LOCUS       sequence-263725-        8316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mCherry targeted to the endoplasmic reticulum with a 
            signal sequence and a retention signal (AHDL).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8316)
  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 8316)
  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: pAV0764
FEATURES             Location/Qualifiers
     source          1..8316
                     /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(2526..2981)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2663..2684)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2875..2894
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3107..3129
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3121..3138
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3122..3138
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3148..3167
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3148..3166
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4266..4973
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4411..4429)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4679..4698
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        5055..5398
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      5797..5994
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(6056..6076)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6056..6074)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6095..6111)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6095..6111)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6108..6130)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6119..6135
                     /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(6143..6149,6150..6167,6168..6173))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6188..6209
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6326..6343)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6497..7085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6577..6596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7256..8047,8048..8116))
                     /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     7879..7898
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8117..8221)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8289..8307
                     /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 gtatactagt
      661 aaaaatcaag tgaattgtgc ctaggatagg tagctaacgt gactgccact gaacctgtgt
      721 agcgtattag atccatcaag atcccttctg ctcatatcat tgctgatctg taaattgacc
      781 gcatcaaatt tgatctcatt tatatcgaga ttgatctata ttattttgcc aatttgcagt
      841 atataaactt atttgattaa gtttttggtc tccttcgcca cgtcttcatt ttgttagttt
      901 aaattttttt gactaactta gaaagggaag tttttaaaaa cgttgaaata ctttttcgat
      961 ttaaaaatat atatttaatt aattttctgt tccaaaatgg ttgctcctca tctttggtta
     1021 cgtgctgaaa ccaaaccttt agaagagcgc agtgctttga ctcctagaac tgccaagata
     1081 ctcgccgatg ctggatttca aattaccatt gagcgctctt cccaacgtgc tttcaaggac
     1141 aaagaatttg agcgccttgg tttccccatg gttcctgagg gctcatggcg acatgcacct
     1201 aaggacgcgt acattattgg tctcaaagag cttccggaaa atgataacag tcctttgaag
     1261 catactcata ttcaatttgc tcattgctac aaaaaccagg aaggttggcg tgaagtcttg
     1321 tcccgattcc ccgctggcaa tggtcttcta tatgatttgg aatttttgca agatgacaat
     1381 ggcagacgtg ttgccgcttt tggttaccac gctggttttg ctggatcagc tatctcatgt
     1441 ttagtttggg cacatcaatt attgcatcct aacaaacaat tccctgctat ccgtccgttc
     1501 cccaacgaaa aatctttggt tcgccatgtt gctcgtcaag tacgtttagc tttgaaaaag
     1561 aataacaacc agtatcctcg tattctggtt attggggctc ttggtcgttg tggtactgga
     1621 gcgtgcgatt tagcatcaaa gattggtatt cccttcgata acattcttcg ctgggacatt
     1681 aatgaaacta agaagggtgg tccctttacc gaaattacgg aatcagacat atttgttaac
     1741 tgcatttatc tgagcatgcc tattcccaaa ttctgtaccg tcgagtcgtt aaatgtacct
     1801 aaccgtaaat tgcgtgttgt ttgtgatgtt agctgtgaca ctaccaatcc taacaatcct
     1861 attccaattt ataatgtaaa cactacattt gatcatccta cagttgaggt taaaggtgtc
     1921 acaactcctc ctcctttaga ggtcatttct attgatcatc ttcctacttt attgccaagg
     1981 gaatcttccg aagcatttag tgaagctcta attcctagtc ttttggcttt gaaggacgtc
     2041 gacaatgctc ccgtttgggt cagagctaaa aaactttatg aaaccatggt tcaaaagctt
     2101 taaaataaat gttatttaat ttcaaagttc cgggtccttt atattgtttc accaaattat
     2161 tgaatattat tttctcaaaa attaacttta gaatacaatt tagacggcat tttcgatgct
     2221 ctggacattt tttccacgta cttgttaaat tgtagcggtg ttgcataata ctaccttttg
     2281 atttgttatt cacgttacaa tatatatttg actaatttaa atatgataat atacacattt
     2341 ctctgcaaac ctcctttata cttttcgccg gtcattgatt aataattaag aaacaggatt
     2401 tcatcctgca tttgccactt acctatatgg taaatataaa tcttgaaact ttccccaata
     2461 tttattgctt gctagggcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     2521 tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     2581 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     2641 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     2701 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     2761 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     2821 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     2881 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     2941 caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg
     3001 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     3061 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     3121 tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc
     3181 gtgccaaggg aagggtaggt gtcgttaagg tctgaggttt ggtgtcagtg ggagatctaa
     3241 aatattatgg caaaattttc ggcgtcgcaa caataatagt aaacagatgt aaggactata
     3301 ccattttgag atgttaaaat ggtgagcaat gatacaagtg atgatctatc atcttttgct
     3361 gattgataaa aaaagacaaa ccctttaatt atttttagct acataatttt ctcttaatgg
     3421 attaatttgg aaagtgaacg caaaatatta tttagtctaa acatgatcgt atttgagatg
     3481 atgccttttg caagtatgtt atactttatt taaccatgac ttttacaatt caaccaatgc
     3541 gacactatcg gacagttgaa gccaatttta attattagct atatatttta gcgtttttcc
     3601 attatatttg tagttaaaac agctatttat aggccattat caaatatata tataattttt
     3661 tttttttttt tttttctctt tcatttggga atatagtgaa ataactctat ataacacaat
     3721 atcaaaataa agatttccca ccttcactgc ttttgttctt ttcacaatta ttcatttcag
     3781 aatttaccaa tttcgatttt cattgtttac attcattgct atgaaaacgt aaggtggtgg
     3841 cgcaatagga cttatcgaaa tgtacagaac tcactataga attgttgtgt tgatgagctt
     3901 caactgcatt cttctggaaa gtactagtat taacgacgtg actgctcctc tcgttactta
     3961 gctgatttct ggtacgctat taaactcatc caaaaccaac tattctagtt tggtaaatct
     4021 taatcaaaaa ctattaaaac ccgtttacta tttacttaac aggttgtttt caataattgg
     4081 gaattggctt gtgcctacga tctcttgtaa ttgaactaca catataagca tttataagtt
     4141 ggtaatcttc aaattcttgt ttattgaaaa tgaagaagtt ccagctattt agcattttaa
     4201 gctactttgt agctttattc ctcctaccta tggcttttgc tagtggatcc ggtggtcccg
     4261 gtggtatggt gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct
     4321 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
     4381 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
     4441 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
     4501 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
     4561 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
     4621 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
     4681 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
     4741 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
     4801 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     4861 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     4921 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aaggaggccg
     4981 atgaacttta agcggccgcc accgcggtgg agctcgttta gcttgcctcg tccccgccgg
     5041 gtcacccggc cagcgacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca
     5101 gctcaggggc atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat
     5161 catttgcatc catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc
     5221 tgcagacctg cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg
     5281 cgcccctgta gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac
     5341 ttccttttaa aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaaccat
     5401 gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat
     5461 ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt
     5521 cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg
     5581 cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa
     5641 caggggcatc ttgagcccct gcggacggtg tcgacaggtg cttctcgatc tgcatcctgg
     5701 gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt
     5761 gctgccctct ggttatgtgt gggagggcta atcagtactg acaataaaaa gattcttgtt
     5821 ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat
     5881 gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt
     5941 gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg
     6001 attcgatact aacgccgcca tccacctgca ggtaatcacg tgccagcttt tgttcccttt
     6061 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6121 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6181 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6241 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6301 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6361 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6421 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6481 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6541 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6601 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6661 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6721 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6781 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6841 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6901 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6961 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7021 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7081 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7141 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7201 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7261 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7321 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7381 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7441 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7501 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7561 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7621 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7681 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7741 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7801 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7861 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7921 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7981 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8041 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8101 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8161 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8221 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     8281 cctataaaaa taggcgtatc acgaggccct ttcgtc
//
