LOCUS       sequence-224260-        7421 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  studying clustering of Beta2 receptors by superresolution microscopy
            (PALM).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7421)
  AUTHORS   Annibale P, Vanni S, Scarselli M, Rothlisberger U, Radenovic A
  TITLE     Identification of clustering artifacts in photoactivated 
            localization microscopy.
  JOURNAL   Nat Methods. 2011 Jun 12;8(7):527-8. doi: 10.1038/nmeth.1627.
  PUBMED    21666669
REFERENCE   2  (bases 1 to 7421)
  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: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2011-06-12- 12"; volume: "8"; 
            issue: "7"; pages: "527-8"
COMMENT     Sequence Label: pcDNA-Flag1-3HABeta2-mEos2
FEATURES             Location/Qualifiers
     source          1..7421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(16..35)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        207..586
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        587..790
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     741..761
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     835..854
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        835..853
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             876..899
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             921..1001
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             2256..2936
                     /codon_start=1
                     /product="monomeric variant of green-to-red photoswitchable
                     fluorescent protein EosFP (McKinney et al., 2009)"
                     /label=mEos2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIHMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     promoter        complement(2946..2964)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2947..2964)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2984..3001)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2990..3214
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3260..3688
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3347..3366)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3557..3578
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3697..3717)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3702..4031
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3882..4017
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3944..3963
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4098..4892
                     /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(4152..4171)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4762..4781
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5066..5187
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5103..5122)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5157..5176
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5236..5252)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5236..5252)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5249..5271)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5260..5276
                     /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(5284..5290,5291..5308,5309..5314))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5329..5350
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5467..5484)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5638..6226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5718..5737)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6397..7188,7189..7257))
                     /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     7020..7039
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7258..7362)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
       61 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
      121 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
      181 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
      241 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      301 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      361 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      421 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      481 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      541 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      601 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      661 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      721 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      781 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
      841 gactcactat agggagaccc aagcttggta ccatggacta caaagacgat gacgataaag
      901 aattccttgg taccaccatg tacccatacg atgttccaga ttacgcttac ccatacgatg
      961 ttccagatta cgcttaccca tacgatgttc cagattacgc tgatgggcaa cccgggaacg
     1021 gcagcgcctt cttgctggca cccaatagaa gccatgcgcc ggaccacgac gtcacgcagc
     1081 aaagggacga ggtgtgggtg gtgggcatgg gcatcgtcat gtctctcatc gtcctggcca
     1141 tcgtgtttgg caatgtgctg gtcatcacag ccattgccaa gttcgagcgt ctgcagacgg
     1201 tcaccaacta cttcatcact tcactggcct gtgctgatct ggtcatgggc ctggcagtgg
     1261 tgccctttgg ggccgcccat attcttatga aaatgtggac ttttggcaac ttctggtgcg
     1321 agttttggac ttccattgat gtgctgtgcg tcacggccag cattgagacc ctgtgcgtga
     1381 tcgcagtgga tcgctacttt gccattactt cacctttcaa gtaccagagc ctgctgacca
     1441 agaataaggc ccgggtgatc attctgatgg tgtggattgt gtcaggcctt acctccttct
     1501 tgcccattca gatgcactgg taccgggcca cccaccagga agccatcaac tgctatgcca
     1561 atgagacctg ctgtgacttc ttcacgaacc aagcctatgc cattgcctct tccatcgtgt
     1621 ccttctacgt tcccctggtg atcatggtct tcgtctactc cagggtcttt caggaggcca
     1681 aaaggcagct ccagaagatt gacaaatctg agggccgctt ccatgtccag aaccttagcc
     1741 aggtggagca ggatgggcgg acggggcatg gactccgcag atcttccaag ttctgcttga
     1801 aggagcacaa agccctcaag acgttaggca tcatcatggg cactttcacc ctctgctggc
     1861 tgcccttctt catcgttaac attgtgcatg tgatccagga taacctcatc cgtaaggaag
     1921 tttacatcct cctaaattgg ataggctatg tcaattctgg tttcaatccc cttatctact
     1981 gccggagccc agatttcagg attgccttcc aggagcttct gtgcctgcgc aggtcttctt
     2041 tgaaggccta tgggaatggc tactccagca acggcaacac aggggagcag agtggatatc
     2101 acgtggaaca ggagaaagaa aataaactgc tgtgtgaaga cctcccaggc acggaagact
     2161 ttgtgggcca tcaaggtact gtgcctagcg ataacattga ttcacaaggg aggaattgta
     2221 gtacaaatga ctcactgctg aagcggccgc tcgagatgag tgcgattaag ccagacatga
     2281 agatcaaact ccgtatggaa ggcaacgtaa acgggcacca ctttgtgatc gacggagatg
     2341 gtacaggcaa gccttttgag ggaaaacaga gtatggatct tgaagtcaaa gagggcggac
     2401 ctctgccttt tgcctttgat atcctgacca ctgcattcca ttacggcaac agggtattcg
     2461 ccaaatatcc agacaacata caagactatt ttaagcagtc gtttcctaag gggtattcgt
     2521 gggaacgaag cttgactttc gaagacgggg gcatttgcat tgccagaaac gacataacaa
     2581 tggaagggga cactttctat aataaagttc gattttatgg taccaacttt cccgccaatg
     2641 gtccagttat gcagaagaag acgctgaaat gggagccctc cactgagaaa atgtatgtgc
     2701 gtgatggagt gctgacgggt gatattcata tggctttgtt gcttgaagga aatgcccatt
     2761 accgatgtga cttcagaact acttacaaag ctaaggagaa gggtgtcaag ttaccaggct
     2821 accactttgt ggaccactgc attgagattt taagccatga caaagattac aacaaggtta
     2881 agctgtatga gcatgctgtt gctcattctg gattgcctga caatgccaga cgataagggc
     2941 cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct
     3001 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3061 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3121 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3181 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3241 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3301 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3361 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3421 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3481 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3541 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3601 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3661 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     3721 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3781 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3841 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3901 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3961 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4021 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     4081 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4141 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4201 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4261 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4321 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4381 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4441 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4501 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4561 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4621 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4681 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4741 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4801 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4861 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4921 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4981 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5041 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5101 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5161 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5221 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5281 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5341 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5401 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5461 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5521 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5581 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5641 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5701 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5761 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5821 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5881 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5941 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6001 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6061 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6121 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6181 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6241 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6301 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6361 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6421 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6481 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6541 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6601 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6661 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6721 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6781 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6841 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6901 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6961 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7021 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7081 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7141 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7201 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7261 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7321 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7381 gccacctgac gtcgacggat cgggagatct cccgatcccc t
//
