LOCUS       sequence-354735-        6543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TVA 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 6543)
  TITLE     AAV plasmids to express TVA and optimized Rabies G protein
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6543)
  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            "Unpublished"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-TVA-2A-oG
FEATURES             Location/Qualifiers
     source          1..6543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      358..367
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    373..406
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    457..490
                     /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(2072..2128)
                     /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(2611..2644)
                     /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(2695..2728)
                     /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    2730..3318
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2783..2803)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3201..3212)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3326..3550
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3552..3692
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3552..3681
                     /label=AAV2 ITR
     rep_origin      3767..4222
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3854..3873)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4064..4085
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4239..4258)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4358..4380
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4418..4436)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4504..4608
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4609..4677,4678..5469)
                     /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(4827..4846)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5640..6228
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6129..6148
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6290..6419
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        join(6447..6543,1..351)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 caaattgcgc atcccctatc agagaggggg aggggaaaca ggatgcggcg aggcgcgtgc
       61 gcactgccag cttcagcacc gcggacagtg ccttcgcccc cgcctggcgg cgcgcgccac
      121 cgccgcctca gcactgaagg cgcgctgacg tcactcgccg gtcccccgca aactcccctt
      181 cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag ccggaccgca ccacgcgagg
      241 cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg gcgccggcga ctcagcgctg
      301 cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc tgagagcgca gtcagccgcc
      361 accatggtta atataacttc gtataatgta tgctatacga agttatgcag aatggtagct
      421 ggattgtagc tgctattagc aatatgaaac ctcttaataa cttcgtataa agtatcctat
      481 acgaagttat ggtaccttag agccgtgtct cgcccccaga tttatgactt tcccaactgc
      541 tgatgatttt accgctctgg ggtgtgacag acacttctct accggtcccc ctcagattgt
      601 gctgagttgg ctcggagcgg ttcactcgtc gccagcaggt catcaggaaa atgatcagca
      661 tgagagcagt cagtgctcca gcggagagca gcacgtactt gccccaatta ggcagtccga
      721 gatcgacgcc actgatccgt tcgtgcacgt cgggcaggtg gacctccaca aaatcttcgg
      781 cctcgtcgcc gtttttgaac actgtgcttg ggtctgcgag aggatgcatc aggggaatga
      841 cgcttgacac gagcagttcc atgtgctgct ggagcagact agactgcatc tcggggatca
      901 gcacgttacc atctggcccc agaatgatac cattaaagaa gaccccgttc acatgtgggt
      961 gacatcgacc cccgactcgc aggcatccct tactaggaat gatttcattc caggttctca
     1021 cagatttata atgggcgtct gcctccatca gtgtcttgtt aaagatggtg tatgcttttc
     1081 cgaagccagg gacgagcttc ctcaggtggc tcagcctcct gaagctcacg ctcttggtag
     1141 tcataattga ctccagagca tcgagacatt cctccctttt cttcacgagt tcctcgacca
     1201 ccagatgttc gatctcatcg cttcggaagt cgtgcagatt cacgagctgc ccaggggggc
     1261 accacttagt ctcgttagat gtctgcatag cgacccaggt tccgtccatc agccggagtc
     1321 ccagcacgcc acagagtttc agcttgcagg cacctttgag gctcttatac agccctcttt
     1381 catccacaaa tccacaagtc tcgctgcctt ttgaggccct cttgcccctg ctgttggtga
     1441 agatgtcgca ggacatcccc agccttggat tctcaggcat ccagattgta taatcatggt
     1501 tggtggaaca gtaagtggag ctgacggcca ctccggagca gtttccgccg ggaaacactg
     1561 gactgtggag agatcggtcg taggggtcca ggtcggcgac agatggggaa atgatgacca
     1621 ggctctcttt tgtggtcttc acagtgcgca gccaatgata gtctgggtaa ggattgtgca
     1681 ggctttcctc atagcggggg tcgccagcca tcttccagtt gtaagctgcc cgacaggcgt
     1741 ctggtgttgg ccgaaagtgt tttctcttga aagttgtggt gacatatccc acaaaatttg
     1801 tgtaggtttc tgcctcggtg accacaccag tgcatgtgaa cccgttcatc ttaatagcag
     1861 agatatagcc cactttcagc tccatgtagg agaatccgct caggttggta cagccttcgt
     1921 cctcgaccac gagattgtta gggcatgaca gatggtgaat atcgatagga ctccaggggc
     1981 ccagcttgtc ggggatagtg tagataggga atttaccaaa gcagagtggg aagacgagca
     2041 gagggacgaa gaggagagcc tgtgggacca tgggtccagg gttctcctcc acgtctccag
     2101 cctgcttcag caggctgaag ttagtagctc cggatccgga gaacaagtct gcctggctct
     2161 tgtcaggcac cagcagctcc ttggatgcgc tttcaagact gaagatgtca gacctgcttt
     2221 ttgctttgga cttcccccat gcagcgatac cacccacagc taccaggcag cacaggagca
     2281 ctgcagtgat cagcatccac atgcggccgt gattcctggc tggcagagca cgtccaggag
     2341 caggggcagt gggagcctct gtgccgttgt ccgtgggcac cgcggggatc gcgctggtcc
     2401 cgcagcccca ctcgtcccgc ccgtcgtcgc agtcggggtg tccgtcgcac agccagtcct
     2461 gcgggtaaca ctccccgtgg gcaccgggcg gctccgagca gcggaactga ccgggggggc
     2521 aacgggacaa agaaccgtta ccggacccgt taccggtcac gttaccgggc agcagcagca
     2581 gcagcagcgc gggcagcagc cgcgcaccta ataacttcgt atagcataca ttatacgaag
     2641 ttatttgcct taacccagaa attatcactg ttattcttta gaatggtgca aagaataact
     2701 tcgtatagga tactttatac gaagttatga atcaacctct ggattacaaa atttgtgaaa
     2761 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     2821 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     2881 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     2941 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3001 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3061 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3121 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     3181 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     3241 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     3301 tttgggccgc ctccccgcct ggatcctgtg ccttctagtt gccagccatc tgttgtttgc
     3361 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3421 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3481 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3541 ggctctatgg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     3601 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     3661 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     3721 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     3781 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3841 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3901 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3961 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     4021 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4081 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
     4141 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     4201 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     4261 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     4321 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4381 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     4441 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     4501 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4561 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4621 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4681 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4741 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4801 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4861 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4921 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4981 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5041 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5101 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5161 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5221 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5281 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5341 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5401 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5461 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5521 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5581 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5641 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5701 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5761 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5821 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5881 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5941 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6001 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6061 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6121 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6181 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6241 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtc ctgcaggcag
     6301 ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc
     6361 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg
     6421 ctagactgca gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg
     6481 ggtgggggtg cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt
     6541 ccc
//
