LOCUS       sequence-397527-        4666 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 4666)
  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 4666)
  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) ORF0
FEATURES             Location/Qualifiers
     source          1..4666
                     /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   1929..2069
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1929..2058
                     /label=AAV2 ITR
     rep_origin      2144..2599
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2231..2250)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2441..2462
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2616..2635)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2735..2757
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2795..2813)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2881..2985
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2986..3054,3055..3846)
                     /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      4017..4605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4506..4525
                     /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 gagcctgtac cgtactcgga agcgtccacg tgcggaccga
     1921 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     1981 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     2041 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     2101 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     2161 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2221 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2281 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2341 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2401 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2461 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     2521 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     2581 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     2641 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     2701 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     2761 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     2821 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     2881 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2941 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3001 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3061 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3121 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3181 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     3241 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3301 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3361 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3421 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3481 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     3541 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3601 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3661 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     3721 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     3781 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3841 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3901 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3961 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4021 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4081 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4141 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4201 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4261 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4321 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4381 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4441 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4501 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4561 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4621 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
