LOCUS       sequence-423189-        6524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses 3xHA-tagged TurboID in the mammalian nucleus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6524)
  AUTHORS   Branon TC, Bosch JA, Sanchez AD, Udeshi ND, Svinkina T, Carr SA, 
            Feldman JL, Perrimon N, Ting AY
  TITLE     Efficient proximity labeling in living cells and organisms with 
            TurboID.
  JOURNAL   Nat Biotechnol. 2018 Aug 20. pii: nbt.4201. doi: 10.1038/nbt.4201.
  PUBMED    30125270
REFERENCE   2  (bases 1 to 6524)
  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 
            Biotechnol. 2018 Aug 20. pii: nbt.4201. doi: 10.1038/nbt.4201."
COMMENT     Sequence Label: 3xHA-TurboID-NLS_pCDNA3
FEATURES             Location/Qualifiers
     source          1..6524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(703..807)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(882..901)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1073..1452
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1453..1656
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1607..1627
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1701..1720
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1701..1719
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1748..1774
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1778..1804
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1808..1834
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1841..2797
                     /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"
     CDS             2816..2836
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2840..2860
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2864..2884
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(2915..2933)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2916..2933)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2953..2970)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2959..3183
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3229..3657
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3316..3335)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3526..3547
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3666..3686)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3671..4000
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3851..3986
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3913..3932
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4067..4861
                     /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(4121..4140)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4731..4750
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5035..5156
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5072..5091)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5126..5145
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5205..5221)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5205..5221)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5218..5240)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5229..5245
                     /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(5253..5259,5260..5277,5278..5283))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5298..5319
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5436..5453)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5607..6195)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5687..5706)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6366..6524,1..633,634..702))
                     /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 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
       61 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
      121 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
      181 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
      241 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
      301 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
      361 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
      421 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
      481 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
      541 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
      601 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
      661 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
      721 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
      781 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
      841 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
      901 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
      961 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     1021 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
     1081 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     1141 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     1201 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     1261 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     1321 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     1381 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     1441 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     1501 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     1561 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     1621 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
     1681 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc ttgcggccgc
     1741 caccatgtac ccgtatgatg ttccggatta cgctggctat ccctacgacg tgcccgacta
     1801 tgccgggtac ccctatgacg tcccagacta cgcagctagc aaagacaata ctgtgcctct
     1861 gaagctgatc gctctcctgg ctaatggcga gttccatagt ggcgaacagc tgggagaaac
     1921 cctgggcatg tccagggccg ctatcaacaa gcacattcag actctgcgcg actggggcgt
     1981 ggacgtgttc accgtgcccg gaaagggcta ctctctgccc gagcctatcc cgctgctgaa
     2041 cgctaaacag attctgggac agctggacgg cgggagcgtg gcagtcctgc ctgtggtcga
     2101 ctccaccaat cagtacctgc tggatcgaat cggcgagctg aagagtgggg atgcttgcat
     2161 tgcagaatat cagcaggcag ggagaggaag cagagggagg aaatggttct ctccttttgg
     2221 agctaacctg tacctgagta tgttttggcg cctgaagcgg ggaccagcag caatcggcct
     2281 gggcccggtc atcggaattg tcatggcaga agcgctgcga aagctgggag cagacaaggt
     2341 gcgagtcaaa tggcccaatg acctgtatct gcaggataga aagctggcag gcatcctggt
     2401 ggagctggcc ggaataacag gcgatgctgc acagatcgtc attggcgccg ggattaacgt
     2461 ggctatgagg cgcgtggagg aaagcgtggt caatcagggc tggatcacac tgcaggaagc
     2521 agggattaac ctggacagga atactctggc cgctacgctg atccgagagc tgcgggcagc
     2581 cctggaactg ttcgagcagg aaggcctggc tccatatctg ccacggtggg agaagctgga
     2641 taacttcatc aatagacccg tgaagctgat cattggggac aaagagattt tcgggattag
     2701 ccgggggatt gataaacagg gagccctgct gctggaacag gacggagtta tcaaaccctg
     2761 gatgggcgga gaaatcagtc tgcggtctgc cgaaaaggaa ttcagcaggg ccgaccccaa
     2821 gaagaagagg aaggtggacc ccaagaagaa gaggaaggtg gaccccaaga agaagaggaa
     2881 ggtgtgactc gagcatgcat ctagagggcc ctattctata gtgtcaccta aatgctagag
     2941 ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3001 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3061 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3121 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3181 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3241 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3301 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3361 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3421 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3481 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3541 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3601 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     3661 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3721 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3781 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3841 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3901 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     3961 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     4021 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4081 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4141 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4201 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4261 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4321 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4381 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4441 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4501 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4561 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4621 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4681 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4741 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4801 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4861 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4921 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4981 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5041 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5101 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5161 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5221 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5281 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5341 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5401 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5461 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5521 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5581 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5641 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5701 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5761 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5821 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5881 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5941 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6001 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6061 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6121 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6181 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6241 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6301 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6361 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6421 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6481 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggc
//
