LOCUS       sequence-193883-        7908 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 7908)
  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 7908)
  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-1EGFP
FEATURES             Location/Qualifiers
     source          1..7908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        59..438
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        439..642
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     593..613
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     687..706
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        687..705
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      854..863
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(860..862,863..865,866..1579)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(860..862,863..865,866..1576)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(905..926)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1166..1185)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1513..1534
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4983..5001)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4997..5044
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5067..5083)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5067..5083)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5120..5140)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5120..5138)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5159..5175)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5159..5175)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5172..5194)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5183..5199
                     /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(5207..5213,5214..5231,5232..5237))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5252..5273
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5390..5407)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5561..6149)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5641..5660)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6320..7111,7112..7180))
                     /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     6943..6962
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7181..7285)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7312..7768
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7399..7418)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7609..7630
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(7894..7908,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(7908,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgag cgcgcgataa gcttgatatc gaattcctgc agcgcgttga
       61 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      121 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      181 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      241 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      301 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      361 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      421 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      481 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      541 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      601 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa
      661 cccactgctt actggcttat cgaaattaat acgactcact atagggagac ccaagctggc
      721 tagattaagc gtctgatgag tccgtgagga cgaaacccgg cgtaccgggt cacgcttaac
      781 aaccagatca aagaaaaaac agacattgtc aattgcaaag caaaaatgta acacccctac
      841 accggtgaaa gccgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
      901 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
      961 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     1021 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     1081 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1141 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     1201 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1261 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     1321 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     1381 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1441 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     1501 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1561 ggacgagctg tacaagtaat gacctaggga agttgaataa caaaatgccg gaaatctacg
     1621 gattgtgtat atccatcatg aaaaaaacta acacccctcc tttcgaacca tcccaaacat
     1681 ggatgccgac aagattgtat tcaaagtcaa taatcaggtg gtctctttga agcctgagat
     1741 tatcgtggat caatatgagt acaagtaccc tgccatcaaa gatttgaaaa agccctgtat
     1801 aactctagga aaggctcccg atttaaataa agcatacaag tcagttttgt caggcatgag
     1861 cgccgccaaa cttaatcctg acgatgtatg ttcctatttg gcagcggcaa tgcagttttt
     1921 tgaggggaca tgtccggaag actggaccag ctatggaatt gtgattgcac gaaaaggaga
     1981 taagatcacc ccaggttctc tggtggagat aaaacgtact gatgtagaag ggaattgggc
     2041 tctgacagga ggcatggaac tgacaagaga ccccactgtc cctgagcatg cgtccttagt
     2101 cggtcttctc ttgagtctgt ataggttgag caaaatatcc gggcaaaaca ctggtaacta
     2161 taagacaaac attgcagaca ggatagagca gatttttgag acagcccctt ttgttaaaat
     2221 cgtggaacac catactctaa tgacaactca caaaatgtgt gctaattgga gtactatacc
     2281 aaacttcaga tttttggccg gaacctatga catgtttttc tcccggattg agcatctata
     2341 ttcagcaatc agagtgggca cagttgtcac tgcttatgaa gactgttcag gactggtatc
     2401 atttactggg ttcataaaac aaatcaatct caccgctaga gaggcaatac tatatttctt
     2461 ccacaagaac tttgaggaag agataagaag aatgtttgag ccagggcagg agacagctgt
     2521 tcctcactct tatttcatcc acttccgttc actaggcttg agtgggaaat ctccttattc
     2581 atcaaatgct gttggtcacg tgttcaatct cattcacttt gtaggatgct atatgggtca
     2641 agtcagatcc ctaaatgcaa cggttattgc tgcatgtgct cctcatgaaa tgtctgttct
     2701 agggggctat ctgggagagg aattcttcgg gaaagggaca tttgaaagaa gattcttcag
     2761 agatgagaaa gaacttcaag aatacgaggc ggctgaactg acaaagactg acgtagcact
     2821 ggcagatgat ggaactgtca actctgacga cgaggactac ttttcaggtg aaaccagaag
     2881 tccggaggct gtttatactc gaatcatgat gaatggaggt cgactaaaga gatctcacat
     2941 acggagatat gtctcagtca gttccaatca tcaagcccgt ccaaactcat tcgccgagtt
     3001 tctaaacaag acatattcga gtgactcata agaagttgaa taacaaaatg ccggaaatct
     3061 acggattgtg tatatccatc atgaaaaaaa ctaacacccc tcctttcgaa ccatcccaaa
     3121 catgagcaag atctttgtca atcctagtgc tattagagcc ggtctggccg atcttgagat
     3181 ggctgaagaa actgttgatc tgatcaatag aaatatcgaa gacaatcagg ctcatctcca
     3241 aggggaaccc atagaggtgg acaatctccc tgaggatatg gggcgacttc acctggatga
     3301 tggaaaatcg cccaaccatg gtgagatagc caaggtggga gaaggcaagt atcgagagga
     3361 ctttcagatg gatgaaggag aggatcctag cttcctgttc cagtcatacc tggaaaatgt
     3421 tggagtccaa atagtcagac aaatgaggtc aggagagaga tttctcaaga tatggtcaca
     3481 gaccgtagaa gagattatat cctatgtcgc ggtcaacttt cccaaccctc caggaaagtc
     3541 ttcagaggat aaatcaaccc agactactgg ccgagagctc aagaaggaga caacacccac
     3601 tccttctcag agagaaagcc aatcatcgaa agccaggatg gcggctcaaa ttgcttctgg
     3661 ccctccagcc cttgaatggt cggctaccaa tgaagaggat gatctatcag tggaggctga
     3721 gatcgctcac cagattgcag aaagtttctc caaaaaatat aagtttccct ctcgatcctc
     3781 agggatactc ttgtataatt ttgagcaatt gaaaatgaac cttgatgata tagttaaaga
     3841 ggcaaaaaat gtaccaggtg tgacccgttt agcccatgac gggtccaaac tccccctaag
     3901 atgtgtactg ggatgggtcg ctttggccaa ctctaagaaa ttccagttgt tagtcgaatc
     3961 cgacaagctg agtaaaatca tgcaagatga cttgaatcgc tatacatctt gctaaccgaa
     4021 cctctcccct cagtccctct agacaataaa atccgagatg tcccaaagtc aacatgaaaa
     4081 aaacaggcaa caccactgat aaaatgaacc tcctacgtaa gatagtgaaa aaccgcaggg
     4141 acgaggacac tcaaaaatcc tctcccgcgt cagcccctct ggatgacgat gacttgtggc
     4201 ttccaccccc tgaatacgtc ccgctgaaag aacttacagg caagaagaac atgaggaact
     4261 tttgtatcaa cggaagggtt aaagtgtgta gcccgaatgg ttactcgttc aggatcctgc
     4321 ggcacattct gaaatcattc gacgagatat attctgggaa tcataggatg atcgggttag
     4381 tcaaagtggt tattggactg gctttgtcag gatctccagt ccctgagggc ctgaactggg
     4441 tatacaaatt gaggagaacc tttatcttcc agtgggctga ttccaggggc cctcttgaag
     4501 gggaggagtt ggaatactct caggagatca cttgggatga tgatactgag ttcgtcggat
     4561 tgcaaataag agtgattgca aaacagtgtc atatccaggg cagagtctgg tgtatcaaca
     4621 tgaacccgag agcatgtcaa ctatggtctg acatgtctct tcagacacaa aggtccgaag
     4681 aggacaaaga ttcctctctg cttctagaat aaaccggata ctcctggaag cctgcccatg
     4741 ctaagactct tgtgtgatgt atcttgaaaa aaacaagatc ctaaatctga acctttggtt
     4801 gtttgattgt ttttctcatt tttgttgttt atttgttaag cgtgggtcgg catggcatct
     4861 ccacctcctc gcggtccgac ctgggcatcc gaaggaggac gcacgtccac tcggatggct
     4921 aagggagggc ggggatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     4981 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     5041 tttgctgaaa ggaggaacta tatccggatc cactagttct agagcggccg ccaccgcggt
     5101 ggagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     5161 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5221 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5281 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5341 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5401 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5461 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5521 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5581 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5641 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5701 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5761 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5821 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5881 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5941 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6001 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6061 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6121 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6181 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6241 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6301 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6361 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6421 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccaccc ttggaggctc
     6481 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6541 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6601 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6661 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6721 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6781 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6841 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6901 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6961 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7021 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7081 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7141 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7201 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7261 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct
     7321 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     7381 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     7441 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     7501 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     7561 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     7621 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     7681 tgccgatttc ggcctattgg ttaaaaaaat gagctgattt aacaaaaatt taacgcgaat
     7741 tttaacaaaa tattaacgct tacaatttcc attcgccatt caggctgcgc aactgttggg
     7801 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     7861 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgtt
//
