LOCUS       sequence-379664-        7453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mXFPm-tagged IFNGR1 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 7453)
  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 7453)
  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-mXFPm-IFNGR1(18-489)
FEATURES             Location/Qualifiers
     source          1..7453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(172..191)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        363..742
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        743..946
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     897..917
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     991..1010
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        991..1009
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     1046..1105
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             1109..1135
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(1142..1144,1145..1147,1148..1858)
                     /codon_start=1
                     /product="enhanced CFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTFGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLKYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1187..1208)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1448..1467)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1795..1816
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          3357..3586
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3569..3588
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(3659..4233,4234..4236,4237..4245)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3826..3843)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4053..4072
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4268..5071
                     /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(4331..4350)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4941..4960
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5201..5322
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5238..5257)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5424..5440)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5424..5440)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5437..5459)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5448..5464
                     /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(5472..5478,5479..5496,5497..5502))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5517..5538
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5655..5672)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5826..6414)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5906..5925)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6585..7376,7377..7445))
                     /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     7208..7227
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7446..7453,1..97))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
       61 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
      121 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
      181 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
      241 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
      301 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
      361 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
      421 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      481 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      541 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      601 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      661 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      721 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      781 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      841 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      901 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag
      961 agaacccact gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc
     1021 ttggtaccga gctcggatcg atatcatgga gacagacaca ctcctgctat gggtactgct
     1081 gctctgggtt ccaggttcca ctggtgacta tccatatgat gttccagatt atgctgaatt
     1141 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1201 cggcgacgta aacggccaca ggttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1261 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     1321 cctcgtgacc accctgacct ttggcgtgca gtgcttcagc cgctaccccg accacatgaa
     1381 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gtaccatctt
     1441 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1501 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1561 caagctgaaa tacaactaca tcagccacaa cgtctatatc accgccgaca agcagaagaa
     1621 cggcatcaag gccaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     1681 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1741 ctacctgagc acccagtcca aactgagcaa agaccccaac gagaagcgcg atcacatggt
     1801 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagac
     1861 cggtgagatg ggcaccgcgg atctggggcc gtcctcagtg cctacaccaa ctaatgttac
     1921 aattgaatcc tataacatga accctatcgt atattgggag taccagatca tgccacaggt
     1981 ccctgttttt accgtagagg taaagaacta tggtgttaag aattcagaat ggattgatgc
     2041 ctgcatcaat atttctcatc attattgtaa tatttctgat catgttggtg atccatcaaa
     2101 ttctctttgg gtcagagtta aagccagggt tggacaaaaa gaatctgcct atgcaaagtc
     2161 agaagaattt gctgtatgcc gagatggaaa aattggacca cctaaactgg atatcagaaa
     2221 ggaggagaag caaatcatga ttgacatatt tcacccttca gtttttgtaa atggagacga
     2281 gcaggaagtc gattatgatc ccgaaactac ctgttacatt agggtgtaca atgtgtatgt
     2341 gagaatgaac ggaagtgaga tccagtataa aatactcacg cagaaggaag atgattgtga
     2401 cgagattcag tgccagttag cgattccagt atcctcactg aattctcagt actgtgtttc
     2461 agcagaagga gtcttacatg tgtggggtgt tacaactgaa aagtcaaaag aagtttgtat
     2521 taccattttc aatagcagta taaaaggttc tctttggatt ccagttgttg ctgctttact
     2581 actctttcta gtgcttagcc tggtattcat ctgtttttat attaagaaaa ttaatccatt
     2641 gaaggaaaaa agcataatat tacccaagtc cttgatctct gtggtaagaa gtgctacttt
     2701 agagacaaaa cctgaatcaa aatatgtatc actcatcacg tcataccagc cattttcctt
     2761 agaaaaggag gtggtctgtg aagagccgtt gtctccagca acagttccag gcatgcatac
     2821 cgaagacaat ccaggaaaag tggaacatac agaagaactt tctagtataa cagaagtggt
     2881 gactactgaa gaaaatattc ctgacgtggt cccgggcagc catctgactc caatagagag
     2941 agagagttct tcacctttaa gtagtaacca gtctgaacct ggcagcatcg ctttaaactc
     3001 gtatcactcc agaaattgtt ctgagagtga tcactccaga aatggttttg atactgattc
     3061 cagctgtctg gaatcacata gctccttatc tgactcagaa tttcccccaa ataataaagg
     3121 tgaaataaaa acagaaggac aagagctcat aaccgtaata aaagccccca cctcctttgg
     3181 ttatgataaa ccacatgtgc tagtggatct acttgtggat gatagcggta aagagtcctt
     3241 gattggttat agaccaacag aagattccaa agaattttca tgagatcagc ggccgcatag
     3301 ataactgatc cagtgtgctg gaattaattc gctgtctgcg agggccagct gttggggtga
     3361 gtactccctc tcaaaagcgg gcatgacttc tgcgctaaga ttgtcagttt ccaaaaacga
     3421 ggaggatttg atattcacct ggcccgcggt gatgcctttg agggtggccg cgtccatctg
     3481 gtcagaaaag acaatctttt tgttgtcaag cttgaggtgt ggcaggcttg agatctggcc
     3541 atacacttga gtgacaatga catccacttt gcctttctct ccacaggtgt ccactcccag
     3601 gtccaactgc aggtcgagca tgcatctagg gcggccaatt ccgcccctct cccccccccc
     3661 cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     3721 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     3781 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     3841 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     3901 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     3961 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     4021 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     4081 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca
     4141 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     4201 acgtggtttt cctttgaaaa acacgatgat aagcttgcca caacccggga taattcctgc
     4261 agccaatatg ggatcggcca ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4321 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4381 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4441 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4501 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4561 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4621 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4681 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4741 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4801 ggcgcgcatg cccgacggcg atgatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4861 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4921 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4981 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5041 cttctatcgc cttcttgacg agttcttctg aggggatcaa ttctctagat aactgatcat
     5101 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     5161 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     5221 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5281 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtcg agtgcattct
     5341 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     5401 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5461 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5521 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5581 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5641 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5701 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5761 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5821 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5881 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5941 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6001 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6061 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6121 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6181 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6241 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6301 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6361 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6421 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6481 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6541 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6601 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6661 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6721 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6781 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6841 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6901 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6961 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7021 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7081 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7141 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7201 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7261 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7321 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7381 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7441 ctcatactct tcc
//
