LOCUS       sequence-443299-        7383 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of CB1 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 7383)
  TITLE     Frank Lab Plasmids 2024
REFERENCE   2  (bases 1 to 7383)
  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"
COMMENT     Sequence Label: CMV-SNAP-hCB1_IRES_EGFP
FEATURES             Location/Qualifiers
     source          1..7383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(126..134,135..137,138..712)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(299..318)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     528..545
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(2178..2720)
                     /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"
     CDS             complement(2733..2759)
                     /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             complement(2733..2759)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(2887..3090)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2916..2936)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3091..3394
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3569..4157)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3649..3668)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4486..4533
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4558..4577
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4765..5559)
                     /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             complement(4765..5559)
                     /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(4876..4895)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5486..5505
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5594..5951)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5608..5743
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5760..5951
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5908..5928
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5953..6057)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6084..6539
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6171..6190)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6381..6402
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6546..6667
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6557..6576)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6611..6630
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6789..7383,1..125))
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
ORIGIN
        1 ttgccgtagg tggcatcgcc ctcgccctcg ccgctcacgc tgaacttgtg gccattcaca
       61 tcgccattca gctcgatcag gatgggcacg atgccggtga acagctcggc gcccttgctc
      121 accatggttg tggccatatt atcatcgtgt ttttcaaagg aaaaccacgt ccccgtggtt
      181 cggggggcct agacgttttt ttaacctcga ctaaacacat gtaaagcatg tgtaccgagg
      241 ccccagatca gatcccatac aatggggtac cttctgggca tccttcagcc ccttgttgaa
      301 tacgcttgag gagagccatt tgactctttc cacaactatc caactcacaa cgtggcactg
      361 gggttgtgcc gcctttgcag gtgtatctta tacacgtggc ttttggccgc agaggcacct
      421 gtcgccaggt ggggggttcc gctgcctgca aagggtcgct acagacgttg tttgtcttca
      481 agaagcttcc agaggaactg cttccttcac gacattcaac agaccttgca ttcctttggc
      541 gagaggggaa agacccctag gaatgctcgt caagaagaca gggccaggtt tccgggccct
      601 cacattgcca aaagacggca atatggtgga aaataacata tagacaaacg cacaccggcc
      661 ttattccaag cggcttcggc cagtaacgtt aggggggggg gagggagagg ggcggatccc
      721 gggcccgcgg taccgtcgac tgcagaattc gaagcttaca gagcctcggc agacgtgtct
      781 gtggacacag acatggttac cttggcaatc ttgaccgtgc tcttgatgca gctttctgcg
      841 gccctgtgaa cactggctgc attgtttgcg tgtttgtgca ggcagtccga gtcccccatg
      901 ctgttatcca gaggctgcgc agtgccttca caagagggaa acatgctccg gaaagcgtgt
      961 cgcaggtcct tactcctcag agcatagatg atggggttca cggtggagtt cagcaggcag
     1021 agcatactgc agaatgcaaa caccgtctta atgagcttgt tcatcttccc aaagacatca
     1081 tacaccatga ttgcaagcag agggccccag cagatgatca acaccaccag gatcaggacc
     1141 agggtcttgg ctaacctaat gtccatgcgg gcttggtctg gccgggtcac ctgtaccttc
     1201 ccatcctcag acgtgtggat gatgatgctc ttctgggtgc cacgctgaat catgcggacg
     1261 gcgtggctgt gagccttcca gagaatatac atgtacgcat acacgatgaa cagaagcagt
     1321 acgctggtga ccccgatcca gaacatcagg taggtttcat caatgtgtgg gaaaatgtct
     1381 gagcaaacag attgcagttt ctcgcagttc cagcccagga gaggcagcac ggcgatcaca
     1441 atggctatgg tccacatcag gcaaaacgcc accacggcct tgggcctggt gacaatcctc
     1501 ttataggcca ggggcctgtg aatggatatg tacctgtcga tggctgtgag gaacaggctg
     1561 cccacggagg cagtgaagga ggccgtgacc ccacccagtt tgaacagaaa cacgttgcgg
     1621 ctatctttgc ggtggaacac gtggaagtca atgaagctgt agacaaaaat gacactcccc
     1681 aggaggtctg ccaccgccag gctgccgatg aagtggtagg aaggcctgca gcggaggctg
     1741 cgggagtgga ggatgacgca cagcaccagg aggttctcca ggaccgtgaa ggtgcccagc
     1801 gtgagggaca ggactgcaat ggccagctgc tggctggggt tcaggaccat gaaacactct
     1861 atgtccatga agttctcccc acactggatg ttctcctcat tctccttgaa ggacgagaga
     1921 gacttgttgt aaaattctgt aatgttcacc tggtctgctg ggactagctg ggggttgtct
     1981 cccgcagtca tcttctcttg gaagggactt cccctaaagg aagttaaagg gaatttctgt
     2041 gggaagtacc ctaatttgga tgccatgtca cctttgatgt cttcgtactg aatgtcattt
     2101 gagcccacgt acaggaggtc agtggtgatg gtgcggaagg tggtatctgc aaggccatct
     2161 aggatcgact tacgcgtgcc cagcccaggc ttgcccagtc tgtggccctc gtgggccagc
     2221 agccactctt tcacggcgag cccgccctcg tagcccccca cggcgccaga gctagacacc
     2281 acccggtggc aggggatcag aatgggcacg ggatttccgc tcagggcggt tttcacggcg
     2341 gcggtggcgg cgggattgcc ggccagggcg gccagctgct ggtagctgat gacctctccg
     2401 aacttcacca ctttcagcag tttccacagc acctggcggg taaagctctc ctgctggaac
     2461 actgggtggt gcagggctgg cacagggaac tcctcgatgg cctcaggctg gtgaaagtag
     2521 gcgttgagcc aggcggtggc ctgcatcagt ggctctggtc cgcccagcac ggcggctggg
     2581 gcaggcactt ccacggcgtc ggcggcagat gttcctttgc ccagcagctt gatctcgtgc
     2641 aggccctgtt cgcacccaga cagttccagc ttgcccagag ggctatccag ggtggtgcgc
     2701 ttcatttcgc agtctttgtc ggatccacgc gtggcgtagt cgggcacgtc gtaggggtat
     2761 cgcgtggact gtgcactccc tcgtacatct tctttcagaa gtaggactga caggatcaac
     2821 agaaggacca tttgagctcg agatctgagt ccggtagcgc tagcggatct gacggttcac
     2881 taaaccagct ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
     2941 tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact
     3001 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
     3061 cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat
     3121 gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca
     3181 tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc
     3241 caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg
     3301 cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca
     3361 gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac
     3421 taatgacccc gtaattgatt actattaata actaatgcat ggcggtaata cggttatcca
     3481 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3541 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3601 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3661 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3721 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3781 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3841 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3901 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3961 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4021 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4081 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4141 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4201 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4261 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taacctgagg
     4321 ctatggcagg gcctgccgcc ccgacgttgg ctgcgagccc tgggccttca cccgaacttg
     4381 gggggtgggg tggggaaaag gaagaaacgc gggcgtattg gccccaatgg ggtctcggtg
     4441 gggtatcgac agagtgccag ccctgggacc gaaccccgcg tttatgaaca aacgacccaa
     4501 caccgtgcgt tttattctgt ctttttattg ccgtcatagc gcgggttcct tccggtattg
     4561 tctccttccg tgtttcagtt agcctccccc tagggtgggc gaagaactcc agcatgagat
     4621 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
     4681 catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg
     4741 tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
     4801 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
     4861 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
     4921 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
     4981 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc tcgccttgag
     5041 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
     5101 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
     5161 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     5221 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     5281 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     5341 cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc agttcattca gggcaccgga
     5401 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     5461 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     5521 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
     5581 ctcttgatcg atctttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga
     5641 atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat
     5701 ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg
     5761 ggactatggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
     5821 ctggggactt tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc
     5881 ctgctgggga gcctggggac tttccacacc ctaactgaca cacattccac agctggttct
     5941 ttccgcctca ggactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6001 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6061 atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     6121 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     6181 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     6241 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     6301 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     6361 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     6421 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     6481 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     6541 cgccttaaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     6601 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     6661 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     6721 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atctagagtc
     6781 gcggccgctc acttgtacag ctcatccatg ccgtgggtga tggcggcggc ggtcacgaag
     6841 ccgaagtaga tcatgtgatc gcgcttctcg ttggggtcct tggacagggc gctctgggtg
     6901 gacaggtagt ggttatcggg cagcagcaca gggccatcgc cgatgggggt attctgctgg
     6961 tagtggtcgg ccagctgcac gctgccatcc tcgatgttgt ggcggatctt gaagttcacc
     7021 ttgatgccat tcttggcctt gtcggtcatg atgtacacat tgtgggcgtt gtagttgtac
     7081 tccatcttat tgcccaggat gttgccatcc tccttgaaat cggtgccggt cagctcgatg
     7141 cgattcacca gggtatcgcc ctcgaacttc acctcggcgc gcgacttgta gttgccgtca
     7201 tcctcgaaga agatggtgcg ctcctggatg tagccctcag gcatggcgct cttgaagaag
     7261 tcgtgctgct tcatgtgatc ggggtagcgt gagaagcact gcacgccgta gctcagggtg
     7321 gtcaccaggg tgggccaggg cacaggcagc ttgccggtgg tgcagatgaa cttcagggtc
     7381 agc
//
