LOCUS       sequence-176167-        6300 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endoplasmic reticulum (ER)-targeted rsTagRFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6300)
  AUTHORS   Hertel F, Mo GC, Duwe S, Dedecker P, Zhang J
  TITLE     RefSOFI for Mapping Nanoscale Organization of Protein-Protein 
            Interactions in Living Cells.
  JOURNAL   Cell Rep. 2016 Jan 12;14(2):390-400. doi: 
            10.1016/j.celrep.2015.12.036. Epub 2015 Dec 31.
  PUBMED    26748717
REFERENCE   2  (bases 1 to 6300)
  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"; doi: 
            "10.1016/j.celrep.2015.12.036"; journalName: "Cell Rep"; date: 
            "2016-01-12- 12"; volume: "14"; issue: "2"; pages: "390-400"
COMMENT     Sequence Label: pcDNA3-ER-rsTagRFP
FEATURES             Location/Qualifiers
     source          1..6300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             88..783
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFWKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEAATEMLYPADGGLEGRGDMALK
                     LVGGGHLICNLKTTYRSKNPAKNLKMPGVYFVDHRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHK"
     promoter        complement(844..862)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(845..862)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(882..899)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    888..1112
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1158..1586
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1245..1264)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1455..1476
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1595..1615)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1685..2001
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1852..1987
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1914..1933
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2068..2862
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2122..2141)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2732..2751
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3036..3157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3073..3092)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3127..3146
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3206..3222)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3206..3222)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3219..3241)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3230..3246
                     /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(3254..3260,3261..3278,3279..3284))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3299..3320
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3437..3454)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3608..4196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3688..3707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4367..5158,5159..5227))
                     /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     4990..5009
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5228..5332)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5407..5426)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5598..5977
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5978..6181
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6132..6152
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6226..6245
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6226..6244
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 atggaccctg tggtggtgct ggggctctgt ctctcctgtt tgcttctcct ttcactctgg
       61 aaacagagct atgggggagg ggatccgatg agcgagctga ttaaggagaa catgcacatg
      121 aagctgtaca tggagggcac cgtgaacaac caccacttca agtgcacatc cgagggcgaa
      181 ggcaagccct acgagggcac ccagaccatg agaatcaagg tggtcgaggg cggccctctc
      241 cccttcgcct tcgacatcct ggctaccagc ttcatgtacg gcagccgcac cttcatcaac
      301 cacacccagg gcatccccga cttctggaag cagtccttcc ctgagggctt cacatgggag
      361 agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag
      421 gacggctgcc tcatctacaa cgtcaagctc agaggggtga acttcccatc caacggccct
      481 gtgatgcaga agaaaacact cggctgggag gccgccaccg agatgctgta ccccgctgac
      541 ggcggcctgg aaggcagagg ggacatggcc ctgaagctcg tgggcggggg ccacctgatc
      601 tgcaacttga agaccacata cagatccaag aatcccgcta agaacctcaa gatgcccggc
      661 gtctactttg tggaccacag actggaaaga atcaaggagg ccgacaaaga gacctacgtc
      721 gagcagcacg aggtggctgt ggccagatac tgcgacctcc ctagcaaact ggggcacaag
      781 taagaattct gcagatatcc atcacactgg cggccgctcg agcatgcatc tagagggccc
      841 tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag
      901 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
      961 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     1021 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     1081 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     1141 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     1201 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     1261 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     1321 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     1381 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     1441 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     1501 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     1561 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     1621 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     1681 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     1741 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     1801 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     1861 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     1921 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     1981 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     2041 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     2101 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     2161 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     2221 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     2281 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     2341 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     2401 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     2461 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     2521 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     2581 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     2641 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     2701 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     2761 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     2821 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     2881 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     2941 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     3001 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     3061 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3121 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     3181 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3241 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3301 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3361 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3421 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3481 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3541 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3601 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3661 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3721 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3781 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3841 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3901 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3961 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4021 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4081 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4141 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4201 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4261 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4321 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4381 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4441 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4501 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4561 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4621 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4681 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4741 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4801 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4861 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4921 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4981 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5041 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5101 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5161 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5221 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5281 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5341 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     5401 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     5461 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     5521 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
     5581 ggccagatat acgcgttgac attgattatt gactagttat taatagtaat caattacggg
     5641 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     5701 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     5761 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     5821 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     5881 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     5941 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     6001 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     6061 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     6121 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     6181 tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta
     6241 tagggagacc caagcttggt accccttgca gcttccagtt gtcaggaaga gaaggctccc
//
