LOCUS       sequence-224304-        6572 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 6572)
  TITLE     Radenovic Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6572)
  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            "Unpublished"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: M3-PAmcherry1
FEATURES             Location/Qualifiers
     source          1..6572
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        100..403
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        404..607
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     558..578
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             689..769
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     regulatory      2561..2570
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2567..3277
                     /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(2712..2730)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    3399..3520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3436..3455)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3490..3509
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3527..3982)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3664..3685)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3876..3895
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4009..4113
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4115..4472
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4115..4306
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4138..4158)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4323..4458
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4507..5301
                     /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             4507..5301
                     /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(4561..4580)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5171..5190
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5489..5508)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5533..5580
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5909..6497
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6398..6417
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat
       61 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      121 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      181 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      241 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      301 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      361 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      421 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      481 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      541 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      601 cagagctggt ttagtgaacc gtcagatccg ctagcgctac cggactcaga tctcgagctc
      661 aagcttcgaa ttccttggta ccaccatgta cccatacgat gttccagatt acgcttaccc
      721 atacgatgtt ccagattacg cttacccata cgatgttcca gattacgctg ataccttgca
      781 caataacagt acaacctcgc ctttgtttcc aaacatcagc tcctcctgga tacacagccc
      841 ctccgatgca gggctgcccc cgggaaccgt cactcatttc ggcagctaca atgtttctcg
      901 agcagctggc aatttctcct ctccagacgg taccaccgat gaccctctgg gaggtcatac
      961 cgtctggcaa gtggtcttca tcgctttctt aacgggcatc ctggccttgg tgaccatcat
     1021 cggcaacatc ctggtaattg tgtcatttaa ggtcaacaag cagctgaaga cggtcaacaa
     1081 ctacttcctc ttaagcctgg cctgtgccga tctgattatc ggggtcattt caatgaatct
     1141 gtttacgacc tacatcatca tgaatcgatg ggccttaggg aacttggcct gtgacctctg
     1201 gcttgccatt gactacgtag ccagcaatgc ctctgttatg aatcttctgg tcatcagctt
     1261 tgacagatac ttttccatca cgaggccgct cacgtaccga gccaaacgaa caacaaagag
     1321 agccggtgtg atgatcggtc tggcttgggt catctccttt gtcctttggg ctcctgccat
     1381 cttgttctgg caatactttg ttggaaagag aactgtgcct ccgggagagt gcttcattca
     1441 gttcctcagt gagcccacca ttacttttgg cacagccatc gctgcttttt atatgcctgt
     1501 caccattatg actattttat actggaggat ctataaggaa actgaaaagc gtaccaaaga
     1561 gcttgctggc ctgcaagcct ctgggacaga ggcagagaca gaaaactttg tccaccccac
     1621 gggcagttct cgaagctgca gcagttacga acttcaacag caaagcatga aacgctccaa
     1681 caggaggaag tatggccgct gccacttctg gttcacaacc aagagctgga aacccagctc
     1741 cgagcagatg gaccaagacc acagcagcag tgacagttgg aacaacaatg atgctgctgc
     1801 ctccctggag aactccgcct cctccgacga ggaggacatt ggctccgaga cgagagccat
     1861 ctactccatc gtgctcaagc ttccgggtca cagcaccatc ctcaactcca ccaagttacc
     1921 ctcatcggac aacctgcagg tgcctgagga ggagctgggg atggtggact tggagaggaa
     1981 agccgacaag ctgcaggccc agaagagcgt ggacgatgga ggcagttttc caaaaagctt
     2041 ctccaagctt cccatccagc tagagtcagc cgtggacaca gctaagactt ctgacgtcaa
     2101 ctcctcagtg ggtaagagca cggccactct acctctgtcc ttcaaggaag ccactctggc
     2161 caagaggttt gctctgaaga ccagaagtca gatcactaag cggaaaagga tgtccctggt
     2221 caaggagaag aaagcggccc agaccctcag tgcgatcttg cttgccttca tcatcacttg
     2281 gaccccatac aacatcatgg ttctggtgaa caccttttgt gacagctgca tacccaaaac
     2341 cttttggaat ctgggctact ggctgtgcta catcaacagc accgtgaacc ccgtgtgcta
     2401 tgctctgtgc aacaaaacat tcagaaccac tttcaagatg ctgctgctgt gccagtgtga
     2461 caaaaaaaag aggcgcaagc agcagtacca gcagagacag tcggtcattt ttcacaagcg
     2521 cgcacccgag caggccttgg gggcccggga tccaccggtc gccaccatgg tgagcaaggg
     2581 cgaggaggat aacatggcca tcattaagga gttcatgcgc ttcaaggtgc acatggaggg
     2641 gtccgtgaac ggccacgtgt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     2701 cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttca cctgggacat
     2761 cctgagccct cagttcatgt acggctccaa tgcctacgtg aagcaccccg ccgacatccc
     2821 cgactacttt aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaaattcga
     2881 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     2941 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtgatgc agaagaagac
     3001 catgggctgg gaggccctct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     3061 ggtcaagccc agagtgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     3121 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaacc gcaagctgga
     3181 catcaccagc cacaacgagg actacaccat cgtggagcag tacgagagag ccgagggccg
     3241 ccactccacc ggcggcatgg acgagctgta caagtaagcg gccgcgactc tagatcataa
     3301 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     3361 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3421 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3481 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     3541 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3601 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3661 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3721 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3781 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3841 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3901 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3961 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4021 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4081 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     4141 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4201 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4261 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4321 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4381 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     4441 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     4501 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4561 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4621 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4681 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4741 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4801 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4861 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4921 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4981 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     5041 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5101 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5161 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5221 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5281 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5341 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5401 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5461 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     5521 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     5581 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     5641 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     5701 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     5761 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5821 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5881 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5941 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6001 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6061 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6121 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6181 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6241 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6301 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6361 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6421 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6481 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6541 ggccttttgc tcacatgttc tttcctgcgt ta
//
