LOCUS       sequence-299816-        6076 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle-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 6076)
  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 6076)
  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-tMCK-ER-TurboID
FEATURES             Location/Qualifiers
     source          1..6076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          1004..1136
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1119..1138
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1153..1162
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1159..1218
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             1222..1248
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1255..2211
                     /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             2218..2259
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2340..2360)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2758..2769)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2836..2853)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2842..3049
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3137..3266
                     /label=AAV2 ITR
     primer_bind     complement(3285..3302)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3285..3301)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3294..3316)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3443..3898
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3530..3549)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3740..3761
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3924..4028
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4029..4097,4098..4889)
                     /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      5060..5648
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5549..5568
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5936..5957
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5972..5977,5978..5995,5996..6002)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6010..6026
                     /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     6015..6037
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6034..6050
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6034..6050
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ccttaattag gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcggc
       61 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctt
      121 gtagttaatg attaacccgc catgctactt atctacgtag ccatgctcta ggaagatcgg
      181 aattcgccct taagctagcc actacgggtc taggctgccc atgtaaggag gcaaggcctg
      241 gggacacccg agatgcctgg ttataattaa ccccaacacc tgctgccccc ccccccccaa
      301 cacctgctgc ctgagcctga gcggttaccc caccccggtg cctgggtctt aggctctgta
      361 caccatggag gagaagctcg ctctaaaaat aaccctgtcc ctggtgggcc cactacgggt
      421 ctaggctgcc catgtaagga ggcaaggcct ggggacaccc gagatgcctg gttataatta
      481 accccaacac ctgctgcccc ccccccccca acacctgctg cctgagcctg agcggttacc
      541 ccaccccggt gcctgggtct taggctctgt acaccatgga ggagaagctc gctctaaaaa
      601 taaccctgtc cctggtgggc cactacgggt ctaggctgcc catgtaagga ggcaaggcct
      661 ggggacaccc gagatgcctg gttataatta accccaacac ctgctgcccc ccccccccca
      721 acacctgctg cctgagcctg agcggttacc ccaccccggt gcctgggtct taggctctgt
      781 acaccatgga ggagaagctc gctctaaaaa taaccctgtc cctggtgggc ccctccctgg
      841 ggacagcccc tcctggctag tcacaccctg taggctcctc tatataaccc aggggcacag
      901 gggctgcccc cgggtcacgg ggatcctcta gacctccgaa cggtaagagc ctagcatgta
      961 gaactggtta ccctgcagaa gttggtcgtg aggcactggg caggtaagta tcaaggttac
     1021 aagacaggtt taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg
     1081 tttctgatag gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt
     1141 ccaggcggcc gcgccaccat ggagacagac acactcctgc tatgggtgct gctgctctgg
     1201 gttccaggtt ccactggtga ctacccatac gatgttccag attacgctgc tagcaaagac
     1261 aatactgtgc ctctgaagct gatcgctctc ctggctaatg gcgagttcca tagtggcgaa
     1321 cagctgggag aaaccctggg catgtccagg gccgctatca acaagcacat tcagactctg
     1381 cgcgactggg gcgtggacgt gttcaccgtg cccggaaagg gctactctct gcccgagcct
     1441 atcccgctgc tgaacgctaa acagattctg ggacagctgg acggcgggag cgtggcagtc
     1501 ctgcctgtgg tcgactccac caatcagtac ctgctggatc gaatcggcga gctgaagagt
     1561 ggggatgctt gcattgcaga atatcagcag gcagggagag gaagcagagg gaggaaatgg
     1621 ttctctcctt ttggagctaa cctgtacctg agtatgtttt ggcgcctgaa gcggggacca
     1681 gcagcaatcg gcctgggccc ggtcatcgga attgtcatgg cagaagcgct gcgaaagctg
     1741 ggagcagaca aggtgcgagt caaatggccc aatgacctgt atctgcagga tagaaagctg
     1801 gcaggcatcc tggtggagct ggccggaata acaggcgatg ctgcacagat cgtcattggc
     1861 gccgggatta acgtggctat gaggcgcgtg gaggaaagcg tggtcaatca gggctggatc
     1921 acactgcagg aagcagggat taacctggac aggaatactc tggccgctac gctgatccga
     1981 gagctgcggg cagccctgga actgttcgag caggaaggcc tggctccata tctgccacgg
     2041 tgggagaagc tggataactt catcaataga cccgtgaagc tgatcattgg ggacaaagag
     2101 attttcggga ttagccgggg gattgataaa cagggagccc tgctgctgga acaggacgga
     2161 gttatcaaac cctggatggg cggagaaatc agtctgcggt ctgccgaaaa gctgcagggc
     2221 aagcccatcc ccaaccccct gctgggcctg gacagcacca aagatgaact ctaaaagctt
     2281 ggatccaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     2341 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     2401 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     2461 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     2521 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     2581 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     2641 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     2701 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     2761 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     2821 cgtcttcgag atctgcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     2881 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     2941 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3001 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggac tcgagttaag
     3061 ggcgaattcc cgataaggat cttcctagag catggctacg tagataagta gcatggcggg
     3121 ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3181 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3241 gcctcagtga gcgagcgagc gcgcagcctt aattaaccta attcactggc cgtcgtttta
     3301 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     3361 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     3421 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3481 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3541 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3601 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3661 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3721 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3781 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3841 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     3901 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3961 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4021 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4081 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4141 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4201 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4261 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4321 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4381 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4441 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4501 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4561 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4621 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4681 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4741 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4801 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4861 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4921 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4981 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5041 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5101 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5161 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5221 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5281 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5341 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5401 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5461 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5521 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5581 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5641 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5701 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     5761 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     5821 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     5881 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     5941 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     6001 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     6061 ccagatttaa ttaagg
//
