LOCUS       sequence-450964-        7270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses fusion of HaloTag and TurboID in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7270)
  AUTHORS   Kwak C, Park C, Ko M, Im CY, Moon H, Park YH, Kim SY, Lee S, Kang 
            MG, Kwon HJ, Hong E, Seo JK, Rhee HW
  TITLE     Identification of proteomic landscape of drug-binding proteins in 
            live cells by proximity-dependent target ID.
  JOURNAL   Cell Chem Biol. 2022 Dec 15;29(12):1739-1753.e6. doi: 
            10.1016/j.chembiol.2022.10.001. Epub 2022 Oct 21.
  PUBMED    36272407
REFERENCE   2  (bases 1 to 7270)
  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: 
            "10.1016/j.chembiol.2022.10.001"; journalName: "Cell Chem Biol"; 
            date: "2022-12-15- 15"; volume: "29"; issue: "12"; pages: 
            "1739-1753.e6"
COMMENT     Sequence Label: pCDNA3 HaloTag-V5-TurboID
FEATURES             Location/Qualifiers
     source          1..7270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(493..597)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(672..691)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        863..1242
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1243..1446
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1397..1417
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1491..1510
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1491..1509
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1531..2421
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             2431..2472
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2473..3429
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     promoter        complement(3451..3469)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3452..3469)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3489..3506)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3495..3719
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3765..4193
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3852..3871)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4062..4083
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4202..4222)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4207..4536
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4387..4522
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4449..4468
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4603..5397
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4657..4676)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5267..5286
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5571..5692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5608..5627)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5662..5681
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5741..5757)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5741..5757)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5754..5776)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5765..5781
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5789..5795,5796..5813,5814..5819))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5834..5855
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5972..5989)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6143..6731)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6223..6242)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6902..7270,1..423,424..492))
                     /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"
ORIGIN
        1 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
       61 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
      121 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
      181 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
      241 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
      301 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
      361 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
      421 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
      481 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
      541 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
      601 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
      661 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
      721 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
      781 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
      841 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      901 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      961 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     1021 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     1081 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     1141 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     1201 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     1261 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     1321 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     1381 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     1441 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
     1501 cactataggg agacccaagc ttgcggccgc atggcagaaa tcggtactgg ctttccattc
     1561 gacccccatt atgtggaagt cctgggcgag cgcatgcact acgtcgatgt tggtccgcgc
     1621 gatggcaccc ctgtgctgtt cctgcacggt aacccgacct cctcctacgt gtggcgcaac
     1681 atcatcccgc atgttgcacc gacccatcgc tgcattgctc cagacctgat cggtatgggc
     1741 aaatccgaca aaccagacct gggttatttc ttcgacgacc acgtccgctt catggatgcc
     1801 ttcatcgaag ccctgggtct ggaagaggtc gtcctggtca ttcacgactg gggctccgct
     1861 ctgggtttcc actgggccaa gcgcaatcca gagcgcgtca aaggtattgc atttatggag
     1921 ttcatccgcc ctatcccgac ctgggacgaa tggccagaat ttgcccgcga gaccttccag
     1981 gccttccgca ccaccgacgt cggccgcaag ctgatcatcg atcagaacgt ttttatcgag
     2041 ggtacgctgc cgatgggtgt cgtccgcccg ctgactgaag tcgagatgga ccattaccgc
     2101 gagccgttcc tgaatcctgt tgaccgcgag ccactgtggc gcttcccaaa cgagctgcca
     2161 atcgccggtg agccagcgaa catcgtcgcg ctggtcgaag aatacatgga ctggctgcac
     2221 cagtcccctg tcccgaagct gctgttctgg ggcaccccag gcgttctgat cccaccggcc
     2281 gaagccgctc gcctggccaa aagcctgcct aactgcaagg ctgtggacat cggcccgggt
     2341 ctgaatctgc tgcaagaaga caacccggac ctgatcggca gcgagatcgc gcgctggctg
     2401 tcgacgctcg agatttccgg caaggatcca ggcaagccca tccccaaccc cctgctgggc
     2461 ctggacagca ccaaagacaa tactgtgcct ctgaagctga tcgctctcct ggctaatggc
     2521 gagttccata gtggcgaaca gctgggagaa accctgggca tgtccagggc cgctatcaac
     2581 aagcacattc agactctgcg cgactggggc gtggacgtgt tcaccgtgcc cggaaagggc
     2641 tactctctgc ccgagcctat cccgctgctg aacgctaaac agattctggg acagctggac
     2701 ggcgggagcg tggcagtcct gcctgtggtc gactccacca atcagtacct gctggatcga
     2761 atcggcgagc tgaagagtgg ggatgcttgc attgcagaat atcagcaggc agggagagga
     2821 agcagaggga ggaaatggtt ctctcctttt ggagctaacc tgtacctgag tatgttttgg
     2881 cgcctgaagc ggggaccagc agcaatcggc ctgggcccgg tcatcggaat tgtcatggca
     2941 gaagcgctgc gaaagctggg agcagacaag gtgcgagtca aatggcccaa tgacctgtat
     3001 ctgcaggata gaaagctggc aggcatcctg gtggagctgg ccggaataac aggcgatgct
     3061 gcacagatcg tcattggcgc cgggattaac gtggctatga ggcgcgtgga ggaaagcgtg
     3121 gtcaatcagg gctggatcac actgcaggaa gcagggatta acctggacag gaatactctg
     3181 gccgctacgc tgatccgaga gctgcgggca gccctggaac tgttcgagca ggaaggcctg
     3241 gctccatatc tgccacggtg ggagaagctg gataacttca tcaatagacc cgtgaagctg
     3301 atcattgggg acaaagagat tttcgggatt agccggggga ttgataaaca gggagccctg
     3361 ctgctggaac aggacggagt tatcaaaccc tggatgggcg gagaaatcag tctgcggtct
     3421 gccgaaaagt aatgatctag agggccctat tctatagtgt cacctaaatg ctagagctcg
     3481 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3541 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3601 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3661 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3721 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3781 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3841 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3901 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3961 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4021 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4081 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4141 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4201 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4261 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4321 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4381 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4441 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4501 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4561 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4621 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4681 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4741 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4801 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4861 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4921 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4981 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5041 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5101 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5161 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5221 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5281 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5341 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5401 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5461 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5521 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5581 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5641 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5701 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5761 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5821 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5881 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5941 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6001 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6061 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6121 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6181 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6241 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6301 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6361 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6421 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6481 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6541 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6601 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6661 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6721 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6781 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6841 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6901 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6961 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7021 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7081 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7141 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7201 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7261 tcagctccgg
//
