LOCUS       sequence-379660-        6965 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mXFPe-tagged IFNGR2 at the plasma membrane, e.g. for 
            single molecule fluorescence microscopy.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6965)
  AUTHORS   Sotolongo Bellon J, Birkholz O, Richter CP, Eull F, Kenneweg H, 
            Wilmes S, Rothbauer U, You C, Walter MR, Kurre R, Piehler J
  TITLE     Four-color single-molecule imaging with engineered tags resolves the
            molecular architecture of signaling complexes in the plasma 
            membrane.
  JOURNAL   Cell Rep Methods. 2022 Feb 4;2(2):100165. doi: 
            10.1016/j.crmeth.2022.100165. eCollection 2022 Feb 28.
  PUBMED    35474965
REFERENCE   2  (bases 1 to 6965)
  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: "Cell Rep 
            Methods."; date: "2022-02-4"; volume: "2(2):100165. doi"; pages: " 
            10.1016/j.crmeth.2022.100165. eCollection 2022 Feb 28"
COMMENT     Sequence Label: pSems-leader-mXFPe-IFNGR2(30-337)
FEATURES             Location/Qualifiers
     source          1..6965
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(177..196)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        368..747
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        748..951
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     902..922
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     996..1015
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        996..1014
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     1051..1110
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             1114..1140
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(1150..1152,1153..1155,1156..1866)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTFGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDDHFLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1195..1216)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1456..1475)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1803..1824
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          2874..3103
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3086..3105
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(3176..3750,3751..3753,3754..3762)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3343..3360)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3570..3589
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3785..4588
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3848..3867)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4458..4477
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4718..4839
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4755..4774)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4941..4957)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4941..4957)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4954..4976)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4965..4981
                     /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(4989..4995,4996..5013,5014..5019))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5034..5055
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5172..5189)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5343..5931)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5423..5442)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6102..6893,6894..6962))
                     /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     6725..6744
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6963..6965,1..102))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
       61 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
      121 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
      181 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
      241 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
      301 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
      361 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      421 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      481 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      541 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      601 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      661 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      721 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      781 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      841 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      901 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
      961 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
     1021 caagcttggt accgagctcg gatcgatatc atggagacag acacactcct gctatgggta
     1081 ctgctgctct gggttccagg ttccactggt gactatccat atgatgttcc agattatgct
     1141 gaattcacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1201 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1261 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1321 tggcccaccc tcgtgaccac cctgaccttt ggcgtgcagt gcttcagccg ctaccccgac
     1381 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1441 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1501 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1561 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     1621 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     1681 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1741 gacgatcact tcctgagcac ccagtccaaa ctgagcaaag accccaacga gaagcgcgat
     1801 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1861 tacaagggat ccctcgagct gcccgctcct cagcacccga agattcgcct gtacaacgca
     1921 gagcaggtcc tgagttggga gccagtggcc ctgagcaata gcacgaggcc tgttgtctac
     1981 caagtgcagt ttaaatacac cgacagtaaa tggttcacgg ccgacatcat gtccataggg
     2041 gtgaattgta cacagatcac agcaacagag tgtgacttca ctgccgccag tccctcagca
     2101 ggcttcccaa tggatttcaa tgtcactcta cgccttcgag ctgagctggg agcactccat
     2161 tctgcctggg tgacaatgcc ttggtttcaa cactatcgga atgtgactgt cgggcctcca
     2221 gaaaacattg aggtgacccc aggagaaggc tccctcatca tcaggttctc ctctcccttt
     2281 gacatcgctg atacctccac ggcctttttt tgttattatg tccattactg ggaaaaagga
     2341 ggaatccaac aggtcaaagg ccctttcaga agcaactcca tttcattgga taacttaaaa
     2401 ccctccagag tgtactgttt acaagtccag gcacaactgc tttggaacaa aagtaacatc
     2461 tttagagtcg ggcatttaag caacatatct tgctacgaaa caatggcaga tgcctccact
     2521 gagcttcagc aagtcatcct gatctccgtg ggaacatttt cgttgctgtc ggtgctggca
     2581 ggagcctgtt tcttcctggt cctgaaatat agaggcctga ttaaatactg gtttcacact
     2641 ccaccaagca tcccattaca gatagaagag tatttaaaag acccaactca gcccatctta
     2701 gaggccttgg acaaggacag ctcaccaaag gatgacgtct gggactctgt gtccattatc
     2761 tcgtttccgg aaaaggagca agaagatgtt ctccaaacgc tttgagcggc cgcatagata
     2821 actgatccag tgtgctggaa ttaattcgct gtctgcgagg gccagctgtt ggggtgagta
     2881 ctccctctca aaagcgggca tgacttctgc gctaagattg tcagtttcca aaaacgagga
     2941 ggatttgata ttcacctggc ccgcggtgat gcctttgagg gtggccgcgt ccatctggtc
     3001 agaaaagaca atctttttgt tgtcaagctt gaggtgtggc aggcttgaga tctggccata
     3061 cacttgagtg acaatgacat ccactttgcc tttctctcca caggtgtcca ctcccaggtc
     3121 caactgcagg tcgagcatgc atctagggcg gccaattccg cccctctccc ccccccccct
     3181 ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     3241 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     3301 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     3361 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     3421 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     3481 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     3541 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     3601 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     3661 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     3721 tggttttcct ttgaaaaaca cgatgataag cttgccacaa cccgggataa ttcctgcagc
     3781 caatatggga tcggccattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3841 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3901 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3961 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4021 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4081 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4141 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4201 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4261 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4321 gcgcatgccc gacggcgatg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4381 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4441 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4501 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4561 ctatcgcctt cttgacgagt tcttctgagg ggatcaattc tctagataac tgatcataat
     4621 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     4681 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     4741 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4801 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtcgagt gcattctagt
     4861 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     4921 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4981 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5041 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5101 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5161 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5221 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5281 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5341 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5401 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5461 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5521 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5581 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5641 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5701 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5761 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5821 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5881 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5941 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6001 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6061 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6121 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6181 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6241 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6301 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6361 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6421 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6481 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6541 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6601 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6661 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6721 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6781 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6841 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6901 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6961 atact
//
