LOCUS       sequence-485417-        7802 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 7802)
  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 7802)
  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: pAM-CAGGS-FLEX-H2B-3XHA-P2A-N2c(G)
FEATURES             Location/Qualifiers
     source          1..7802
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   34..163
                     /label=AAV2 ITR
     promoter        469..746
                     /label=chicken beta-actin promoter
     primer_bind     1025..1047
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1109..1128
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1163..1196
                     /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    1259..1292
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2900..2956)
                     /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(2966..3055)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             complement(3074..3451)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSSK"
     protein_bind    complement(3479..3512)
                     /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(3583..3616)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3655..4243
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3708..3728)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4126..4137)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4270..4287)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4276..4500
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(4710..5039)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4724..4859
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5024..5044
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(5285..5873)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5365..5384)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6044..6835,6836..6904))
                     /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     6667..6686
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6905..7009)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7077..7095
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7133..7155)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7255..7274
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7488..7507
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(7797..7802,1..13)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(7797..7802,1..12)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gtgacactat agaatacacg gaattaattc tagctgcgcg ctcgctcgct cactgaggcc
       61 gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga
      121 gcgcgcagag agggagtggc caactccatc actaggggtt ccttgtagtt aatgattaac
      181 ccgccatgct acttatctac gtagccatgc tctaggtacc attgacgtca ataatgacgt
      241 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      301 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      361 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      421 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc
      481 ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat
      541 ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggg sgcgcgccag
      601 gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca
      661 atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct
      721 ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc
      781 gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg
      841 tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc
      901 ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg
      961 gggggagcgg ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg
     1021 gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat
     1081 gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat
     1141 tttggcaaag aattggatcc ccataacttc gtataaagta tcctatacga agttatatca
     1201 aaataggaag accaatgctt caccatcgac ccgaattgcc aagcatcacc atcgacccat
     1261 aacttcgtat aatgtatgct atacgaagtt atgaattcct gcaggagctc actagtgcgg
     1321 ccgcttacag tctgatctca cctccactct tatatgattc ccatgaaggt atgacttttc
     1381 cgctttggga agtgactgac acattccccc ctgtccctcc aaaactgcgt tgtttcgatt
     1441 ctggtcgatt ggctcttctg caccatgtca ttagggaaaa tatcaacacc aggccaatca
     1501 tggcccctgc agtcatcaat acatactttc cccagttcgg gagacccagg tcaacccctg
     1561 agatctgttt gtacacatcg gggaggtgaa cttcaacaaa atcctcagcc tcatcacctt
     1621 ctttgaaaac tgtagaaggg tctgccaggg ggtgcatcag ggggataact gaagattcca
     1681 acaactccat atgttgctgg aggagggatg attgcatctc tgggattagg acatggtcgt
     1741 caggccctaa tattatacca ttgaaaaaca ccccgttcac atgaggatgg caccttcctc
     1801 caactttcaa acaccctttt gaggggatga tctcattcca ggtccggact gacttgtagt
     1861 gagcatcagc ctccatcaag gttttgttga atatggtata tgcttttcca aaccctggga
     1921 caagttttct caggtgactg agacgtctga aacttactga cttggtggtc atgatggact
     1981 ctaatgcgtc cagacattcc tctcttttct tgactaactc ctccacaacg agatgctcaa
     2041 tctcgtctga gcgaaagtcg tgcaaattca ccaactgatc tggagggcac catttggtct
     2101 catctgatgt ttgcatcgcg acccatgttc catccataag tctaagtcca agaactccac
     2161 ataacttgag cctgcatgct ccttttagag acttatacag gcctctttca tccacaaagc
     2221 cgcaagtctt gttcccgttg gatgctctct tccctctgct attggtaaaa atgtcacaag
     2281 gtgtccttgg tctcggattc tcgggcatcc aaatggtgta atcatggtta gttgagcagt
     2341 aggtagagga caccgttatt cctgagcact ttccgccagg gaagaccctt gagtgaaggg
     2401 atttgtcata tgggtccaaa tctgtcacac ttggggatat gataatgagg gactctttgg
     2461 tggttcttac agttcgaagc cagtggtagt cggggtatgg attgtgtagg gactcttcat
     2521 atctggggtc accggccatc ttccagttat acgcggctct acatgcgtct ggggtggggc
     2581 ggaaatgctt tctcttgaat gtggttgtga cataaccaac aaagttggtg taggtctctg
     2641 cctctgtcac aacacctgtg caagtgaacc cgttcacttt gatggctgag atgtatccca
     2701 ctttgagttc catgtaggag aactcggaca ggttggtaca tccttcatcc tccacaacca
     2761 ggttatttgg acagctgaga tggtgtatgt caatagggct ccagggacca agttcgtctg
     2821 gtatcgtgta aatggggaac ttcccgaaac acaacgaaaa acccagaagg agtacaaaca
     2881 aaagaacctg aggaaccata ggtccagggt tctcctccac gtctccagcc tgcttcagca
     2941 ggctgaagtt agtagctccg cttccagcgt aatctggaac gtcatatgga taggatcctg
     3001 catagtccgg gacgtcatag ggatagcccg catagtcagg aacatcgtat gggtaaacgg
     3061 cgccggatcc atccttagag ctggtgtact tggtgatggc cttagtaccc tcggacacgg
     3121 cgtgcttggc caactcccca ggcagcagca ggcgcacggc cgtctggatc tccctggagg
     3181 tgatggtcga gcgcttgttg taatgcgcca ggcgggaagc ctcacctgcg atgcgctcga
     3241 aaatgtcgtt cacaaacgag ttcatgatgc ccatggcctt ggacgaaatg ccggtgtcag
     3301 ggtggacctg cttcagaacc ttgtacacat agatggaata gctctccttg cggctgcgct
     3361 tgcgcttctt gccgcctttc ttctgcgcct tagtcaccgc cttcttggag ccctttttcg
     3421 gggcgggagc agacttcgct ggctctggca tggtggcggc ctcgagagat ctgatatcat
     3481 aacttcgtat aggatacttt atacgaagtt atcattggga ttcttcctat tttgatccaa
     3541 gcatcaccat cgaccctcta gtccagatct caccatcgac ccataacttc gtatagcata
     3601 cattatacga agttatgtcc ctcgaagagg ttcaagcttt aacagcttat cgataatcaa
     3661 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3721 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3781 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgaata gttgtggccc
     3841 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     3901 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     3961 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4021 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     4081 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     4141 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     4201 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgactc
     4261 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     4321 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     4381 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     4441 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     4501 cttctgaggc ggaaagaacc agctggggct cgactagagc atggctacgt agataagtag
     4561 catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct
     4621 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcggc
     4681 ctcagtgagc gagcgagcgc gcagagcttt ttgcaaaagc ctaggcctcc aaaaaagcct
     4741 cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa
     4801 aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg
     4861 ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt
     4921 ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg
     4981 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca
     5041 ttccacagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5101 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5161 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5221 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5281 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5341 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5401 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5461 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5521 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5581 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5641 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5701 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5761 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5821 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5881 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5941 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6001 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6061 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6121 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6181 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6241 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6301 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6361 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6421 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6481 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6541 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6601 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6661 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6721 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6781 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6841 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6901 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6961 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7021 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     7081 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     7141 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     7201 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     7261 cagagcagat tgtactgaga gtgcaccatt cgacgctctc ccttatgcga ctcctgcatt
     7321 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     7381 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     7441 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     7501 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     7561 tagaggatct ggctagcgat gaccctgctg attggttcgc tgaccatttc cgggtgcggg
     7621 acggcgttac cagaaactca gaaggttcgt ccaaccaaac cgactctgac ggcagtttac
     7681 gagagagatg atagggtctg cttcagtaag ccagatgcta cacaattagg cttgtacata
     7741 ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca catacgattt
     7801 ag
//
