LOCUS       sequence-443023-        7047 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of CB2 N-terminally fused SNAP-tag, with targeting 
            sequence to boost plasma membrane expression. Coupled with EGFP 
            transfection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7047)
  AUTHORS   Kosar M, Sykes DA, Viray AEG, Vitale RM, Sarott RC, Ganzoni RL, 
            Onion D, Tobias JM, Leippe P, Ullmer C, Zirwes EA, Guba W, Grether 
            U, Frank JA, Veprintsev DB, Carreira EM
  TITLE     Platform Reagents Enable Synthesis of Ligand-Directed Covalent 
            Probes: Study of Cannabinoid Receptor 2 in Live Cells.
  JOURNAL   J Am Chem Soc. 2023 Jul 19;145(28):15094-15108. doi: 
            10.1021/jacs.2c13629. Epub 2023 Jul 4.
  PUBMED    37401816
REFERENCE   2  (bases 1 to 7047)
  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: 
            "10.1021/jacs.2c13629"; journalName: "J Am Chem Soc"; date: 
            "2023-07-19- 19"; volume: "145"; issue: "28"; pages: "15094-15108"
COMMENT     Sequence Label: CMV-SNAP-hCB2_IRES_EGFP
FEATURES             Location/Qualifiers
     source          1..7047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      103..238
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             287..1081
                     /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             287..1081
                     /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(341..360)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     951..970
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1269..1288)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1313..1360
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1689..2277
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2178..2197
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2452..2755
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2756..2959
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2910..2930
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3087..3113
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             3087..3113
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3126..3668
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="optimized for mammalian expression"
                     /translation="DKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVEV
                     PAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLKV
                     VKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLAV
                     KEWLLAHEGHRLGKPGLG"
     misc_feature    join(4798..5372,5373..5375,5376..5384)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4965..4982)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5192..5211
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5385..6104
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    6226..6347
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6263..6282)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6317..6336
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6354..6809)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6491..6512)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6703..6722
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6836..6940
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6942..7047,1..252)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        join(6942..7047,1..86)
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6965..6985)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
       61 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
      121 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
      181 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
      241 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
      301 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
      361 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
      421 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
      481 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
      541 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
      601 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
      661 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
      721 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
      781 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
      841 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
      901 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
      961 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     1021 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     1081 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     1141 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     1201 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     1261 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     1321 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     1381 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     1441 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     1501 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     1561 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     1621 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1681 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1741 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1801 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     1861 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1921 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1981 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2041 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2101 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2161 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2221 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2281 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2341 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta
     2401 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     2461 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     2521 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     2581 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     2641 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     2701 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     2761 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     2821 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     2881 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     2941 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggactca
     3001 gatctcgagc tcaaatggtc cttctgttga tcctgtcagt cctacttctg aaagaagatg
     3061 tacgagggag tgcacagtcc acgcgatacc cctacgacgt gcccgactac gccacgcgtg
     3121 gatccgacaa agactgcgaa atgaagcgca ccaccctgga tagccctctg ggcaagctgg
     3181 aactgtctgg gtgcgaacag ggcctgcacg agatcaagct gctgggcaaa ggaacatctg
     3241 ccgccgacgc cgtggaagtg cctgccccag ccgccgtgct gggcggacca gagccactga
     3301 tgcaggccac cgcctggctc aacgcctact ttcaccagcc tgaggccatc gaggagttcc
     3361 ctgtgccagc cctgcaccac ccagtgttcc agcaggagag ctttacccgc caggtgctgt
     3421 ggaaactgct gaaagtggtg aagttcggag aggtcatcag ctaccagcag ctggccgccc
     3481 tggccggcaa tcccgccgcc accgccgccg tgaaaaccgc cctgagcgga aatcccgtgc
     3541 ccattctgat cccctgccac cgggtggtgt ctagctctgg cgccgtgggg ggctacgagg
     3601 gcgggctcgc cgtgaaagag tggctgctgg cccacgaggg ccacagactg ggcaagcctg
     3661 ggctgggcac gcgtgaggaa tgctgggtga cagagatagc caatggctcc aaggatggct
     3721 tggattccaa ccctatgaag gattacatga tcctgagtgg tccccagaag acagctgttg
     3781 ctgtgttgtg cactcttctg ggcctgctaa gtgccctgga gaacgtggct gtgctctatc
     3841 tgatcctgtc ctcccaccaa ctccgccgga agccctcata cctgttcatt ggcagcttgg
     3901 ctggggctga cttcctggcc agtgtggtct ttgcatgcag ctttgtgaat ttccatgttt
     3961 tccatggtgt ggattccaag gctgtcttcc tgctgaagat tggcagcgtg actatgacct
     4021 tcacagcctc tgtgggtagc ctcctgctga ccgccattga ccgatacctc tgcctgcgct
     4081 atccaccttc ctacaaagct ctgctcaccc gtggaagggc actggtgacc ctgggcatca
     4141 tgtgggtcct ctcagcacta gtctcctacc tgcccctcat gggatggact tgctgtccca
     4201 ggccctgctc tgagcttttc ccactgatcc ccaatgacta cctgctgagc tggctcctgt
     4261 tcatcgcctt cctcttttcc ggaatcatct acacctatgg gcatgttctc tggaaggccc
     4321 atcagcatgt ggccagcttg tctggccacc aggacaggca ggtgccagga atggcccgaa
     4381 tgaggctgga tgtgaggttg gccaagaccc tagggctagt gttggctgtg ctcctcatct
     4441 gttggttccc agtgctggcc ctcatggccc acagcctggc cactacgctc agtgaccagg
     4501 tcaagaaggc ctttgctttc tgctccatgc tgtgcctcat caactccatg gtcaaccctg
     4561 tcatctatgc tctacggagt ggagagatcc gctcctctgc ccatcactgc ctggctcact
     4621 ggaagaagtg tgtgaggggc cttgggtcag aggcaaaaga agaagccccg agatcctcag
     4681 tcaccgagac agaggctgat gggaaaatca ctccgtggcc agattccaga gatctagacc
     4741 tctctgattg ctaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccgccc
     4801 ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     4861 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     4921 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     4981 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     5041 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     5101 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     5161 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     5221 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtacac
     5281 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     5341 cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtg agcaagggcg
     5401 ccgagctgtt caccggcatc gtgcccatcc tgatcgagct gaatggcgat gtgaatggcc
     5461 acaagttcag cgtgagcggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     5521 agttcatctg caccaccggc aagctgcctg tgccctggcc caccctggtg accaccctga
     5581 gctacggcgt gcagtgcttc tcacgctacc ccgatcacat gaagcagcac gacttcttca
     5641 agagcgccat gcctgagggc tacatccagg agcgcaccat cttcttcgag gatgacggca
     5701 actacaagtc gcgcgccgag gtgaagttcg agggcgatac cctggtgaat cgcatcgagc
     5761 tgaccggcac cgatttcaag gaggatggca acatcctggg caataagatg gagtacaact
     5821 acaacgccca caatgtgtac atcatgaccg acaaggccaa gaatggcatc aaggtgaact
     5881 tcaagatccg ccacaacatc gaggatggca gcgtgcagct ggccgaccac taccagcaga
     5941 atacccccat cggcgatggc cctgtgctgc tgcccgataa ccactacctg tccacccaga
     6001 gcgccctgtc caaggacccc aacgagaagc gcgatcacat gatctacttc ggcttcgtga
     6061 ccgccgccgc catcacccac ggcatggatg agctgtacaa gtgagcggcc gcgactctag
     6121 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     6181 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     6241 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6301 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     6361 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     6421 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     6481 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     6541 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     6601 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     6661 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     6721 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     6781 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     6841 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     6901 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     6961 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     7021 atgcaaagca tgcatctcaa ttagtca
//
