LOCUS       sequence-442861-        5877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of 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
FEATURES             Location/Qualifiers
     source          1..5877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(622..648)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     sig_peptide     complement(652..711)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     complement(791..810)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(792..810)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(855..1058)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(884..904)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1059..1438
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1450..1905)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1587..1608)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1799..1818
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1932..2036
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2037..2105,2106..2897)
                     /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     complement(2255..2274)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     enhancer        2957..3148
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2980..3000)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     3227..3246
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3345..4139
                     /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             3345..4139
                     /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(3399..3418)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4009..4028
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4327..4346)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4371..4418
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4747..5335
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5236..5255
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5467..5691
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5680..5697
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(5719..5865)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(join(5869..5877,1..21))
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
ORIGIN
        1 ttctgagatg agtttttgtt cgtcgacgcc ctggaacaga acttccaggc ggccgcccag
       61 gcctggttta cccagacggt gaccttcgtg cgccagcagc cattctttca cagccaggcc
      121 gccttcgtag ccacctacgt ccaggtcgcc ctgaaccaca cggtggcacg ggatcagaat
      181 tggcaccgga ttaccagaca gagcggtttt caccgccgca gttgccgctg ggttgccagc
      241 cagcgccgcc agatgggaat aggagatcac ctcgccgaat ttcactactt tcagcagttt
      301 ccacagtacc tgacgggtaa aggattcctg ctgaaatacc gggtgatgca gggccggtac
      361 agggaactct tcaatcgcct ctggctggtg aaaatatgcg ttcagccatg cggtagcctg
      421 catcagcggc tccgggccac ccagaaccgc agccggagcc ggtacttcga ccgcatccgc
      481 agcgctcgta cctttgccca ggaaaatgat ttcgtgcagg ccctgctcgc aaccgctcag
      541 ttccagttta cccagcgggc tgtccagggt agtgcgtttc atttcgcagt ctttatccat
      601 tgatctggcc ggctgggccc cagcataatc tggaacatca tatggatagt caccagtgga
      661 acctggaacc cagagcagca gtacccatag caggagtgtg tctgtctcca tggtggatat
      721 ccccaagccg aattccagca cactggcggc cgttactagt ggatccgagc tcggtaccaa
      781 gcttgggtct ccctatagtg agtcgtatta atttcgataa gccagtaagc agtgggttct
      841 ctagttagcc agagagctct gcttatatag acctcccacc gtacacgcct accgcccatt
      901 tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa
      961 aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc
     1021 tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat
     1081 acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag
     1141 gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga
     1201 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
     1261 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
     1321 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
     1381 ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcaa
     1441 cgcgcgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     1501 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     1561 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     1621 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     1681 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     1741 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     1801 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     1861 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     1921 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1981 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2041 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2101 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2161 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2221 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     2281 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2341 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2401 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2461 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2521 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2581 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2641 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2701 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     2761 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     2821 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     2881 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttat
     2941 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     3001 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3061 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3121 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3181 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3241 ccgaggcccg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3301 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3361 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3421 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3481 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     3541 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3601 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3661 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3721 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3781 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3841 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     3901 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     3961 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4021 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4081 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4141 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4201 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4261 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     4321 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     4381 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     4441 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     4501 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     4561 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     4621 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4681 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4741 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4801 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4861 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4921 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4981 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5041 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5101 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5161 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5221 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5281 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5341 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5401 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccactggtt ctttccgcct
     5461 cagaagccat agagcccacc gcatccccag catgcctgct attgtcttcc caatcctccc
     5521 ccttgctgtc ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg
     5581 atgcaatttc ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg
     5641 aaggcacggg ggaggggcaa acaacagatg gctggcaact agaaggcaca gtcgaggctg
     5701 atctcgagcg gccgcctaac gtggcttctt ctgccaaagc atgatgagga tgataaggga
     5761 gatgatggtg agcaccacca gggccaggat ggctgagatc accaccacct taaagggcaa
     5821 ggagtgtggc accacgatga cctcctgcgt gtcctggccc acagcattca gatcctc
//
