LOCUS       sequence-443019-        6195 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HaloTags to outer surface in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6195)
  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 6195)
  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-Halo
FEATURES             Location/Qualifiers
     source          1..6195
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        56..435
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        436..639
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     590..610
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     684..703
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        684..702
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     783..842
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             846..872
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             900..1784
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     CDS             1791..1820
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1824..1970
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(1992..2009)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1998..2222
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2354..2942)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2434..2453)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3271..3318
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3343..3362
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3550..4344)
                     /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(3550..4344)
                     /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(3661..3680)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4271..4290
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(4443..4462)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        4541..4732
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4689..4709
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4792..5583,5584..5652))
                     /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     5415..5434
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5653..5757)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5784..6195,1..44)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5871..5890)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6081..6102
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttacgcgc gcgttgacat
       61 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      121 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      181 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      241 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      301 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      361 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      421 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      481 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      541 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      601 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
      661 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agcttggtac
      721 cgagctcgga tccactagta acggccgcca gtgtgctgga attcggcttg gggatatcca
      781 ccatggagac agacacactc ctgctatggg tactgctgct ctgggttcca ggttccactg
      841 gtgactatcc atatgatgtt ccagattatg ctggggccca gccggccaga tcaggatccg
      901 aaatcggtac tggctttcca ttcgaccccc attatgtgga agtcctgggc gagcgcatgc
      961 actacgtcga tgttggtccg cgcgatggca cccctgtgct gttcctgcac ggtaacccga
     1021 cctcctccta cgtgtggcgc aacatcatcc cgcatgttgc accgacccat cgctgcattg
     1081 ctccagacct gatcggtatg ggcaaatccg acaaaccaga cctgggttat ttcttcgacg
     1141 accacgtccg cttcatggat gccttcatcg aagccctggg tctggaagag gtcgtcctgg
     1201 tcattcacga ctggggctcc gctctgggtt tccactgggc caagcgcaat ccagagcgcg
     1261 tcaaaggtat tgcatttatg gagttcatcc gccctatccc gacctgggac gaatggccag
     1321 aatttgcccg cgagaccttc caggccttcc gcaccaccga cgtcggccgc aagctgatca
     1381 tcgatcagaa cgtttttatc gagggtacgc tgccgatggg tgtcgtccgc ccgctgactg
     1441 aagtcgagat ggaccattac cgcgagccgt tcctgaatcc tgttgaccgc gagccactgt
     1501 ggcgcttccc aaacgagctg ccaatcgccg gtgagccagc gaacatcgtc gcgctggtcg
     1561 aagaatacat ggactggctg caccagtccc ctgtcccgaa gctgctgttc tggggcaccc
     1621 caggcgttct gatcccaccg gccgaagccg ctcgcctggc caaaagcctg cctaactgca
     1681 aggctgtgga catcggcccg ggtctgaatc tgctgcaaga agacaacccg gacctgatcg
     1741 gcagcgagat cgcgcgctgg ctgtctactc tggagatttc cggtgtcgac gaacaaaaac
     1801 tcatctcaga agaggatctg aatgctgtgg gccaggacac gcaggaggtc atcgtggtgc
     1861 cacactcctt gccctttaag gtggtggtga tctcagccat cctggccctg gtggtgctca
     1921 ccatcatctc ccttatcatc ctcatcatgc tttggcagaa gaagccacgt taggcggccg
     1981 ctcgagatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2041 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2101 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2161 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     2221 ggcttctgag gcggaaagaa ccagtggcgg taatacggtt atccacagaa tcaggggata
     2281 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2341 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2401 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2461 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2521 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2581 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2641 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2701 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2761 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2821 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2881 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2941 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3001 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3061 aatgaagttt taaatcaatc taaagtatat atgagtaacc tgaggctatg gcagggcctg
     3121 ccgccccgac gttggctgcg agccctgggc cttcacccga acttgggggg tggggtgggg
     3181 aaaaggaaga aacgcgggcg tattggcccc aatggggtct cggtggggta tcgacagagt
     3241 gccagccctg ggaccgaacc ccgcgtttat gaacaaacga cccaacaccg tgcgttttat
     3301 tctgtctttt tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt
     3361 cagttagcct ccccctaggg tgggcgaaga actccagcat gagatccccg cgctggagga
     3421 tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg
     3481 tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca
     3541 gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg
     3601 agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc
     3661 aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca
     3721 gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc
     3781 atgggtcacg acgagatcct cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc
     3841 ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc
     3901 catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc
     3961 cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg
     4021 agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct
     4081 tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca
     4141 cgatagccgc gctgcctcgt cttgcagttc attcagggca ccggacaggt cggtcttgac
     4201 aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat
     4261 tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc
     4321 gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcgatctt
     4381 tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg
     4441 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
     4501 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
     4561 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
     4621 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     4681 ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc aggactcttc
     4741 ctttttcaat aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4801 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4861 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4921 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4981 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5041 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5101 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5161 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5221 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5281 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5341 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5401 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5461 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5521 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5581 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5641 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5701 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5761 atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5821 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5881 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5941 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     6001 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     6061 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     6121 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     6181 tgagctgatt taaca
//
