LOCUS       sequence-406232-        5832 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Display SNAPtag on the surface of mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5832)
  AUTHORS   Zhang W, Lu CH, Nakamoto ML, Tsai CT, Roy AR, Lee CE, Yang Y, Jahed 
            Z, Li X, Cui B
  TITLE     Curved adhesions mediate cell attachment to soft matrix fibres in 
            three dimensions.
  JOURNAL   Nat Cell Biol. 2023 Sep 28. doi: 10.1038/s41556-023-01238-1.
  PUBMED    37770566
REFERENCE   2  (bases 1 to 5832)
  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: "Nat Cell 
            Biol. 2023 Sep 28. doi: 10.1038/s41556-023-01238-1."
COMMENT     Sequence Label: pDisplay-SNAPtag
FEATURES             Location/Qualifiers
     source          1..5832
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        389..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     637..656
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        637..655
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     736..795
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             817..1362
                     /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="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     CDS             1381..1410
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1414..1560
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(1582..1599)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1588..1812
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(1944..2532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2024..2043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2861..2908
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2933..2952
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3140..3934)
                     /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
                     GLAPAELFARLKASKPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3251..3270)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3861..3880
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(4033..4052)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        4131..4322
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4279..4299
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4382..5173,5174..5242))
                     /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     5005..5024
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5243..5347)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5374..5829
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5461..5480)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5671..5692
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgcgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      421 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      481 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagaa ctctctggct
      601 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
      661 ccaagcttgg taccgaactc ggatccacta gtaacggccg ccagtgtgct ggaattcggc
      721 ttggggatat ccaccatgga gacagacaca ctcctgctat gggtactgct gctctgggtt
      781 ccaggttcca ctggtgacag atctgccgca ggcagcatgg acaaagactg cgaaatgaag
      841 cgcaccaccc tggatagccc tctgggcaag ctggaactgt ctgggtgcga acagggcctg
      901 cacgagatca agctgctggg caaaggaaca tctgccgccg acgccgtgga agtgcctgcc
      961 ccagccgccg tgctgggcgg accagagcca ctgatgcagg ccaccgcctg gctcaacgcc
     1021 tactttcacc agcctgaggc catcgaggag ttccctgtgc cagccctgca ccacccagtg
     1081 ttccagcagg agagctttac ccgccaggtg ctgtggaaac tgctgaaagt ggtgaagttc
     1141 ggagaggtca tcagctacca gcagctggcc gccctggccg gcaatcccgc cgccaccgcc
     1201 gccgtgaaaa ccgccctgag cggaaatccc gtgcccattc tgatcccctg ccaccgggtg
     1261 gtgtctagct ctggcgccgt ggggggctac gagggcgggc tcgccgtgaa agagtggctg
     1321 ctggcccacg agggccacag actgggcaag cctgggctgg gcagctcgct gcaggtcgac
     1381 gaacaaaaac tcatctcaga agaggatctg aatgctgtgg gccaggacac gcaggaggtc
     1441 atcgtggtgc cacactcctt gccctttaag gtggtggtga tctcagccat cctggccctg
     1501 gtggtgctca ccatcatctc ccttatcatc ctcatcatgc tttggcagaa gaagccacgt
     1561 taggcggccg ctcgagatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     1621 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     1681 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     1741 tggggcagga cagcaagggg gaggattggg aagacaatag caggcgtgct ggggatgcgg
     1801 tgggctctat ggcttctgag gcggaaagaa ccagtggcgg taatacggtt atccacagaa
     1861 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1921 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1981 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2041 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2101 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2161 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2221 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2281 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2341 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2401 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2461 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2521 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2581 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2641 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaacc tgaggctatg
     2701 gcagggcctg ccgccccgac gttggctgcg agccctgggc cttcacccga acttgggggg
     2761 tggggtgggg aaaaggaaga aacgcgggcg tattggcccc aatggggtct cggtggggta
     2821 tcgacagagt gccagccctg ggaccgaacc ccgcgtttat gaacaaacga cccaacaccg
     2881 tgcgttttat tctgtctttt tattgccgtc atagcgcggg ttccttccgg tattgtctcc
     2941 ttccgtgttt cagttagcct ccccctaggg tgggcgaaga actccagcat gagatccccg
     3001 cgctggagga tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag
     3061 aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt
     3121 tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct
     3181 gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa
     3241 gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca
     3301 gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc
     3361 aggcatcgcc atgggtcacg acgagatcct cgccgtcggg cttgctcgcc ttgagcctgg
     3421 cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa
     3481 gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg
     3541 ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt
     3601 tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca
     3661 gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg
     3721 tggccagcca cgatagccgc gctgcctcgt cttgcagttc attcagggca ccggacaggt
     3781 cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag
     3841 agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg
     3901 gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt
     3961 gatcgatctt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc
     4021 tcagaggccg ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag
     4081 aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg
     4141 ttgctgacta attgagatgc ttgctttgca tacttctgcc tgctggggag cctggggact
     4201 ttccacacct ggttgctgac taattgagat gcttgctttg catacttctg cctgctgggg
     4261 agcctgggga ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc
     4321 aggactcttc ctttttcaat aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4381 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4441 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4501 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4561 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4621 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4681 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4741 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4801 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4861 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4921 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4981 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5041 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5101 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5161 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5221 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5281 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5341 ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg
     5401 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     5461 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     5521 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     5581 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     5641 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     5701 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     5761 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     5821 ttacaattta cg
//
