LOCUS       sequence-224309-        7451 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 7451)
  TITLE     Radenovic Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7451)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pcDNA3.1-Beta2-PAmcherry1
FEATURES             Location/Qualifiers
     source          1..7451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        103..482
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        483..686
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     637..657
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     731..750
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        731..749
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             772..795
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             817..897
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             2152..2862
                     /codon_start=1
                     /product="photoactivatable monomeric derivative of DsRed 
                     fluorescent protein (Subach et al., 2009)"
                     /label=PAmCherry1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHVFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFTWDILSPQFMYGSNAYVKHPADIPDYFKLSFPEGFKWERVMKF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEALSERMYPEDGALK
                     GEVKPRVKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNRKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2297..2315)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     promoter        complement(2872..2890)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2873..2890)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2910..2927)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2916..3140
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3186..3614
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3273..3292)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3483..3504
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3623..3643)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3628..3957
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3808..3943
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3870..3889
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4024..4818
                     /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(4078..4097)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4688..4707
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4992..5113
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5029..5048)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5083..5102
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5162..5178)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5162..5178)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5175..5197)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5186..5202
                     /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(5210..5216,5217..5234,5235..5240))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5255..5276
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5393..5410)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5564..6152)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5644..5663)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6323..7114,7115..7183))
                     /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     6946..6965
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7184..7288)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7363..7382)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
       61 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
      121 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      181 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      241 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      301 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      361 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      421 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      481 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      541 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      601 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      661 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc
      721 ttatcgaaat taatacgact cactataggg agacccaagc ttggtaccat ggactacaaa
      781 gacgatgacg ataaagaatt ccttggtacc accatgtacc catacgatgt tccagattac
      841 gcttacccat acgatgttcc agattacgct tacccatacg atgttccaga ttacgctgat
      901 gggcaacccg ggaacggcag cgccttcttg ctggcaccca atagaagcca tgcgccggac
      961 cacgacgtca cgcagcaaag ggacgaggtg tgggtggtgg gcatgggcat cgtcatgtct
     1021 ctcatcgtcc tggccatcgt gtttggcaat gtgctggtca tcacagccat tgccaagttc
     1081 gagcgtctgc agacggtcac caactacttc atcacttcac tggcctgtgc tgatctggtc
     1141 atgggcctgg cagtggtgcc ctttggggcc gcccatattc ttatgaaaat gtggactttt
     1201 ggcaacttct ggtgcgagtt ttggacttcc attgatgtgc tgtgcgtcac ggccagcatt
     1261 gagaccctgt gcgtgatcgc agtggatcgc tactttgcca ttacttcacc tttcaagtac
     1321 cagagcctgc tgaccaagaa taaggcccgg gtgatcattc tgatggtgtg gattgtgtca
     1381 ggccttacct ccttcttgcc cattcagatg cactggtacc gggccaccca ccaggaagcc
     1441 atcaactgct atgccaatga gacctgctgt gacttcttca cgaaccaagc ctatgccatt
     1501 gcctcttcca tcgtgtcctt ctacgttccc ctggtgatca tggtcttcgt ctactccagg
     1561 gtctttcagg aggccaaaag gcagctccag aagattgaca aatctgaggg ccgcttccat
     1621 gtccagaacc ttagccaggt ggagcaggat gggcggacgg ggcatggact ccgcagatct
     1681 tccaagttct gcttgaagga gcacaaagcc ctcaagacgt taggcatcat catgggcact
     1741 ttcaccctct gctggctgcc cttcttcatc gttaacattg tgcatgtgat ccaggataac
     1801 ctcatccgta aggaagttta catcctccta aattggatag gctatgtcaa ttctggtttc
     1861 aatcccctta tctactgccg gagcccagat ttcaggattg ccttccagga gcttctgtgc
     1921 ctgcgcaggt cttctttgaa ggcctatggg aatggctact ccagcaacgg caacacaggg
     1981 gagcagagtg gatatcacgt ggaacaggag aaagaaaata aactgctgtg tgaagacctc
     2041 ccaggcacgg aagactttgt gggccatcaa ggtactgtgc ctagcgataa cattgattca
     2101 caagggagga attgtagtac aaatgactca ctgctgaagc ggccgctcga gatggtgagc
     2161 aagggcgagg aggataacat ggccatcatt aaggagttca tgcgcttcaa ggtgcacatg
     2221 gaggggtccg tgaacggcca cgtgttcgag atcgagggcg agggcgaggg ccgcccctac
     2281 gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcacctgg
     2341 gacatcctga gccctcagtt catgtacggc tccaatgcct acgtgaagca ccccgccgac
     2401 atccccgact actttaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaaa
     2461 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
     2521 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt gatgcagaag
     2581 aagaccatgg gctgggaggc cctctccgag cggatgtacc ccgaggacgg cgccctgaag
     2641 ggcgaggtca agcccagagt gaagctgaag gacggcggcc actacgacgc tgaggtcaag
     2701 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caaccgcaag
     2761 ctggacatca ccagccacaa cgaggactac accatcgtgg agcagtacga gagagccgag
     2821 ggccgccact ccaccggcgg catggacgag ctgtacaagt aagggcccta ttctatagtg
     2881 tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     2941 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3001 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3061 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3121 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3181 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3241 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3301 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3361 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3421 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3481 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3541 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3601 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3661 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3721 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3781 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3841 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     3901 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     3961 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4021 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4081 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4141 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4201 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4261 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4321 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4381 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4441 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4501 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4561 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4621 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4681 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4741 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4801 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4861 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4921 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4981 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5041 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5101 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     5161 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5221 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5281 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5341 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5401 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5461 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5521 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5581 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5641 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5701 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5761 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5821 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5881 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5941 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6001 gaagaacagt atttggtatc tgcgctttgc tgaagccagt taccttcgga aaaagagttg
     6061 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6121 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6181 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6241 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6301 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6361 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6421 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6481 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6541 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6601 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6661 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6721 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6781 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6841 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6901 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6961 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7021 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7081 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7141 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7201 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7261 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     7321 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     7381 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     7441 gagcaaaatt t
//
