtgtatctta
     4081 tctatatgta atcaacggct tcataccacc tcagccagcc gtgttataac ttaccgttta
     4141 ccaactacat tttttgtaac gaaccaaaaa accctcaaaa gacaagaccg aattcgatat
     4201 caagcttatg agatctgcta gcaagggcga ggaggataac atggccatca tcaaggagtt
     4261 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     4321 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     4381 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     4441 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     4501 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     4561 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     4621 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     4681 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     4741 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     4801 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     4861 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     4921 ggtcgaccac aagaaaaagt caaagtgtcc cttttttgag acatctgctg cataccgcta
     4981 ctatattgat gatgttttcc atgcccttgt acgggagata cgtaggaaag aaaaggaggc
     5041 agtactggcc atggagaaaa aatctaagcc caaaaacagt gtatggaaga ggctaaaatc
     5101 accattccgg aagaagaaag attcagtaac ttgagcggcc gcacttctaa ataagcgaat
     5161 ttcttatgat ttatgatttt tattattaaa taagttataa aaaaaataag tgtatacaaa
     5221 ttttaaagtg actcttaggt tttaaaacga aaattcttat tcttgagtaa ctctttcctg
     5281 taggtcaggt tgctttctca ggtatagtat gaggtcgctc ttattgacca cacctctacc
     5341 ggcagatccg ctagggataa cagggtaata tgcatgcaga gagctccagc ttttgttccc
     5401 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5461 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5521 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5581 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5641 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5701 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5761 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5821 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5881 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5941 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6001 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6061 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6121 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6181 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6241 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6301 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6361 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6421 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6481 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6541 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6601 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6661 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6721 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6781 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6841 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6901 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6961 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7021 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7081 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7141 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7201 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7261 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7321 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7381 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7441 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7501 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7561 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     7621 taacctataa aaataggcgt atcacgaggc cctttcgtc
//
