LOCUS       sequence-209973-        6306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of membrane localized mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6306)
  AUTHORS   Ariotti N, Rae J, Giles N, Martel N, Sierecki E, Gambin Y, Hall TE, 
            Parton RG
  TITLE     Ultrastructural localisation of protein interactions using 
            conditionally stable nanobodies.
  JOURNAL   PLoS Biol. 2018 Apr 5;16(4):e2005473. doi: 
            10.1371/journal.pbio.2005473. eCollection 2018 Apr.
  PUBMED    29621251
REFERENCE   2  (bases 1 to 6306)
  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: "PLoS 
            Biol."; date: "2018-04-5"; volume: "16(4):e2005473. doi"; pages: " 
            10.1371/journal.pbio.2005473. eCollection 2018 Apr"
COMMENT     Sequence Label: mCherry-CaaX(Hras)
FEATURES             Location/Qualifiers
     source          1..6306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(163..184)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(533..544,545..707)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     1080..1096
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1116..1136
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        1160..2144
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        2179..2197
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2179..2196
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    2197..2221
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2227..2934
                     /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(2372..2390)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2640..2659
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2834..2857
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(2999..3023)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3025..3159
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(3222..3242)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(3250..3266)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    3282..3416
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(3855..4042,4043..4054)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(4088..4104)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4088..4104)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4141..4161)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4141..4159)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4180..4196)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4193..4215)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4204..4220
                     /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(4228..4234,4235..4252,4253..4258))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4273..4294
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4411..4428)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4582..5170)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4662..4681)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5341..6132,6133..6201))
                     /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     5964..5983
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6202..6306)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
       61 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
      121 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
      181 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
      241 ggcccacgct ggctaagaac tcatcagcct ccccggtcca tctacccacg taccaatgca
      301 ccaattggcc acaatgacgg ctactacatg gtgccattcc ttcctcttta taggaatgga
      361 gactacctcc tgtccaacaa tgctcttgga tacgagtacg cctacctgtt ggacccaggt
      421 cattgcacaa caccagaaat gccctctgat ctgcaaaaga cgtgaatatc tgttcagaca
      481 cccatatcca ctctgttcca cacaggtcag aggtttgtcc aggagttctt gacagaggtg
      541 taaaaagtac tcaaaaattt tactcaagtg aaagtacaag tacttaggga aaattttact
      601 caattaaaag taaaagtatc tggctagaat cttacttgag taaaagtaaa aaagtactcc
      661 attaaaattg tacttgagta ttaaggaagt aaaagtaaaa gcaagaaaga aaactagaga
      721 ttcttgttta agcttttaat ctcaaaaaac attaaatgaa atgcatacaa ggttttatcc
      781 tgctttagaa ctgtttgtat ttaattatca aactataaga cagacaatct aatgccagta
      841 cacgctactc aaagttgtaa aacctcagat ttaacttcag tagaagctga ttctcaaaat
      901 tgttagtgtc aagcctagct cttttggggc tgaaaagcaa tcctgcagtg ctgaaaagcc
      961 tctcacaggc agccgatgcg ggaagaggtg tattagtctt gatagagagg ctgcaaatag
     1021 caggaaacgt gagcagagac tccctggtgt ctgaaacaca ggccagatgg gccctcgagc
     1081 aggaaacagc tatgaccatg attacgccaa gctatcaact ttgtatagaa aagttgaagg
     1141 cctcttcgct attacgccag tcgaccgcca attcaatatg gcgtatatgg actcatgcca
     1201 attcaatatg gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca
     1261 attcaatatg gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca
     1321 attcaatatg gcggacctgg cactgtgcca actggggagg ggtctacttg gcacggtgcc
     1381 aagtttgagg aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc
     1441 aataatggcg gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg
     1501 ccctatgcca atatggctat tggccaggtt caatactatg tattggccct atgccatata
     1561 gtattccata tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat
     1621 ataccatata tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt
     1681 ctctatatat ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc
     1741 ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca
     1801 ataggaccac ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc
     1861 acgcccccta ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat
     1921 taatagtaac ttggcaagta cattactatt ggaagtacgc cagggtacat tggcagtact
     1981 cccattgacg tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac
     2041 gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc
     2101 gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg gaaccgccat tctgcctggg
     2161 gacgtcggag caagcttgat ttaggtgaca ctatagacaa gtttgtacaa aaaagcaggc
     2221 tccaccatgg tgagcaaggg cgaggaggac aacatggcca tcatcaagga gttcatgcgc
     2281 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     2341 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
     2401 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     2461 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     2521 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     2581 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     2641 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     2701 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     2761 gacgccgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     2821 aacgtcaaca tcaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
     2881 tacgagcgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagaagctg
     2941 aaccctcctg atgagagtgg ccccggctgc atgagctgca agtgtgtgct ctcctgagac
     3001 ccagctttct tgtacaaagt gggggatcca gacatgataa gatacattga tgagtttgga
     3061 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     3121 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     3181 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt ccaactttat tatacatagt
     3241 tgataattca ctggccgtcg ttttacggta ccatcgatga tgatccagac atgataagat
     3301 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     3361 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     3421 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     3481 gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tcctctagat cagatctgcg
     3541 aagatacggc cacgggtgct cttgatcctg tggctgattt tggactgtgc tgctcgcagc
     3601 tgctgatgaa tcacatactt cctccatttt cttccactga ttgactgtta taatttccct
     3661 aatttccagg tcaaggtgct gtgcattgtg gtaatagatg tgacatgacg tcacttccaa
     3721 aggaccaatg aacatgtctg accaatttca tataatgtga aaacgatttt cataggcaga
     3781 ataaataaca tttaaattaa actgggcatc agcgcaattc aattggtttg gtaatagcaa
     3841 gggaaaatag aatgaagtga tctccaaaaa ataagtactt tttgactgta aataaaattg
     3901 taaggagtaa aaagtacttt tttttctaaa aaaatgtaat taagtaaaag taaaagtatt
     3961 gatttttaat tgtactcaag taaagtaaaa atccccaaaa ataatactta agtacagtaa
     4021 tcaagtaaaa ttactcaagt actttacacc tctggttctt gaccccctac cttcagcaag
     4081 cccagcagat ccactagttc tagagcggcc gccaccgcgg tggagctcca gcttttgttc
     4141 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4201 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4261 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4321 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4381 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4441 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4501 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4561 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4621 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4681 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4741 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4801 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4861 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4921 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4981 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5041 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5101 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5161 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5221 ctcacgttaa ggcattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5281 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5341 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5401 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5461 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5521 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5581 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5641 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5701 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5761 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5821 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5881 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5941 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6001 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6061 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6121 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6181 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6241 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6301 tccgcg
//
