LOCUS       sequence-483885-        5728 bp    DNA     circular SYN 20-AUG-2025
DEFINITION  AAV vector encoding membrane-tethered seTurboID under the control of
            human synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5728)
  TITLE     Ultrabright Chemical Labeling Enables Rapid Neural Connectivity 
            Profiling in Large Tissue Samples
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5728)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 20, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-hSyn-mem-seTurboID
FEATURES             Location/Qualifiers
     source          1..5728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   6..135
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             652..693
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             700..1656
                     /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             1672..1833
                     /codon_start=1
                     /product="B1 domain of Streptococcal protein G   "
                     /label=GB1
                     /note="effective as a solubilizing fusion partner (Cheng 
                     and Patel, 2004)"
                     /translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD
                     ATKTFTVTE"
     misc_feature    1861..2448
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2331..2342)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2480..2956
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2679..2698)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2996..3136
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2996..3125
                     /label=AAV2 ITR
     rep_origin      3211..3666
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3298..3317)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3508..3529
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3683..3702)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3802..3824
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3862..3880)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3948..4052
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4053..4121,4122..4913)
                     /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      5084..5672
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5573..5592
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
       61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      121 tcactagggg ttcctgcggc cgcacgcgta tgaggcgggg tgggggtgcc tacctgacga
      181 ccgaccccga cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat
      241 cagagagggg gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac
      301 cgcggacagt gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag
      361 gcgcgctgac gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg
      421 tccgcgccgc cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg
      481 ggcgcgacca tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca
      541 gcggaggagt cgtgtcgtgc ctgagagcgc agtcgagaaa ccggtgccac catgctgtgc
      601 tgtatgagaa gaaccaaaca ggttgaaaag aatgatgagg accaaaagat cggcaagccc
      661 atccccaacc ccctgctggg cctggacagc accgctagca aagacaatac tgtgcctctg
      721 aagctgatcg ctctcctggc taatggcgag ttccatagtg gcgaacagct gggagaaacc
      781 ctgggcatgt ccagggccgc tatcaacaag cacattcaga ctctgcgcga ctggggcgtg
      841 gacgtgttca ccgtgcccgg aaagggctac tctctgcccg agcctatccc gctgctgaac
      901 gctaaacaga ttctgggaca gctggacggc gggagcgtgg cagtcctgcc tgtggtcgac
      961 tccaccaatc agtacctgct ggatcgaatc ggcgagctga agagtgggga tgcttgcatt
     1021 gcagaatatc agcaggcagg gagaggaagc agagggagga aatggttctc tccttttgga
     1081 gctaacctgt acctgagtat gttttggcgc ctgaagcggg gaccagcagc aatcggcctg
     1141 ggcccggtca tcggaattgt catggcagaa gcgctgcgaa agctgggagc agacaaggtg
     1201 cgagtcaaat ggcccaatga cctgtatctg caggatagaa agctggcagg catcctggtg
     1261 gagctggccg gaataacagg cgatgctgca cagatcgtca ttggcgccgg gattaacgtg
     1321 gctatgaggc gcgtggagga aagcgtggtc aatcagggct ggatcacact gcaggaagca
     1381 gggattaacc tggacaggaa tactctggcc gctacgctga tccgagagct gcgggcagcc
     1441 ctggaactgt tcgagcagga aggcctggct ccatatctgc cacggtggga gaagctggat
     1501 aacttcatca atagacccgt gaagctgatc attggggaca aagagatttt cgggattagc
     1561 cgggggattg ataaacaggg agccctgctg ctggaacagg acggagttat caaaccctgg
     1621 atgggcggag aaatcagtct gcggtctgcc gaaaagggcg gtggcggatc ttacaagctt
     1681 atcctgaacg gtaaaaccct gaaaggtgaa accaccaccg aagctgttga cgctgctacc
     1741 gcggaaaaag ttttcaaaca gtacgctaac gacaacggtg ttgacggtga atggacctac
     1801 gacgacgcta ccaaaacctt cacggtaacc gaataagaat tcgatatcaa gcttatcgat
     1861 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     1921 cctttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     1981 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     2041 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     2101 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     2161 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     2221 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     2281 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     2341 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     2401 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag
     2461 cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     2521 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     2581 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     2641 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     2701 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     2761 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     2821 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     2881 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     2941 tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa
     3001 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     3061 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     3121 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3181 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     3241 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     3301 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3361 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3421 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3481 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3541 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     3601 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     3661 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     3721 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     3781 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     3841 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     3901 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3961 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4021 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4081 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4141 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4201 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4261 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4321 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4381 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4441 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4501 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4561 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4621 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4681 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4741 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4801 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4861 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4921 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4981 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5041 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5101 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5161 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5221 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5281 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5341 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5401 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5461 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5521 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5581 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5641 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5701 tcctggcctt ttgctggcct tttgctca
//
