LOCUS       sequence-397528-        4667 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 4667)
  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 4667)
  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) ORF2
FEATURES             Location/Qualifiers
     source          1..4667
                     /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   1930..2070
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1930..2059
                     /label=AAV2 ITR
     rep_origin      2145..2600
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2232..2251)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2442..2463
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2617..2636)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2736..2758
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2796..2814)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2882..2986
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2987..3055,3056..3847)
                     /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      4018..4606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4507..4526
                     /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 tgtctttcct cgagcctgta ccgtactcgg aagcgtccac gtgcggaccg
     1921 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     1981 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     2041 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     2101 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     2161 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2221 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2281 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2341 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2401 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2461 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     2521 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     2581 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     2641 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     2701 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     2761 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     2821 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     2881 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2941 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3001 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3061 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3121 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3181 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     3241 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3301 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3361 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3421 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3481 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     3541 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3601 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3661 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3721 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3781 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3841 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3901 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3961 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4021 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4081 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4141 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4201 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4261 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4321 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4381 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4441 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4501 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4561 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4621 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
