LOCUS       sequence-442862-        5877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of dead-mutant SNAP-tags to outer surface in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5877)
  AUTHORS   Tobias JM, Rajic G, Viray AEG, Icka-Araki D, Frank JA
  TITLE     Genetically-targeted photorelease of endocannabinoids enables 
            optical control of GPR55 in pancreatic beta-cells.
  JOURNAL   Chem Sci. 2021 Sep 15;12(40):13506-13512. doi: 10.1039/d1sc02527a. 
            eCollection 2021 Oct 20.
  PUBMED    34777770
REFERENCE   2  (bases 1 to 5877)
  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: 
            "10.1039/d1sc02527a"; journalName: "Chem Sci"; date: "2021-09-15- 
            15"; volume: "12"; issue: "40"; pages: "13506-13512"
COMMENT     Sequence Label: CMV-pDisplay-SNAP[C145A]
FEATURES             Location/Qualifiers
     source          1..5877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     45..66
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     enhancer        215..594
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        595..798
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     749..769
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     843..862
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        843..861
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     942..1001
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             1005..1031
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1632..1661
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1665..1811
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(1833..1850)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1839..2063
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2195..2783)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2275..2294)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3112..3159
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3184..3203
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3391..4185)
                     /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(3391..4185)
                     /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(3502..3521)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4112..4131
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(4284..4303)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        4382..4573
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4530..4550
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4633..5424,5425..5493))
                     /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     5256..5275
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5494..5598)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5625..5877,1..203)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5712..5731)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
       61 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
      121 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
      181 caaaatatta acgcttacaa tttacgcgcg cgttgacatt gattattgac tagttattaa
      241 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      301 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      361 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      421 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      481 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      541 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      601 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      661 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      721 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      781 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
      841 attaatacga ctcactatag ggagacccaa gcttggtacc gagctcggat ccactagtaa
      901 cggccgccag tgtgctggaa ttcggcttgg ggatatccac catggagaca gacacactcc
      961 tgctatgggt actgctgctc tgggttccag gttccactgg tgactatcca tatgatgttc
     1021 cagattatgc tggggcccag ccggccagat caatggataa agactgcgaa atgaaacgca
     1081 ctaccctgga cagcccgctg ggtaaactgg aactgagcgg ttgcgagcag ggcctgcacg
     1141 aaatcatttt cctgggcaaa ggtacgagcg ctgcggatgc ggtcgaagta ccggctccgg
     1201 ctgcggttct gggtggcccg gagccgctga tgcaggctac cgcatggctg aacgcatatt
     1261 ttcaccagcc agaggcgatt gaagagttcc ctgtaccggc cctgcatcac ccggtatttc
     1321 agcaggaatc ctttacccgt caggtactgt ggaaactgct gaaagtagtg aaattcggcg
     1381 aggtgatctc ctattcccat ctggcggcgc tggctggcaa cccagcggca actgcggcgg
     1441 tgaaaaccgc tctgtctggt aatccggtgc caattctgat cccggcacac cgtgtggttc
     1501 agggcgacct ggacgtaggt ggctacgaag gcggcctggc tgtgaaagaa tggctgctgg
     1561 cgcacgaagg tcaccgtctg ggtaaaccag gcctgggcgg ccgcctggaa gttctgttcc
     1621 agggcgtcga cgaacaaaaa ctcatctcag aagaggatct gaatgctgtg ggccaggaca
     1681 cgcaggaggt catcgtggtg ccacactcct tgccctttaa ggtggtggtg atctcagcca
     1741 tcctggccct ggtggtgctc accatcatct cccttatcat cctcatcatg ctttggcaga
     1801 agaagccacg ttaggcggcc gctcgagatc agcctcgact gtgccttcta gttgccagcc
     1861 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     1921 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     1981 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     2041 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagtggcg gtaatacggt
     2101 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2161 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2221 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2281 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2341 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2401 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2461 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2521 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2581 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2641 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2701 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2761 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2821 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     2881 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac
     2941 ctgaggctat ggcagggcct gccgccccga cgttggctgc gagccctggg ccttcacccg
     3001 aacttggggg gtggggtggg gaaaaggaag aaacgcgggc gtattggccc caatggggtc
     3061 tcggtggggt atcgacagag tgccagccct gggaccgaac cccgcgttta tgaacaaacg
     3121 acccaacacc gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg
     3181 gtattgtctc cttccgtgtt tcagttagcc tccccctagg gtgggcgaag aactccagca
     3241 tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca
     3301 acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt
     3361 ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa
     3421 ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca
     3481 ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc
     3541 cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat
     3601 attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc
     3661 cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc
     3721 ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg
     3781 gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat
     3841 gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc
     3901 gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg
     3961 aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttgcagtt cattcagggc
     4021 accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac
     4081 ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac
     4141 ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca
     4201 tcctgtctct tgatcgatct ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact
     4261 tctggaatag ctcagaggcc gggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
     4321 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
     4381 cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     4441 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct
     4501 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt
     4561 ctttccgcct caggactctt cctttttcaa taaatcaatc taaagtatat atgagtaaac
     4621 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4681 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4741 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4801 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4861 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4921 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4981 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5041 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5101 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5161 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5221 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5281 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5341 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5401 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5461 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5521 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5581 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg
     5641 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     5701 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     5761 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     5821 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgata
//
