LOCUS       sequence-300792-        6160 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hepatocyte-specific expression of secretory pathway 
            membrane-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 6160)
  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 6160)
  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-mem-TurboID
FEATURES             Location/Qualifiers
     source          1..6160
                     /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"
     sig_peptide     625..684
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             688..714
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             721..1677
                     /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             1684..1725
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1729..1875
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(1944..1964)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2362..2373)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2440..2457)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2446..2653
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2741..2870
                     /label=AAV2 ITR
     primer_bind     complement(2889..2906)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2889..2905)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2898..2920)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3047..3502
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3134..3153)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3344..3365
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3528..3632
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3633..3701,3702..4493)
                     /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      4664..5252
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5153..5172
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5540..5561
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5576..5581,5582..5599,5600..5606)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5614..5630
                     /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     5619..5641
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5638..5654
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5638..5654
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   5692..5821
                     /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 tcctgctatg ggtgctgctg
      661 ctctgggttc caggttccac tggtgactac ccatacgatg ttccagatta cgctgctagc
      721 aaagacaata ctgtgcctct gaagctgatc gctctcctgg ctaatggcga gttccatagt
      781 ggcgaacagc tgggagaaac cctgggcatg tccagggccg ctatcaacaa gcacattcag
      841 actctgcgcg actggggcgt ggacgtgttc accgtgcccg gaaagggcta ctctctgccc
      901 gagcctatcc cgctgctgaa cgctaaacag attctgggac agctggacgg cgggagcgtg
      961 gcagtcctgc ctgtggtcga ctccaccaat cagtacctgc tggatcgaat cggcgagctg
     1021 aagagtgggg atgcttgcat tgcagaatat cagcaggcag ggagaggaag cagagggagg
     1081 aaatggttct ctccttttgg agctaacctg tacctgagta tgttttggcg cctgaagcgg
     1141 ggaccagcag caatcggcct gggcccggtc atcggaattg tcatggcaga agcgctgcga
     1201 aagctgggag cagacaaggt gcgagtcaaa tggcccaatg acctgtatct gcaggataga
     1261 aagctggcag gcatcctggt ggagctggcc ggaataacag gcgatgctgc acagatcgtc
     1321 attggcgccg ggattaacgt ggctatgagg cgcgtggagg aaagcgtggt caatcagggc
     1381 tggatcacac tgcaggaagc agggattaac ctggacagga atactctggc cgctacgctg
     1441 atccgagagc tgcgggcagc cctggaactg ttcgagcagg aaggcctggc tccatatctg
     1501 ccacggtggg agaagctgga taacttcatc aatagacccg tgaagctgat cattggggac
     1561 aaagagattt tcgggattag ccgggggatt gataaacagg gagccctgct gctggaacag
     1621 gacggagtta tcaaaccctg gatgggcgga gaaatcagtc tgcggtctgc cgaaaagctg
     1681 cagggcaagc ccatccccaa ccccctgctg ggcctggaca gcaccaatgc tgtgggccag
     1741 gacacgcagg aggtcatcgt ggtgccacac tccttgccct ttaaggtggt ggtgatctca
     1801 gccatcctgg ccctggtggt gctcaccatc atctccctta tcatcctcat catgctttgg
     1861 cagaagaagc cacgttaaaa gcttggatcc aatcaacctc tggattacaa aatttgtgaa
     1921 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     1981 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     2041 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     2101 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     2161 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     2221 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     2281 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     2341 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     2401 ctgccggctc tgcggcctct tccgcgtctt cgagatctgc ctcgactgtg ccttctagtt
     2461 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2521 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2581 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2641 ggcatgctgg ggactcgagt taagggcgaa ttcccgataa ggatcttcct agagcatggc
     2701 tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag
     2761 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     2821 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ccttaattaa
     2881 cctaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     2941 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     3001 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     3061 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3121 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3181 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3241 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3301 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3361 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3421 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     3481 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3541 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3601 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3661 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3721 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3781 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3841 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3901 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3961 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4021 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4081 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4141 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4201 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4261 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4321 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4381 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4441 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4501 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4561 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4621 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4681 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4741 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4801 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4861 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4921 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4981 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5041 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5101 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5161 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5221 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5281 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5341 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     5401 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     5461 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     5521 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     5581 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     5641 gaaacagcta tgaccatgat tacgccagat ttaattaagg ccttaattag gctgcgcgct
     5701 cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg
     5761 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
     5821 ttgtagttaa tgattaaccc gccatgctac ttatctacgt agccatgctc taggaagatc
     5881 ggaattcgcc cttaagctag caggttaatt tttaaaaagc agtcaaaagt ccaagtggcc
     5941 cttggcagca tttactctct ctgtttgctc tggttaataa tctcaggagc acaaacattc
     6001 cagatccagg ttaattttta aaaagcagtc aaaagtccaa gtggcccttg gcagcattta
     6061 ctctctctgt ttgctctggt taataatctc aggagcacaa acattccaga tccggcgcgc
     6121 cagggctgga agctaccttt gacatcattt cctctgcgaa
//
