LOCUS       sequence-301563-        5947 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hepatocyte-specific expression of cytosolic TurboID, N-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 5947)
  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 5947)
  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-Cyto-TurboID
FEATURES             Location/Qualifiers
     source          1..5947
                     /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"
     CDS             622..663
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             670..1626
                     /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             1633..1659
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(1731..1751)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2149..2160)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2227..2244)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2233..2440
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2528..2657
                     /label=AAV2 ITR
     primer_bind     complement(2676..2693)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2676..2692)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2685..2707)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2834..3289
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2921..2940)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3131..3152
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3315..3419
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3420..3488,3489..4280)
                     /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      4451..5039
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4940..4959
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5327..5348
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5363..5368,5369..5386,5387..5393)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5401..5417
                     /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     5406..5428
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5425..5441
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5425..5441
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   5479..5608
                     /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 gcggccgcat gggcaagccc atccccaacc ccctgctggg cctggacagc
      661 accgctagca aagacaatac tgtgcctctg aagctgatcg ctctcctggc taatggcgag
      721 ttccatagtg gcgaacagct gggagaaacc ctgggcatgt ccagggccgc tatcaacaag
      781 cacattcaga ctctgcgcga ctggggcgtg gacgtgttca ccgtgcccgg aaagggctac
      841 tctctgcccg agcctatccc gctgctgaac gctaaacaga ttctgggaca gctggacggc
      901 gggagcgtgg cagtcctgcc tgtggtcgac tccaccaatc agtacctgct ggatcgaatc
      961 ggcgagctga agagtgggga tgcttgcatt gcagaatatc agcaggcagg gagaggaagc
     1021 agagggagga aatggttctc tccttttgga gctaacctgt acctgagtat gttttggcgc
     1081 ctgaagcggg gaccagcagc aatcggcctg ggcccggtca tcggaattgt catggcagaa
     1141 gcgctgcgaa agctgggagc agacaaggtg cgagtcaaat ggcccaatga cctgtatctg
     1201 caggatagaa agctggcagg catcctggtg gagctggccg gaataacagg cgatgctgca
     1261 cagatcgtca ttggcgccgg gattaacgtg gctatgaggc gcgtggagga aagcgtggtc
     1321 aatcagggct ggatcacact gcaggaagca gggattaacc tggacaggaa tactctggcc
     1381 gctacgctga tccgagagct gcgggcagcc ctggaactgt tcgagcagga aggcctggct
     1441 ccatatctgc cacggtggga gaagctggat aacttcatca atagacccgt gaagctgatc
     1501 attggggaca aagagatttt cgggattagc cgggggattg ataaacaggg agccctgctg
     1561 ctggaacagg acggagttat caaaccctgg atgggcggag aaatcagtct gcggtctgcc
     1621 gaaaagctgc agctgcctcc cctggagcgc ctgaccctgg actaaaagct tggatccaat
     1681 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     1741 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     1801 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     1861 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     1921 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     1981 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     2041 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     2101 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     2161 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcga
     2221 gatctgcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2281 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2341 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2401 agggggagga ttgggaagac aatagcaggc atgctgggga ctcgagttaa gggcgaattc
     2461 ccgataagga tcttcctaga gcatggctac gtagataagt agcatggcgg gttaatcatt
     2521 aactacaagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     2581 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     2641 agcgagcgag cgcgcagcct taattaacct aattcactgg ccgtcgtttt acaacgtcgt
     2701 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     2761 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     2821 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2881 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2941 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3001 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3061 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3121 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3181 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3241 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     3301 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3361 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3421 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3481 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3541 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3601 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3661 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3721 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3781 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3841 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3901 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3961 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4021 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4081 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4141 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4201 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4261 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4321 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4381 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4441 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4501 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4561 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4621 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4681 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4741 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4801 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4861 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4921 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4981 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5041 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5101 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     5161 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     5221 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     5281 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     5341 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     5401 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccagattta
     5461 attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
     5521 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
     5581 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
     5641 tctacgtagc catgctctag gaagatcgga attcgccctt aagctagcag gttaattttt
     5701 aaaaagcagt caaaagtcca agtggccctt ggcagcattt actctctctg tttgctctgg
     5761 ttaataatct caggagcaca aacattccag atccaggtta atttttaaaa agcagtcaaa
     5821 agtccaagtg gcccttggca gcatttactc tctctgtttg ctctggttaa taatctcagg
     5881 agcacaaaca ttccagatcc ggcgcgccag ggctggaagc tacctttgac atcatttcct
     5941 ctgcgaa
//
