LOCUS       Exported                6038 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  studying clustering of Beta2 receptors by superresolution microscopy
            (PALM).
ACCESSION   .
VERSION     .
KEYWORDS    ps-3HA-Beta2-PSCFP2-N
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6038)
  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 6038)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6038
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        96..399
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        400..603
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     554..574
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             675..755
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             2022..2738
                     /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    2861..2982
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2898..2917)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2952..2971
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2989..3444)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3126..3147)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3338..3357
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3471..3575
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3577..3934
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3600..3620)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3785..3920
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3847..3866
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3969..4763
                     /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(4023..4042)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4633..4652
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4951..4970)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4995..5042
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5371..5959
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5860..5879
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg
       61 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      121 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      181 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      241 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      301 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      361 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
      421 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      481 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
      541 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      601 gctggtttag tgaaccgtca gatccgctag cgctaccgga ctcagatctc gagctcaagc
      661 ttggtaccac catgtaccca tacgatgttc cagattacgc ttacccatac gatgttccag
      721 attacgctta cccatacgat gttccagatt acgctgatgg gcaacccggg aacggcagcg
      781 ccttcttgct ggcacccaat agaagccatg cgccggacca cgacgtcacg cagcaaaggg
      841 acgaggtgtg ggtggtgggc atgggcatcg tcatgtctct catcgtcctg gccatcgtgt
      901 ttggcaatgt gctggtcatc acagccattg ccaagttcga gcgtctgcag acggtcacca
      961 actacttcat cacttcactg gcctgtgctg atctggtcat gggcctggca gtggtgccct
     1021 ttggggccgc ccatattctt atgaaaatgt ggacttttgg caacttctgg tgcgagtttt
     1081 ggacttccat tgatgtgctg tgcgtcacgg ccagcattga gaccctgtgc gtgatcgcag
     1141 tggatcgcta ctttgccatt acttcacctt tcaagtacca gagcctgctg accaagaata
     1201 aggcccgggt gatcattctg atggtgtgga ttgtgtcagg ccttacctcc ttcttgccca
     1261 ttcagatgca ctggtaccgg gccacccacc aggaagccat caactgctat gccaatgaga
     1321 cctgctgtga cttcttcacg aaccaagcct atgccattgc ctcttccatc gtgtccttct
     1381 acgttcccct ggtgatcatg gtcttcgtct actccagggt ctttcaggag gccaaaaggc
     1441 agctccagaa gattgacaaa tctgagggcc gcttccatgt ccagaacctt agccaggtgg
     1501 agcaggatgg gcggacgggg catggactcc gcagatcttc caagttctgc ttgaaggagc
     1561 acaaagccct caagacgtta ggcatcatca tgggcacttt caccctctgc tggctgccct
     1621 tcttcatcgt taacattgtg catgtgatcc aggataacct catccgtaag gaagtttaca
     1681 tcctcctaaa ttggataggc tatgtcaatt ctggtttcaa tccccttatc tactgccgga
     1741 gcccagattt caggattgcc ttccaggagc ttctgtgcct gcgcaggtct tctttgaagg
     1801 cctatgggaa tggctactcc agcaacggca acacagggga gcagagtgga tatcacgtgg
     1861 aacaggagaa agaaaataaa ctgctgtgtg aagacctccc aggcacggaa gactttgtgg
     1921 gccatcaagg tactgtgcct agcgataaca ttgattcaca agggaggaat tgtagtacaa
     1981 atgactcact gctgggggcc cgggatccac cggtcgccac catgagcaag ggcgccgagc
     2041 tgttcaccgg catcgtgccc atcctgatcg agctgaatgg cgatgtgaat ggccacaagt
     2101 tcagcgtgag cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     2161 tctgcaccac cggcaagctg cctgtgccct ggcccaccct ggtggccacc ctgagctacg
     2221 gcgtgcagtg cttctcacgc taccccgatc acatgaagca gcacgacttc ttcaagagcg
     2281 ccatgcctga gggctacatc caggagcgca ccatcttctt cgaggatgac ggcaactaca
     2341 agacgcgcgc cgaggtgaag ttcgagggcg ataccctggt gagtcgcatc gagctgaccg
     2401 gcactgattt caaggaggat ggcaacatcc tgggcaataa gatggagtac aactacaacg
     2461 ccaccaatgt gtacatcgtg gccgacaagg ccaggaatgg catcaaggtg aacttcaagg
     2521 tccgccacaa catcaaggat ggcagcgtgc agctggccga ccactaccag cagaataccc
     2581 ccatcggcga tggccctgtg ctgctgcccg ataaccacta cctgtccacc cagagcgccc
     2641 tgtccaagga ccccaacgag aagcgcgatc acatgatcta cctcgagttc gtgaccgccg
     2701 ccgccatcac ccacggcatg gatgagctgt acaagtaaag cggccgcgac tctagatcat
     2761 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2821 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2881 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2941 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     3001 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3061 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3121 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3181 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3241 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3301 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3361 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3421 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3481 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3541 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     3601 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3661 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3721 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3781 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3841 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3901 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     3961 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4021 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4081 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4141 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4201 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4261 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4321 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4381 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4441 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     4501 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4561 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4621 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4681 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4741 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4801 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4861 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4921 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     4981 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     5041 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     5101 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     5161 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     5221 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5281 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5341 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5401 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5461 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5521 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5581 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5641 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5701 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5761 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5821 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5881 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5941 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6001 ctggcctttt gctcacatgt tctttcctgc gttatccc
//
