LOCUS       sequence-299815-        6001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle-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 6001)
  AUTHORS   Long J
  TITLE     Jonathan Long Lab
  JOURNAL   (Unpublished)
REFERENCE   2  (bases 1 to 6001)
  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: 
            "(Unpublished)"
COMMENT     Sequence Label: pAAV-tMCK-Cyto-TurboID
FEATURES             Location/Qualifiers
     source          1..6001
                     /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"
     CDS             1156..1197
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1204..2160
                     /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             2167..2193
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(2265..2285)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2683..2694)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2761..2778)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2767..2974
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3062..3191
                     /label=AAV2 ITR
     primer_bind     complement(3210..3227)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3210..3226)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3219..3241)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3368..3823
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3455..3474)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3665..3686
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3849..3953
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3954..4022,4023..4814)
                     /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      4985..5573
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5474..5493
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5861..5882
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5897..5902,5903..5920,5921..5927)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5935..5951
                     /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     5940..5962
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5959..5975
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5959..5975
                     /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 gcatgggcaa gcccatcccc aaccccctgc tgggcctgga cagcaccgct
     1201 agcaaagaca atactgtgcc tctgaagctg atcgctctcc tggctaatgg cgagttccat
     1261 agtggcgaac agctgggaga aaccctgggc atgtccaggg ccgctatcaa caagcacatt
     1321 cagactctgc gcgactgggg cgtggacgtg ttcaccgtgc ccggaaaggg ctactctctg
     1381 cccgagccta tcccgctgct gaacgctaaa cagattctgg gacagctgga cggcgggagc
     1441 gtggcagtcc tgcctgtggt cgactccacc aatcagtacc tgctggatcg aatcggcgag
     1501 ctgaagagtg gggatgcttg cattgcagaa tatcagcagg cagggagagg aagcagaggg
     1561 aggaaatggt tctctccttt tggagctaac ctgtacctga gtatgttttg gcgcctgaag
     1621 cggggaccag cagcaatcgg cctgggcccg gtcatcggaa ttgtcatggc agaagcgctg
     1681 cgaaagctgg gagcagacaa ggtgcgagtc aaatggccca atgacctgta tctgcaggat
     1741 agaaagctgg caggcatcct ggtggagctg gccggaataa caggcgatgc tgcacagatc
     1801 gtcattggcg ccgggattaa cgtggctatg aggcgcgtgg aggaaagcgt ggtcaatcag
     1861 ggctggatca cactgcagga agcagggatt aacctggaca ggaatactct ggccgctacg
     1921 ctgatccgag agctgcgggc agccctggaa ctgttcgagc aggaaggcct ggctccatat
     1981 ctgccacggt gggagaagct ggataacttc atcaatagac ccgtgaagct gatcattggg
     2041 gacaaagaga ttttcgggat tagccggggg attgataaac agggagccct gctgctggaa
     2101 caggacggag ttatcaaacc ctggatgggc ggagaaatca gtctgcggtc tgccgaaaag
     2161 ctgcagctgc ctcccctgga gcgcctgacc ctggactaaa agcttggatc caatcaacct
     2221 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2281 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2341 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     2401 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     2461 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     2521 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2581 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     2641 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     2701 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgagatctg
     2761 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     2821 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2881 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2941 aggattggga agacaatagc aggcatgctg gggactcgag ttaagggcga attcccgata
     3001 aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac
     3061 aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     3121 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     3181 cgagcgcgca gccttaatta acctaattca ctggccgtcg ttttacaacg tcgtgactgg
     3241 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     3301 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     3361 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3421 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3481 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3541 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3601 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3661 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3721 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3781 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     3841 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3901 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3961 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4021 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4081 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4141 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4201 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4261 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4321 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4381 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4441 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4501 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4561 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4621 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4681 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4741 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4801 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4861 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4921 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4981 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5041 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5101 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5161 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5221 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5281 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5341 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5401 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5461 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5521 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5581 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5641 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     5701 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     5761 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     5821 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     5881 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     5941 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaga tttaattaag
     6001 g
//
