LOCUS       sequence-301565-        6019 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hepatocyte-specific expression of ER-localized TurboID, N-term HA 
            tag, C-term V5 tag, in AAV expression plasmid. Amp selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6019)
  AUTHORS   Wei W, Riley NM, Yang AC, Kim JT, Terrell SM, Li VL, 
            Garcia-Contreras M, Bertozzi CR, Long JZ
  TITLE     Cell type-selective secretome profiling in vivo.
  JOURNAL   Nat Chem Biol. 2021 Mar;17(3):326-334. doi: 
            10.1038/s41589-020-00698-y. Epub 2020 Nov 16.
  PUBMED    33199915
REFERENCE   2  (bases 1 to 6019)
  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: 
            "10.1038/s41589-020-00698-y"; journalName: "Nat Chem Biol"; date: 
            "2021-03"; volume: "17"; issue: "3"; pages: "326-334"
COMMENT     Sequence Label: pAAV-TBG-ER-TurboID
FEATURES             Location/Qualifiers
     source          1..6019
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..410
                     /label=TBG promoter
                     /note="promoter from the human thyroxine-binding globulin 
                     gene (Yan et al., 2012)"
     intron          470..602
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     585..604
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      619..628
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             685..711
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             718..1674
                     /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             1681..1722
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1803..1823)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2221..2232)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2299..2316)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2305..2512
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2600..2729
                     /label=AAV2 ITR
     primer_bind     complement(2748..2765)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2748..2764)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2757..2779)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2906..3361
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2993..3012)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3203..3224
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3387..3491
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3492..3560,3561..4352)
                     /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      4523..5111
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5012..5031
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5399..5420
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5435..5440,5441..5458,5459..5465)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5473..5489
                     /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)."
     primer_bind     5478..5500
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5497..5513
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5497..5513
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   5551..5680
                     /label=AAV2 ITR
ORIGIN
        1 tgcatgtata atttctacag aacctattag aaaggatcac ccagcctctg cttttgtaca
       61 actttccctt aaaaaactgc caattccact gctgtttggc ccaatagtga gaactttttc
      121 ctgctgcctc ttggtgcttt tgcctatggc ccctattctg cctgctgaag acactcttgc
      181 cagcatggac ttaaacccct ccagctctga caatcctctt tctcttttgt tttacatgaa
      241 gggtctggca gccaaagcaa tcactcaaag ttcaaacctt atcatttttt gctttgttcc
      301 tcttggcctt ggttttgtac atcagctttg aaaataccat cccagggtta atgctggggt
      361 taatttataa ctaagagtgc tctagttttg caatacagga catgctataa aaatggaaag
      421 atgttgcttt ctgagagact gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      481 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      541 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
      601 aggtgtccag gcggccgcgc caccatggag acagacacac tcctgtgggt gctgctgctc
      661 tgggttccag gttccactgg tgactaccca tacgatgttc cagattacgc tgctagcaaa
      721 gacaatactg tgcctctgaa gctgatcgct ctcctggcta atggcgagtt ccatagtggc
      781 gaacagctgg gagaaaccct gggcatgtcc agggccgcta tcaacaagca cattcagact
      841 ctgcgcgact ggggcgtgga cgtgttcacc gtgcccggaa agggctactc tctgcccgag
      901 cctatcccgc tgctgaacgc taaacagatt ctgggacagc tggacggcgg gagcgtggca
      961 gtcctgcctg tggtcgactc caccaatcag tacctgctgg atcgaatcgg cgagctgaag
     1021 agtggggatg cttgcattgc agaatatcag caggcaggga gaggaagcag agggaggaaa
     1081 tggttctctc cttttggagc taacctgtac ctgagtatgt tttggcgcct gaagcgggga
     1141 ccagcagcaa tcggcctggg cccggtcatc ggaattgtca tggcagaagc gctgcgaaag
     1201 ctgggagcag acaaggtgcg agtcaaatgg cccaatgacc tgtatctgca ggatagaaag
     1261 ctggcaggca tcctggtgga gctggccgga ataacaggcg atgctgcaca gatcgtcatt
     1321 ggcgccggga ttaacgtggc tatgaggcgc gtggaggaaa gcgtggtcaa tcagggctgg
     1381 atcacactgc aggaagcagg gattaacctg gacaggaata ctctggccgc tacgctgatc
     1441 cgagagctgc gggcagccct ggaactgttc gagcaggaag gcctggctcc atatctgcca
     1501 cggtgggaga agctggataa cttcatcaat agacccgtga agctgatcat tggggacaaa
     1561 gagattttcg ggattagccg ggggattgat aaacagggag ccctgctgct ggaacaggac
     1621 ggagttatca aaccctggat gggcggagaa atcagtctgc ggtctgccga aaagctgcag
     1681 ggcaagccca tccccaaccc cctgctgggc ctggacagca ccaaagatga actctaaaag
     1741 cttggatcca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     1801 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     1861 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     1921 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     1981 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2041 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2101 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     2161 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2221 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     2281 ccgcgtcttc gagatctgcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2341 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2401 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2461 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gactcgagtt
     2521 aagggcgaat tcccgataag gatcttccta gagcatggct acgtagataa gtagcatggc
     2581 gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     2641 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     2701 gcggcctcag tgagcgagcg agcgcgcagc cttaattaac ctaattcact ggccgtcgtt
     2761 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     2821 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     2881 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2941 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3001 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3061 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3121 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3181 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3241 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3301 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     3361 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3421 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3481 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3541 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3601 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3661 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3721 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3781 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3841 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3901 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3961 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4021 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4081 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4141 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4201 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4261 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4321 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4381 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4441 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4501 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4561 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4621 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4681 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4741 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4801 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4861 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4921 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4981 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5041 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5101 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5161 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5221 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5281 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5341 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5401 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     5461 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     5521 acgccagatt taattaaggc cttaattagg ctgcgcgctc gctcgctcac tgaggccgcc
     5581 cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg
     5641 cgcagagagg gagtggccaa ctccatcact aggggttcct tgtagttaat gattaacccg
     5701 ccatgctact tatctacgta gccatgctct aggaagatcg gaattcgccc ttaagctagc
     5761 aggttaattt ttaaaaagca gtcaaaagtc caagtggccc ttggcagcat ttactctctc
     5821 tgtttgctct ggttaataat ctcaggagca caaacattcc agatccaggt taatttttaa
     5881 aaagcagtca aaagtccaag tggcccttgg cagcatttac tctctctgtt tgctctggtt
     5941 aataatctca ggagcacaaa cattccagat ccggcgcgcc agggctggaa gctacctttg
     6001 acatcatttc ctctgcgaa
//
