LOCUS       sequence-485436-        7736 bp    DNA     circular SYN 05-SEP-2025
DEFINITION  Encodes the G protein to enable transsynaptic RAV infection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7736)
  AUTHORS   Reardon TR, Murray AJ, Turi GF, Wirblich C, Croce KR, Schnell MJ, 
            Jessell TM, Losonczy A
  TITLE     Rabies Virus CVS-N2c Strain Enhances Retrograde Synaptic Transfer 
            and Neuronal Viability.
  JOURNAL   Neuron. 2016 Jan 19. pii: S0896-6273(16)00005-2. doi: 
            10.1016/j.neuron.2016.01.004.
  PUBMED    26804990
REFERENCE   2  (bases 1 to 7736)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Neuron. 
            2016 Jan 19. pii: S0896-6273(16)00005-2. doi: 
            10.1016/j.neuron.2016.01.004."
COMMENT     Sequence Label: DIO-EF1a-FLEX-H2B-3XHA-P2A-N2c(G)
FEATURES             Location/Qualifiers
     source          1..7736
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1440..1473)
                     /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)."
     protein_bind    complement(1524..1557)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(3154..3210)
                     /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"
     CDS             complement(3220..3309)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             complement(3328..3705)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSSK"
     protein_bind    3721..3754
                     /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)."
     protein_bind    3805..3838
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3863..4451
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3916..3936)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4334..4345)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4483..4959
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4682..4701)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4999..5139
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4999..5128
                     /label=AAV2 ITR
     rep_origin      5214..5669
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5301..5320)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5511..5532
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5686..5705)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5805..5827
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5865..5883)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5951..6055
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6056..6124,6125..6916)
                     /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(6274..6293)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7087..7675
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7576..7595
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa
     1441 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
     1501 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
     1561 gcgccttact tgtacatctt acagtctgat ctcacctcca ctcttatatg attcccatga
     1621 aggtatgact tttccgcttt gggaagtgac tgacacattc ccccctgtcc ctccaaaact
     1681 gcgttgtttc gattctggtc gattggctct tctgcaccat gtcattaggg aaaatatcaa
     1741 caccaggcca atcatggccc ctgcagtcat caatacatac tttccccagt tcgggagacc
     1801 caggtcaacc cctgagatct gtttgtacac atcggggagg tgaacttcaa caaaatcctc
     1861 agcctcatca ccttctttga aaactgtaga agggtctgcc agggggtgca tcagggggat
     1921 aactgaagat tccaacaact ccatatgttg ctggaggagg gatgattgca tctctgggat
     1981 taggacatgg tcgtcaggcc ctaatattat accattgaaa aacaccccgt tcacatgagg
     2041 atggcacctt cctccaactt tcaaacaccc ttttgagggg atgatctcat tccaggtccg
     2101 gactgacttg tagtgagcat cagcctccat caaggttttg ttgaatatgg tatatgcttt
     2161 tccaaaccct gggacaagtt ttctcaggtg actgagacgt ctgaaactta ctgacttggt
     2221 ggtcatgatg gactctaatg cgtccagaca ttcctctctt ttcttgacta actcctccac
     2281 aacgagatgc tcaatctcgt ctgagcgaaa gtcgtgcaaa ttcaccaact gatctggagg
     2341 gcaccatttg gtctcatctg atgtttgcat cgcgacccat gttccatcca taagtctaag
     2401 tccaagaact ccacataact tgagcctgca tgctcctttt agagacttat acaggcctct
     2461 ttcatccaca aagccgcaag tcttgttccc gttggatgct ctcttccctc tgctattggt
     2521 aaaaatgtca caaggtgtcc ttggtctcgg attctcgggc atccaaatgg tgtaatcatg
     2581 gttagttgag cagtaggtag aggacaccgt tattcctgag cactttccgc cagggaagac
     2641 ccttgagtga agggatttgt catatgggtc caaatctgtc acacttgggg atatgataat
     2701 gagggactct ttggtggttc ttacagttcg aagccagtgg tagtcggggt atggattgtg
     2761 tagggactct tcatatctgg ggtcaccggc catcttccag ttatacgcgg ctctacatgc
     2821 gtctggggtg gggcggaaat gctttctctt gaatgtggtt gtgacataac caacaaagtt
     2881 ggtgtaggtc tctgcctctg tcacaacacc tgtgcaagtg aacccgttca ctttgatggc
     2941 tgagatgtat cccactttga gttccatgta ggagaactcg gacaggttgg tacatccttc
     3001 atcctccaca accaggttat ttggacagct gagatggtgt atgtcaatag ggctccaggg
     3061 accaagttcg tctggtatcg tgtaaatggg gaacttcccg aaacacaacg aaaaacccag
     3121 aaggagtaca aacaaaagaa cctgaggaac cataggtcca gggttctcct ccacgtctcc
     3181 agcctgcttc agcaggctga agttagtagc tccgcttcca gcgtaatctg gaacgtcata
     3241 tggataggat cctgcatagt ccgggacgtc atagggatag cccgcatagt caggaacatc
     3301 gtatgggtaa acggcgccgg atccatcctt agagctggtg tacttggtga tggccttagt
     3361 accctcggac acggcgtgct tggccaactc cccaggcagc agcaggcgca cggccgtctg
     3421 gatctccctg gaggtgatgg tcgagcgctt gttgtaatgc gccaggcggg aagcctcacc
     3481 tgcgatgcgc tcgaaaatgt cgttcacaaa cgagttcatg atgcccatgg ccttggacga
     3541 aatgccggtg tcagggtgga cctgcttcag aaccttgtac acatagatgg aatagctctc
     3601 cttgcggctg cgcttgcgct tcttgccgcc tttcttctgc gccttagtca ccgccttctt
     3661 ggagcccttt ttcggggcgg gagcagactt cgctggctct ggcatggtgg cggctctagc
     3721 ataacttcgt ataaagtatc ctatacgaag ttatttgcct taacccagaa attatcactg
     3781 ttattcttta gaatggtgca aagaataact tcgtataatg tatgctatac gaagttatga
     3841 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     3901 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3961 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     4021 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     4081 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     4141 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4201 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     4261 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt
     4321 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4381 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4441 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac
     4501 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     4561 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     4621 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     4681 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     4741 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     4801 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     4861 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     4921 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     4981 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     5041 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     5101 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     5161 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     5221 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     5281 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5341 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     5401 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     5461 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     5521 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     5581 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5641 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     5701 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     5761 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     5821 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     5881 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     5941 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     6001 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     6061 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     6121 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     6181 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     6241 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     6301 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     6361 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     6421 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     6481 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6541 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6601 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6661 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6721 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6781 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6841 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6901 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6961 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7021 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7081 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     7141 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7201 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     7261 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7321 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7381 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     7441 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7501 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7561 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7621 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7681 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
