LOCUS       sequence-181930-        8201 bp    DNA     circular SYN 14-APR-2025
DEFINITION  mammalian expression of TNFR1-tdEOS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8201)
  AUTHORS   Heidbreder M, Zander C, Malkusch S, Widera D, Kaltschmidt B, 
            Kaltschmidt C, Nair D, Choquet D, Sibarita JB, Heilemann M
  TITLE     TNF-alpha influences the lateral dynamics of TNF receptor I in 
            living cells.
  JOURNAL   Biochim Biophys Acta. 2012 Oct;1823(10):1984-9. doi: 
            10.1016/j.bbamcr.2012.06.026. Epub 2012 Jun 27.
  PUBMED    22749881
REFERENCE   2  (bases 1 to 8201)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.bbamcr.2012.06.026"; journalName: "Biochim Biophys Acta";
            date: "2012-10"; volume: "1823"; issue: "10"; pages: "1984-9"
COMMENT     Sequence Label: pTNFR1-tdEOS
FEATURES             Location/Qualifiers
     source          1..8201
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             904..927
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2347..3021
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="SAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVKE
                     GGPLPFAFDILTTAFHYGNRVFAEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIARN
                     DITMEGDTFYNKVRFHGVNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIRMALLLE
                     GNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPDN
                     ARR"
     CDS             3067..3744
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="SAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVKE
                     GGPLPFAFDILTTAFHYGNRVFAEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIARN
                     DITMEGDTFYNKVRFHGVNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIRMALLLE
                     GNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPDN
                     ARR"
     promoter        complement(3754..3772)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3755..3772)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3792..3809)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3798..4022
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4068..4496
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4155..4174)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4365..4386
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4505..4525)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4510..4839
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4690..4825
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4752..4771
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4906..5700
                     /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(4960..4979)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5570..5589
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5874..5995
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5911..5930)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5965..5984
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6044..6060)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6044..6060)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6057..6079)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6068..6084
                     /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(6092..6098,6099..6116,6117..6122))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6137..6158
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6275..6292)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6446..7034)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6526..6545)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7205..7996,7997..8065))
                     /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     7828..7847
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8066..8170)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 atggactaca aagacgatga cgataaagaa ttctgcagat atccatcaca ctggcggccg
      961 ctgtctggca tgggcctctc caccgtgcct gacctgctgc tgccactggt gctcctggag
     1021 ctgttggtgg gaatataccc ctcaggggtt attggactgg tccctcacct aggggacagg
     1081 gagaagagag atagtgtgtg tccccaagga aaatatatcc accctcaaaa taattcgatt
     1141 tgctgtacca agtgccacaa aggaacctac ttgtacaatg actgtccagg cccggggcag
     1201 gatacggact gcagggagtg tgagagcggc tccttcaccg cttcagaaaa ccacctcaga
     1261 cactgcctca gctgctccaa atgccgaaag gaaatgggtc aggtggagat ctcttcttgc
     1321 acagtggacc gggacaccgt gtgtggctgc aggaagaacc agtaccggca ttattggagt
     1381 gaaaaccttt tccagtgctt caattgcagc ctctgcctca atgggaccgt gcacctctcc
     1441 tgccaggaga aacagaacac cgtgtgcacc tgccatgcag gtttctttct aagagaaaac
     1501 gagtgtgtct cctgtagtaa ctgtaagaaa agcctggagt gcacgaagtt gtgcctaccc
     1561 cagattgaga atgttaaggg cactgaggac tcaggcacca cagtgctgtt gcccctggtc
     1621 attttctttg gtctttgcct tttatccctc ctcttcattg gtttaatgta tcgctaccaa
     1681 cggtggaagt ccaagctcta ctccattgtt tgtgggaaat cgacacctga aaaagagggg
     1741 gagcttgaag gaactactac taagcccctg gccccaaacc caagcttcag tcccactcca
     1801 ggcttcaccc ccaccctggg cttcagtccc gtgcccagtt ccaccttcac ctccagctcc
     1861 acctataccc ccggtgactg tcccaacttt gcggctcccc gcagagaggt ggcaccaccc
     1921 tatcaggggg ctgaccccat ccttgcgaca gccctcgcct ccgaccccat ccccaacccc
     1981 cttcagaagt gggaggacag cgcccacaag ccacagagcc tagacactga tgaccccgcg
     2041 acgctgtacg ccgtggtgga gaacgtgccc ccgttgcgct ggaaggaatt cgtgcggcgc
     2101 ctagggctga gcgaccacga gatcgatcgg ctggagctgc agaacgggcg ctgcctgcgc
     2161 gaggcgcaat acagcatgct ggcgacctgg aggcggcgca cgccgcggcg cgaggccacg
     2221 ctggagctgc tgggacgcgt gctccgcgac atggacctgc tgggctgcct ggaggacatc
     2281 gaggaggcgc tttgcggccc cgccgccctc ccgcccgcgc ccagtcttct cagatttggc
     2341 ctcgagagtg cgattaagcc agacatgaag atcaacctcc gtatggaagg caacgtaaac
     2401 gggcaccact ttgtgatcga cggagatggt acaggcaagc cttttgaggg aaaacagagt
     2461 atggatcttg aagtcaaaga gggcggacct ctgccttttg cctttgatat cctgaccact
     2521 gcattccatt acggcaacag ggtattcgcc gaatatccag accacataca agactatttt
     2581 aagcagtcgt ttcctaaggg gtattcgtgg gaacgaagct tgactttcga agacgggggc
     2641 atttgcattg ccagaaacga cataacaatg gaaggggaca ctttctataa taaagttcga
     2701 tttcacggtg taaactttcc cgccaatggt ccagttatgc agaagaagac gctgaaatgg
     2761 gagccctcca ctgagaaaat gtatgtgcgt gatggagtgc tgaccggtga tattcgcatg
     2821 gctttgttgc ttgaaggaaa tgcccattac cgatgtgact tcagaactac ttacaaagct
     2881 aaggagaagg gtgtcaagtt accaggctac cactttgtgg accactgcat tgagatttta
     2941 agccatgaca aagattacaa caaggttaag ctgtatgagc atgctgttgc tcattctgga
     3001 ttgcctgaca atgccagacg atctagaggt cacggtactg gttctactgg ttctggttct
     3061 tctgagagtg cgattaagcc agacatgaag atcaacctcc gtatggaagg caacgtaaac
     3121 gggcaccact ttgtgatcga cggagatggt acaggcaagc cttttgaggg aaaacagagt
     3181 atggatcttg aagtcaaaga gggcggacct ctgccttttg cctttgatat cctgaccact
     3241 gcattccatt acggcaacag ggtattcgcc gaatatccag accacataca agactatttt
     3301 aagcagtcgt ttcctaaggg gtattcgtgg gaacgaagct tgactttcga agacgggggc
     3361 atttgcattg ccagaaacga cataacaatg gaaggggaca ctttctataa taaagttcga
     3421 tttcacggtg taaactttcc cgccaatggt ccagttatgc agaagaagac gctgaaatgg
     3481 gagccctcca ctgagaaaat gtatgtgcgt gatggagtgc tgaccggtga tattcgcatg
     3541 gctttgttgc ttgaaggaaa tgcccattac cgatgtgact tcagaactac ttacaaagct
     3601 aaggagaagg gtgtcaagtt accaggctac cactttgtgg accactgcat tgagatttta
     3661 agccatgaca aagattacaa caaggttaag ctgtatgagc atgctgttgc tcattctgga
     3721 ttgcctgaca atgccagacg ataagggccc tattctatag tgtcacctaa atgctagagc
     3781 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3841 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3901 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3961 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     4021 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     4081 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4141 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4201 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4261 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4321 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4381 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4441 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4501 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4561 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4621 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4681 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4741 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4801 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4861 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4921 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4981 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5041 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     5101 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5161 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5221 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5281 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5341 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5401 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     5461 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5521 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5581 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5641 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5701 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5761 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5821 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5881 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5941 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6001 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     6061 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     6121 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     6181 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6241 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6301 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6361 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6421 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6481 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6541 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6601 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6661 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6721 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6781 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6841 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6901 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6961 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7021 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7081 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7141 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7201 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7261 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7321 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7381 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7441 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7501 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7561 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7621 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7681 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7741 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7801 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7861 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7921 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7981 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8041 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8101 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8161 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
