LOCUS       sequence-354734-        6510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TVA(TC66T) and optimized Rabies G (oG) protein in a FLEX 
            cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6510)
  TITLE     AAV plasmids to express TVA and optimized Rabies G protein
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6510)
  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            "Unpublished"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: AAV-DIO-TC66T-2A-oG
FEATURES             Location/Qualifiers
     source          1..6510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   37..166
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        194..641
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      648..657
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    663..696
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    747..780
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             complement(2362..2418)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     protein_bind    complement(2868..2901)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(2952..2985)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     misc_feature    2987..3575
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3040..3060)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3458..3469)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3583..3807
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3809..3949
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3809..3938
                     /label=AAV2 ITR
     rep_origin      4024..4479
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4111..4130)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4321..4342
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4496..4515)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4615..4637
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4675..4693)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4761..4865
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4866..4934,4935..5726)
                     /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"
     primer_bind     complement(5084..5103)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5897..6485
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6386..6405
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg cgcgctcgct
       61 cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
      121 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcta gactgcagag
      181 ggccctgcgt atgagtgcaa gtgggtttta ggaccaggat gaggcggggt gggggtgcct
      241 acctgacgac cgaccccgac ccactggaca agcacccaac ccccattccc caaattgcgc
      301 atcccctatc agagaggggg aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag
      361 cttcagcacc gcggacagtg ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca
      421 gcactgaagg cgcgctgacg tcactcgccg gtcccccgca aactcccctt cccggccacc
      481 ttggtcgcgt ccgcgccgcc gccggcccag ccggaccgca ccacgcgagg cgcgagatag
      541 gggggcacgg gcgcgaccat ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg
      601 cggtgggcag cggaggagtc gtgtcgtgcc tgagagcgca gtcagccgcc accatggtta
      661 atataacttc gtataatgta tgctatacga agttatgcag aatggtagct ggattgtagc
      721 tgctattagc aatatgaaac ctcttaataa cttcgtataa agtatcctat acgaagttat
      781 ggtaccttag agccgtgtct cgcccccaga tttatgactt tcccaactgc tgatgatttt
      841 accgctctgg ggtgtgacag acacttctct accggtcccc ctcagattgt gctgagttgg
      901 ctcggagcgg ttcactcgtc gccagcaggt catcaggaaa atgatcagca tgagagcagt
      961 cagtgctcca gcggagagca gcacgtactt gccccaatta ggcagtccga gatcgacgcc
     1021 actgatccgt tcgtgcacgt cgggcaggtg gacctccaca aaatcttcgg cctcgtcgcc
     1081 gtttttgaac actgtgcttg ggtctgcgag aggatgcatc aggggaatga cgcttgacac
     1141 gagcagttcc atgtgctgct ggagcagact agactgcatc tcggggatca gcacgttacc
     1201 atctggcccc agaatgatac cattaaagaa gaccccgttc acatgtgggt gacatcgacc
     1261 cccgactcgc aggcatccct tactaggaat gatttcattc caggttctca cagatttata
     1321 atgggcgtct gcctccatca gtgtcttgtt aaagatggtg tatgcttttc cgaagccagg
     1381 gacgagcttc ctcaggtggc tcagcctcct gaagctcacg ctcttggtag tcataattga
     1441 ctccagagca tcgagacatt cctccctttt cttcacgagt tcctcgacca ccagatgttc
     1501 gatctcatcg cttcggaagt cgtgcagatt cacgagctgc ccaggggggc accacttagt
     1561 ctcgttagat gtctgcatag cgacccaggt tccgtccatc agccggagtc ccagcacgcc
     1621 acagagtttc agcttgcagg cacctttgag gctcttatac agccctcttt catccacaaa
     1681 tccacaagtc tcgctgcctt ttgaggccct cttgcccctg ctgttggtga agatgtcgca
     1741 ggacatcccc agccttggat tctcaggcat ccagattgta taatcatggt tggtggaaca
     1801 gtaagtggag ctgacggcca ctccggagca gtttccgccg ggaaacactg gactgtggag
     1861 agatcggtcg taggggtcca ggtcggcgac agatggggaa atgatgacca ggctctcttt
     1921 tgtggtcttc acagtgcgca gccaatgata gtctgggtaa ggattgtgca ggctttcctc
     1981 atagcggggg tcgccagcca tcttccagtt gtaagctgcc cgacaggcgt ctggtgttgg
     2041 ccgaaagtgt tttctcttga aagttgtggt gacatatccc acaaaatttg tgtaggtttc
     2101 tgcctcggtg accacaccag tgcatgtgaa cccgttcatc ttaatagcag agatatagcc
     2161 cactttcagc tccatgtagg agaatccgct caggttggta cagccttcgt cctcgaccac
     2221 gagattgtta gggcatgaca gatggtgaat atcgatagga ctccaggggc ccagcttgtc
     2281 ggggatagtg tagataggga atttaccaaa gcagagtggg aagacgagca gagggacgaa
     2341 gaggagagcc tgtgggacca tgggtccagg gttctcctcc acgtctccag cctgcttcag
     2401 caggctgaag ttagtagctc cggatccccc accccccttg gatgcgcttt caagactgaa
     2461 gatgtcagac ctgctttttg ctttggactt cccccatgca gcgataccac ccacagctac
     2521 caggcagcac aggagcactg cagtgatcag catccacatg cggccgtgat tcctggctgg
     2581 cagagcacgt ccaggagcag gggcagtggg agcctctgtg ccgttgtccg tgggcaccgc
     2641 ggggatcgcg ctggtcccgc agccccaggt gtcccgcccg tcgtcgcagt cggggtgtcc
     2701 gtcgcacagc cagtcctgcg ggtaacactc cccgtgggca ccgggcggct ccgagcagcg
     2761 gaactgaccg ggggggcaac gggacaaaga accgttaccg gacccgttac cggtcacgtt
     2821 accgggcagc agcagcagca gcagcgcggg cagcagccgc gcacctaata acttcgtata
     2881 gcatacatta tacgaagtta tttgccttaa cccagaaatt atcactgtta ttctttagaa
     2941 tggtgcaaag aataacttcg tataggatac tttatacgaa gttatgaatc aacctctgga
     3001 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3061 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3121 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3181 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3241 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3301 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3361 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     3421 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3481 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3541 gacgagtcgg atctcccttt gggccgcctc cccgcctgga tcctgtgcct tctagttgcc
     3601 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     3661 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3721 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     3781 atgctgggga tgcggtgggc tctatggcag gaacccctag tgatggagtt ggccactccc
     3841 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     3901 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     3961 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     4021 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4081 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4141 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4201 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     4261 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4321 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     4381 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     4441 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     4501 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     4561 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4621 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     4681 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     4741 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4801 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4861 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4921 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4981 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5041 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5101 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5161 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5221 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5281 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5341 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5401 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5461 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5521 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5581 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5641 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5701 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5761 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5821 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5881 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5941 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6001 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6061 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6121 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6181 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6241 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6301 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6361 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6421 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6481 ggaaaaacgc cagcaacgcg gcctttttac
//
