LOCUS       sequence-397392-        4668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tagging endogenously expressed proteins with TurboID-HA, C-terminal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4668)
  AUTHORS   Gao Y, Shonai D, Trn M, Zhao J, Soderblom EJ, Garcia-Moreno SA, 
            Gersbach CA, Wetsel WC, Dawson G, Velmeshev D, Jiang YH, Sloofman 
            LG, Buxbaum JD, Soderling SH
  TITLE     Proximity analysis of native proteomes reveals phenotypic modifiers 
            in a mouse model of autism and related neurodevelopmental 
            conditions.
  JOURNAL   Nat Commun. 2024 Aug 9;15(1):6801. doi: 10.1038/s41467-024-51037-x.
  PUBMED    39122707
REFERENCE   2  (bases 1 to 4668)
  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 
            Commun."; date: "2024-08-9"; volume: "15(1):6801. doi"; pages: " 
            10.1038/s41467-024-51037-x"
COMMENT     Sequence Label: pHiUGE TurboID-HA donor (all-in-one) ORF1
FEATURES             Location/Qualifiers
     source          1..4668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        145..393
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        414..489
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        501..749
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        768..843
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(888..914)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(921..1877)
                     /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"
     repeat_region   1931..2071
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1931..2060
                     /label=AAV2 ITR
     rep_origin      2146..2601
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2233..2252)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2443..2464
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2618..2637)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2737..2759
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2797..2815)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2883..2987
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2988..3056,3057..3848)
                     /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"
     rep_origin      4019..4607
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4508..4527
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgagggc ctatttccca tgattccttc atatttgcat
      181 atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata
      241 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
      301 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
      361 gctttatata tcttgtggaa aggacgaaac accgacgctt ccgagtacgg tacgttttag
      421 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
      481 agtcggtgct ttttttctag gagggcctat ttcccatgat tccttcatat ttgcatatac
      541 gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag
      601 tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat
      661 gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt
      721 tatatatctt gtggaaagga cgaaacaccg gaagagcgag ctcttctgtt ttagagctag
      781 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
      841 tgcttttttt ctagacctgt accgtactcg gaagcgtcgg atccttatgc gtaatccggt
      901 acatcgtaag ggtagaattc cttttcggca gaccgcagac tgatttctcc gcccatccag
      961 ggtttgataa ctccgtcctg ttccagcagc agggctccct gtttatcaat cccccggcta
     1021 atcccgaaaa tctctttgtc cccaatgatc agcttcacgg gtctattgat gaagttatcc
     1081 agcttctccc accgtggcag atatggagcc aggccttcct gctcgaacag ttccagggct
     1141 gcccgcagct ctcggatcag cgtagcggcc agagtattcc tgtccaggtt aatccctgct
     1201 tcctgcagtg tgatccagcc ctgattgacc acgctttcct ccacgcgcct catagccacg
     1261 ttaatcccgg cgccaatgac gatctgtgca gcatcgcctg ttattccggc cagctccacc
     1321 aggatgcctg ccagctttct atcctgcaga tacaggtcat tgggccattt gactcgcacc
     1381 ttgtctgctc ccagctttcg cagcgcttct gccatgacaa ttccgatgac cgggcccagg
     1441 ccgattgctg ctggtccccg cttcaggcgc caaaacatac tcaggtacag gttagctcca
     1501 aaaggagaga accatttcct ccctctgctt cctctccctg cctgctgata ttctgcaatg
     1561 caagcatccc cactcttcag ctcgccgatt cgatccagca ggtactgatt ggtggagtcg
     1621 accacaggca ggactgccac gctcccgccg tccagctgtc ccagaatctg tttagcgttc
     1681 agcagcggga taggctcggg cagagagtag ccctttccgg gcacggtgaa cacgtccacg
     1741 ccccagtcgc gcagagtctg aatgtgcttg ttgatagcgg ccctggacat gcccagggtt
     1801 tctcccagct gttcgccact atggaactcg ccattagcca ggagagcgat cagcttcaga
     1861 ggcacagtat tgtctttctc tcgagcctgt accgtactcg gaagcgtcca cgtgcggacc
     1921 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     1981 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     2041 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     2101 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     2161 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2221 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2281 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2341 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     2401 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2461 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     2521 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     2581 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     2641 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     2701 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     2761 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     2821 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     2881 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2941 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3001 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3061 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3121 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3181 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3241 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3301 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3361 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3421 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3481 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3541 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3601 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3661 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3721 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3781 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3841 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     3901 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     3961 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4021 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4081 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4141 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4201 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4261 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4321 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4381 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4441 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4501 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4561 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4621 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
