LOCUS       sequence-193881-        8593 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Second-generation rabies viral vector genome.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8593)
  AUTHORS   Chatterjee S, Sullivan HA, MacLennan BJ, Xu R, Hou Y, Lavin TK, Lea 
            NE, Michalski JE, Babcock KR, Dietrich S, Matthews GA, Beyeler A, 
            Calhoon GG, Glober G, Whitesell JD, Yao S, Cetin A, Harris JA, Zeng 
            H, Tye KM, Reid RC, Wickersham IR
  TITLE     Nontoxic, double-deletion-mutant rabies viral vectors for retrograde
            targeting of projection neurons.
  JOURNAL   Nat Neurosci. 2018 Apr;21(4):638-646. doi: 
            10.1038/s41593-018-0091-7. Epub 2018 Mar 5.
  PUBMED    29507411
REFERENCE   2  (bases 1 to 8593)
  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"; doi: 
            "10.1038/s41593-018-0091-7"; journalName: "Nat Neurosci"; date: 
            "2018-04"; volume: "21"; issue: "4"; pages: "638-646"
COMMENT     Sequence Label: pRVdGL-4Flpo
FEATURES             Location/Qualifiers
     source          1..8593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(533..1324,1325..1393))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGASKGGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1156..1175
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1394..1498)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1525..1981
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1612..1631)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1822..1843
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2107..2129
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2121..2138
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2122..2138
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        2180..2559
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2560..2763
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2714..2734
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2808..2827
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2808..2826
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      6211..6220
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6217..6222,6223..6243,6244..7515)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     primer_bind     complement(7789..7807)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7803..7850
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(7873..7889)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7873..7889)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(7926..7946)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7926..7944)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7965..7981)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7965..7981)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7978..8000)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7989..8005
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(8013..8019,8020..8037,8038..8043))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8058..8079
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8196..8213)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(8367..8593,1..362))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8447..8466)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
       61 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      121 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      181 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
      241 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      301 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
      361 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
      421 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
      481 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
      541 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
      601 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
      661 caatgatacc gcgagaccca cccttggagg ctccagattt atcagcaata aaccagccag
      721 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
      781 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
      841 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
      901 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
      961 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     1021 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1081 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     1141 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     1201 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     1261 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     1321 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     1381 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     1441 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     1501 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     1561 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     1621 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     1681 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     1741 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     1801 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     1861 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     1921 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     1981 tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     2041 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     2101 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgata agcttgatat
     2161 cgaattcctg cagcgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     2221 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     2281 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     2341 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     2401 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     2461 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     2521 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     2581 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     2641 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     2701 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     2761 gctctctggc taactagaga acccactgct tactggctta tcgaaattaa tacgactcac
     2821 tatagggaga cccaagctgg ctagattaag cgtctgatga gtccgtgagg acgaaacccg
     2881 gcgtaccggg tcacgcttaa caaccagatc aaagaaaaaa cagacattgt caattgcaaa
     2941 gcaaaaatgt aacaccccta caatggatgc cgacaagatt gtattcaaag tcaataatca
     3001 ggtggtctct ttgaagcctg agattatcgt ggatcaatat gagtacaagt accctgccat
     3061 caaagatttg aaaaagccct gtataaccct aggaaaggct cccgatttaa ataaagcata
     3121 caagtcagtt ttgtcaggca tgagcgccgc caaacttaat cctgacgatg tatgttccta
     3181 tttggcagcg gcaatgcagt tttttgaggg gacatgtccg gaagactgga ccagctatgg
     3241 aattgtgatt gcacgaaaag gagataagat caccccaggt tctctggtgg agataaaacg
     3301 tactgatgta gaagggaatt gggctctgac aggaggcatg gaactgacaa gagaccccac
     3361 tgtccctgag catgcgtcct tagtcggtct tctcttgagt ctgtataggt tgagcaaaat
     3421 atccgggcaa aacactggta actataagac aaacattgca gacaggatag agcagatttt
     3481 tgagacagcc ccttttgtta aaatcgtgga acaccatact ctaatgacaa ctcacaaaat
     3541 gtgtgctaat tggagtacta taccaaactt cagatttttg gccggaacct atgacatgtt
     3601 tttctcccgg attgagcatc tatattcagc aatcagagtg ggcacagttg tcactgctta
     3661 tgaagactgt tcaggactgg tatcatttac tgggttcata aaacaaatca atctcaccgc
     3721 tagagaggca atactatatt tcttccacaa gaactttgag gaagagataa gaagaatgtt
     3781 tgagccaggg caggagacag ctgttcctca ctcttatttc atccacttcc gttcactagg
     3841 cttgagtggg aaatctcctt attcatcaaa tgctgttggt cacgtgttca atctcattca
     3901 ctttgtagga tgctatatgg gtcaagtcag atccctaaat gcaacggtta ttgctgcatg
     3961 tgctcctcat gaaatgtctg ttctaggggg ctatctggga gaggaattct tcgggaaagg
     4021 gacatttgaa agaagattct tcagagatga gaaagaactt caagaatacg aggcggctga
     4081 actgacaaag actgacgtag cactggcaga tgatggaact gtcaactctg acgacgagga
     4141 ctacttttca ggtgaaacca gaagtccgga ggctgtttat actcgaatca tgatgaatgg
     4201 aggtcgacta aagagatctc acatacggag atatgtctca gtcagttcca atcatcaagc
     4261 ccgtccaaac tcattcgccg agtttctaaa caagacatat tcgagtgact cataagaagt
     4321 tgaataacaa aatgccggaa atctacggat tgtgtatatc catcatgaaa aaaactaaca
     4381 cccctccttt cgaaccatcc caaacatgag caagatcttt gtcaatccta gtgctattag
     4441 agccggtctg gccgatcttg agatggctga agaaactgtt gatctgatca atagaaatat
     4501 cgaagacaat caggctcatc tccaagggga acccatagag gtggacaatc tccctgagga
     4561 tatggggcga cttcacctgg atgatggaaa atcgcccaac catggtgaga tagccaaggt
     4621 gggagaaggc aagtatcgag aggactttca gatggatgaa ggagaggatc ctagcttcct
     4681 gttccagtca tacctggaaa atgttggagt ccaaatagtc agacaaatga ggtcaggaga
     4741 gagatttctc aagatatggt cacagaccgt agaagagatt atatcctatg tcgcggtcaa
     4801 ctttcccaac cctccaggaa agtcttcaga ggataaatca acccagacta ctggccgaga
     4861 gctcaagaag gagacaacac ccactccttc tcagagagaa agccaatcat cgaaagccag
     4921 gatggcggct caaattgctt ctggccctcc agcccttgaa tggtcggcta ccaatgaaga
     4981 ggatgatcta tcagtggagg ctgagatcgc tcaccagatt gcagaaagtt tctccaaaaa
     5041 atataagttt ccctctcgat cctcagggat actcttgtat aattttgagc aattgaaaat
     5101 gaaccttgat gatatagtta aagaggcaaa aaatgtacca ggtgtgaccc gtttagccca
     5161 tgacgggtcc aaactccccc taagatgtgt actgggatgg gtcgctttgg ccaactctaa
     5221 gaaattccag ttgttagtcg aatccgacaa gctgagtaaa atcatgcaag atgacttgaa
     5281 tcgctataca tcttgctaac cgaacctctc ccctcagtcc ctctagacaa taaaatccga
     5341 gatgtcccaa agtcaacatg aaaaaaacag gcaacaccac tgataaaatg aacctcctac
     5401 gtaagatagt gaaaaaccgc agggacgagg acactcaaaa atcctctccc gcgtcagccc
     5461 ctctggatga cgatgacttg tggcttccac cccctgaata cgtcccgctg aaagaactta
     5521 caggcaagaa gaacatgagg aacttttgta tcaacggaag ggttaaagtg tgtagcccga
     5581 atggttactc gttcaggatc ctgcggcaca ttctgaaatc attcgacgag atatattctg
     5641 ggaatcatag gatgatcggg ttagtcaaag tggttattgg actggctttg tcaggatctc
     5701 cagtccctga gggcctgaac tgggtataca aattgaggag aacctttatc ttccagtggg
     5761 ctgattccag gggccctctt gaaggggagg agttggaata ctctcaggag atcacttggg
     5821 atgatgatac tgagttcgtc ggattgcaaa taagagtgat tgcaaaacag tgtcatatcc
     5881 agggcagagt ctggtgtatc aacatgaacc cgagagcatg tcaactatgg tctgacatgt
     5941 ctcttcagac acaaaggtcc gaagaggaca aagattcctc tctgcttcta gaataatcag
     6001 attatatccc gcaaatttat cacttgttta cctctggagg agagaacata tgggctcaac
     6061 tccaaccctt gggagcaata taacaaaaaa catgttatgg tgccattaaa ccgctgcatt
     6121 tcatcaaagt caagttgatt acctttacat tttgatcctc ttggatgtga aaaaaactat
     6181 taacatccct caaaagaccc cgggaaagcc gccaccatgg ctcctaagaa gaagaggaag
     6241 gtgatgagcc agttcgacat cctgtgcaag acccccccca aggtgctggt gcggcagttc
     6301 gtggagagat tcgagaggcc cagcggcgag aagatcgcca gctgtgccgc cgagctgacc
     6361 tacctgtgct ggatgatcac ccacaacggc accgccatca agagggccac cttcatgagc
     6421 tacaacacca tcatcagcaa cagcctgagc ttcgacatcg tgaacaagag cctgcagttc
     6481 aagtacaaga cccagaaggc caccatcctg gaggccagcc tgaagaagct gatccccgcc
     6541 tgggagttca ccatcatccc ttacaacggc cagaagcacc agagcgacat caccgacatc
     6601 gtgtccagcc tgcagctgca gttcgagagc agcgaggagg ccgacaaggg caacagccac
     6661 agcaagaaga tgctgaaggc cctgctgtcc gagggcgaga gcatctggga gatcaccgag
     6721 aagatcctga acagcttcga gtacaccagc aggttcacca agaccaagac cctgtaccag
     6781 ttcctgttcc tggccacatt catcaactgc ggcaggttca gcgacatcaa gaacgtggac
     6841 cccaagagct tcaagctggt gcagaacaag tacctgggcg tgatcattca gtgcctggtg
     6901 accgagacca agacaagcgt gtccaggcac atctactttt tcagcgccag aggcaggatc
     6961 gaccccctgg tgtacctgga cgagttcctg aggaacagcg agcccgtgct gaagagagtg
     7021 aacaggaccg gcaacagcag cagcaacaag caggagtacc agctgctgaa ggacaacctg
     7081 gtgcgcagct acaacaaggc cctgaagaag aacgccccct accccatctt cgctatcaag
     7141 aacggcccta agagccacat cggcaggcac ctgatgacca gctttctgag catgaagggc
     7201 ctgaccgagc tgacaaacgt ggtgggcaac tggagcgaca agagggcctc cgccgtggcc
     7261 aggaccacct acacccacca gatcaccgcc atccccgacc actacttcgc cctggtgtcc
     7321 aggtactacg cctacgaccc catcagcaag gagatgatcg ccctgaagga cgagaccaac
     7381 cccatcgagg agtggcagca catcgagcag ctgaagggca gcgccgaggg cagcatcaga
     7441 taccccgcct ggaacggcat catcagccag gaggtgctgg actacctgag cagctacatc
     7501 aacaggcgga tctgatgaac cggatactcc tggaagcctg cccatgctaa gactcttgtg
     7561 tgatgtatct tgaaaaaaac aagatcctaa atctgaacct ttggttgttt gattgttttt
     7621 ctcatttttg ttgtttattt gttaagcgtg ggtcggcatg gcatctccac ctcctcgcgg
     7681 tccgacctgg gcatccgaag gaggacgcac gtccactcgg atggctaagg gagggcgggg
     7741 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     7801 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     7861 gaactatatc cggatccact agttctagag cggccgccac cgcggtggag ctccagcttt
     7921 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     7981 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     8041 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     8101 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     8161 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     8221 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     8281 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     8341 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     8401 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     8461 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     8521 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     8581 ctcagttcgg tgt
//
