LOCUS       sequence-224300-        6579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  studying clustering of M3 receptors by superresolution microscopy 
            (PALM).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6579)
  AUTHORS   Scarselli M, Annibale P, Radenovic A
  TITLE     Cell type-specific beta2-adrenergic receptor clusters identified 
            using photoactivated localization microscopy are not lipid raft 
            related, but depend on actin cytoskeleton integrity.
  JOURNAL   J Biol Chem. 2012 May 11;287(20):16768-80. doi: 
            10.1074/jbc.M111.329912. Epub 2012 Mar 22.
  PUBMED    22442147
REFERENCE   2  (bases 1 to 6579)
  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: 
            "10.1074/jbc.M111.329912"; journalName: "J Biol Chem"; date: 
            "2012-05-11- 11"; volume: "287"; issue: "20"; pages: "16768-80"
COMMENT     Sequence Label: M3-PSCFP2
FEATURES             Location/Qualifiers
     source          1..6579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        98..401
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        402..605
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     556..576
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             687..767
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             2565..3281
                     /codon_start=1
                     /product="photoswitchable cyan-to-green fluorescent 
                     protein"
                     /label=PS-CFP2
                     /note="mammalian codon-optimized"
                     /translation="MSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVATLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG
                     NYKTRAEVKFEGDTLVSRIELTGTDFKEDGNILGNKMEYNYNATNVYIVADKARNGIKV
                     NFKVRHNIKDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYLE
                     FVTAAAITHGMDELYK"
     polyA_signal    3404..3525
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3441..3460)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3495..3514
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3532..3987)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3669..3690)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3881..3900
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4014..4118
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4120..4477
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4120..4311
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4143..4163)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4328..4463
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4512..5306
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4512..5306
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4566..4585)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5176..5195
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5494..5513)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5538..5585
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5914..6502
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6403..6422
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
       61 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      121 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      181 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      241 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      301 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      361 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      421 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      481 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      541 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      601 gagctggttt agtgaaccgt cagatccgct agcgctaccg gactcagatc tcgagctcaa
      661 gcttcgaatt ccttggtacc accatgtacc catacgatgt tccagattac gcttacccat
      721 acgatgttcc agattacgct tacccatacg atgttccaga ttacgctgat accttgcaca
      781 ataacagtac aacctcgcct ttgtttccaa acatcagctc ctcctggata cacagcccct
      841 ccgatgcagg gctgcccccg ggaaccgtca ctcatttcgg cagctacaat gtttctcgag
      901 cagctggcaa tttctcctct ccagacggta ccaccgatga ccctctggga ggtcataccg
      961 tctggcaagt ggtcttcatc gctttcttaa cgggcatcct ggccttggtg accatcatcg
     1021 gcaacatcct ggtaattgtg tcatttaagg tcaacaagca gctgaagacg gtcaacaact
     1081 acttcctctt aagcctggcc tgtgccgatc tgattatcgg ggtcatttca atgaatctgt
     1141 ttacgaccta catcatcatg aatcgatggg ccttagggaa cttggcctgt gacctctggc
     1201 ttgccattga ctacgtagcc agcaatgcct ctgttatgaa tcttctggtc atcagctttg
     1261 acagatactt ttccatcacg aggccgctca cgtaccgagc caaacgaaca acaaagagag
     1321 ccggtgtgat gatcggtctg gcttgggtca tctcctttgt cctttgggct cctgccatct
     1381 tgttctggca atactttgtt ggaaagagaa ctgtgcctcc gggagagtgc ttcattcagt
     1441 tcctcagtga gcccaccatt acttttggca cagccatcgc tgctttttat atgcctgtca
     1501 ccattatgac tattttatac tggaggatct ataaggaaac tgaaaagcgt accaaagagc
     1561 ttgctggcct gcaagcctct gggacagagg cagagacaga aaactttgtc caccccacgg
     1621 gcagttctcg aagctgcagc agttacgaac ttcaacagca aagcatgaaa cgctccaaca
     1681 ggaggaagta tggccgctgc cacttctggt tcacaaccaa gagctggaaa cccagctccg
     1741 agcagatgga ccaagaccac agcagcagtg acagttggaa caacaatgat gctgctgcct
     1801 ccctggagaa ctccgcctcc tccgacgagg aggacattgg ctccgagacg agagccatct
     1861 actccatcgt gctcaagctt ccgggtcaca gcaccatcct caactccacc aagttaccct
     1921 catcggacaa cctgcaggtg cctgaggagg agctggggat ggtggacttg gagaggaaag
     1981 ccgacaagct gcaggcccag aagagcgtgg acgatggagg cagttttcca aaaagcttct
     2041 ccaagcttcc catccagcta gagtcagccg tggacacagc taagacttct gacgtcaact
     2101 cctcagtggg taagagcacg gccactctac ctctgtcctt caaggaagcc actctggcca
     2161 agaggtttgc tctgaagacc agaagtcaga tcactaagcg gaaaaggatg tccctggtca
     2221 aggagaagaa agcggcccag accctcagtg cgatcttgct tgccttcatc atcacttgga
     2281 ccccatacaa catcatggtt ctggtgaaca ccttttgtga cagctgcata cccaaaacct
     2341 tttggaatct gggctactgg ctgtgctaca tcaacagcac cgtgaacccc gtgtgctatg
     2401 ctctgtgcaa caaaacattc agaaccactt tcaagatgct gctgctgtgc cagtgtgaca
     2461 aaaaaaagag gcgcaagcag cagtaccagc agagacagtc ggtcattttt cacaagcgcg
     2521 cacccgagca ggccttgggg gcccgggatc caccggtcgc caccatgagc aagggcgccg
     2581 agctgttcac cggcatcgtg cccatcctga tcgagctgaa tggcgatgtg aatggccaca
     2641 agttcagcgt gagcggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     2701 tcatctgcac caccggcaag ctgcctgtgc cctggcccac cctggtggcc accctgagct
     2761 acggcgtgca gtgcttctca cgctaccccg atcacatgaa gcagcacgac ttcttcaaga
     2821 gcgccatgcc tgagggctac atccaggagc gcaccatctt cttcgaggat gacggcaact
     2881 acaagacgcg cgccgaggtg aagttcgagg gcgataccct ggtgagtcgc atcgagctga
     2941 ccggcactga tttcaaggag gatggcaaca tcctgggcaa taagatggag tacaactaca
     3001 acgccaccaa tgtgtacatc gtggccgaca aggccaggaa tggcatcaag gtgaacttca
     3061 aggtccgcca caacatcaag gatggcagcg tgcagctggc cgaccactac cagcagaata
     3121 cccccatcgg cgatggccct gtgctgctgc ccgataacca ctacctgtcc acccagagcg
     3181 ccctgtccaa ggaccccaac gagaagcgcg atcacatgat ctacctcgag ttcgtgaccg
     3241 ccgccgccat cacccacggc atggatgagc tgtacaagta aagcggccgc gactctagat
     3301 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     3361 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3421 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3481 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     3541 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3601 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3661 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3721 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3781 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3841 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     3901 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     3961 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4021 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4081 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     4141 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4201 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4261 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4321 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4381 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4441 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     4501 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4561 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4621 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4681 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4741 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4801 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4861 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4921 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4981 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     5041 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5101 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5161 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5221 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5281 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5341 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5401 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5461 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     5521 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     5581 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     5641 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     5701 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     5761 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5821 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5881 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5941 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6001 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6061 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6121 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6181 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6241 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6301 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6361 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6421 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6481 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6541 ttgctggcct tttgctcaca tgttctttcc tgcgttatc
//
