LOCUS       sequence-193890-        8014 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 8014)
  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 8014)
  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-4EGFP
FEATURES             Location/Qualifiers
     source          1..8014
                     /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      4090..4099
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4096..4098,4099..4101,4102..4815)
                     /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(4096..4098,4099..4101,4102..4812)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4141..4162)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4402..4421)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4749..4770
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5089..5107)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5103..5150
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5173..5189)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5173..5189)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5226..5246)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5226..5244)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5265..5281)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5265..5281)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5278..5300)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5289..5305
                     /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(5313..5319,5320..5337,5338..5343))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5358..5379
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5496..5513)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5667..6255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5747..5766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6426..7217,7218..7286))
                     /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     7049..7068
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7287..7391)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7418..7874
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7505..7524)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7715..7736
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(8000..8014,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(8014,1..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 aatggatgcc gacaagattg tattcaaagt caataatcag gtggtctctt tgaagcctga
      901 gattatcgtg gatcaatatg agtacaagta ccctgccatc aaagatttga aaaagccctg
      961 tataacccta ggaaaggctc ccgatttaaa taaagcatac aagtcagttt tgtcaggcat
     1021 gagcgccgcc aaacttaatc ctgacgatgt atgttcctat ttggcagcgg caatgcagtt
     1081 ttttgagggg acatgtccgg aagactggac cagctatgga attgtgattg cacgaaaagg
     1141 agataagatc accccaggtt ctctggtgga gataaaacgt actgatgtag aagggaattg
     1201 ggctctgaca ggaggcatgg aactgacaag agaccccact gtccctgagc atgcgtcctt
     1261 agtcggtctt ctcttgagtc tgtataggtt gagcaaaata tccgggcaaa acactggtaa
     1321 ctataagaca aacattgcag acaggataga gcagattttt gagacagccc cttttgttaa
     1381 aatcgtggaa caccatactc taatgacaac tcacaaaatg tgtgctaatt ggagtactat
     1441 accaaacttc agatttttgg ccggaaccta tgacatgttt ttctcccgga ttgagcatct
     1501 atattcagca atcagagtgg gcacagttgt cactgcttat gaagactgtt caggactggt
     1561 atcatttact gggttcataa aacaaatcaa tctcaccgct agagaggcaa tactatattt
     1621 cttccacaag aactttgagg aagagataag aagaatgttt gagccagggc aggagacagc
     1681 tgttcctcac tcttatttca tccacttccg ttcactaggc ttgagtggga aatctcctta
     1741 ttcatcaaat gctgttggtc acgtgttcaa tctcattcac tttgtaggat gctatatggg
     1801 tcaagtcaga tccctaaatg caacggttat tgctgcatgt gctcctcatg aaatgtctgt
     1861 tctagggggc tatctgggag aggaattctt cgggaaaggg acatttgaaa gaagattctt
     1921 cagagatgag aaagaacttc aagaatacga ggcggctgaa ctgacaaaga ctgacgtagc
     1981 actggcagat gatggaactg tcaactctga cgacgaggac tacttttcag gtgaaaccag
     2041 aagtccggag gctgtttata ctcgaatcat gatgaatgga ggtcgactaa agagatctca
     2101 catacggaga tatgtctcag tcagttccaa tcatcaagcc cgtccaaact cattcgccga
     2161 gtttctaaac aagacatatt cgagtgactc ataagaagtt gaataacaaa atgccggaaa
     2221 tctacggatt gtgtatatcc atcatgaaaa aaactaacac ccctcctttc gaaccatccc
     2281 aaacatgagc aagatctttg tcaatcctag tgctattaga gccggtctgg ccgatcttga
     2341 gatggctgaa gaaactgttg atctgatcaa tagaaatatc gaagacaatc aggctcatct
     2401 ccaaggggaa cccatagagg tggacaatct ccctgaggat atggggcgac ttcacctgga
     2461 tgatggaaaa tcgcccaacc atggtgagat agccaaggtg ggagaaggca agtatcgaga
     2521 ggactttcag atggatgaag gagaggatcc tagcttcctg ttccagtcat acctggaaaa
     2581 tgttggagtc caaatagtca gacaaatgag gtcaggagag agatttctca agatatggtc
     2641 acagaccgta gaagagatta tatcctatgt cgcggtcaac tttcccaacc ctccaggaaa
     2701 gtcttcagag gataaatcaa cccagactac tggccgagag ctcaagaagg agacaacacc
     2761 cactccttct cagagagaaa gccaatcatc gaaagccagg atggcggctc aaattgcttc
     2821 tggccctcca gcccttgaat ggtcggctac caatgaagag gatgatctat cagtggaggc
     2881 tgagatcgct caccagattg cagaaagttt ctccaaaaaa tataagtttc cctctcgatc
     2941 ctcagggata ctcttgtata attttgagca attgaaaatg aaccttgatg atatagttaa
     3001 agaggcaaaa aatgtaccag gtgtgacccg tttagcccat gacgggtcca aactccccct
     3061 aagatgtgta ctgggatggg tcgctttggc caactctaag aaattccagt tgttagtcga
     3121 atccgacaag ctgagtaaaa tcatgcaaga tgacttgaat cgctatacat cttgctaacc
     3181 gaacctctcc cctcagtccc tctagacaat aaaatccgag atgtcccaaa gtcaacatga
     3241 aaaaaacagg caacaccact gataaaatga acctcctacg taagatagtg aaaaaccgca
     3301 gggacgagga cactcaaaaa tcctctcccg cgtcagcccc tctggatgac gatgacttgt
     3361 ggcttccacc ccctgaatac gtcccgctga aagaacttac aggcaagaag aacatgagga
     3421 acttttgtat caacggaagg gttaaagtgt gtagcccgaa tggttactcg ttcaggatcc
     3481 tgcggcacat tctgaaatca ttcgacgaga tatattctgg gaatcatagg atgatcgggt
     3541 tagtcaaagt ggttattgga ctggctttgt caggatctcc agtccctgag ggcctgaact
     3601 gggtatacaa attgaggaga acctttatct tccagtgggc tgattccagg ggccctcttg
     3661 aaggggagga gttggaatac tctcaggaga tcacttggga tgatgatact gagttcgtcg
     3721 gattgcaaat aagagtgatt gcaaaacagt gtcatatcca gggcagagtc tggtgtatca
     3781 acatgaaccc gagagcatgt caactatggt ctgacatgtc tcttcagaca caaaggtccg
     3841 aagaggacaa agattcctct ctgcttctag aataatcaga ttatatcccg caaatttatc
     3901 acttgtttac ctctggagga gagaacatat gggctcaact ccaacccttg ggagcaatat
     3961 aacaaaaaac atgttatggt gccattaaac cgctgcattt catcaaagtc aagttgatta
     4021 cctttacatt ttgatcctct tggatgtgaa aaaaactatt aacatccctc aaaagacccc
     4081 gggaaagccg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     4141 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     4201 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     4261 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     4321 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     4381 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     4441 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     4501 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     4561 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     4621 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     4681 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     4741 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     4801 gagctgtaca agtaatgaac cggatactcc tggaagcctg cccatgctaa gactcttgtg
     4861 tgatgtatct tgaaaaaaac aagatcctaa atctgaacct ttggttgttt gattgttttt
     4921 ctcatttttg ttgtttattt gttaagcgtg ggtcggcatg gcatctccac ctcctcgcgg
     4981 tccgacctgg gcatccgaag gaggacgcac gtccactcgg atggctaagg gagggcgggg
     5041 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     5101 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     5161 gaactatatc cggatccact agttctagag cggccgccac cgcggtggag ctccagcttt
     5221 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     5281 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5341 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5401 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5461 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5521 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5581 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5641 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5701 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5761 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5821 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5881 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5941 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6001 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6061 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6121 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6181 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6241 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6301 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6361 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6421 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6481 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6541 ggccccagtg ctgcaatgat accgcgagac ccacccttgg aggctccaga tttatcagca
     6601 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6661 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6721 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6781 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6841 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6901 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6961 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7021 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7081 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7141 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7201 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7261 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7321 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7381 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta
     7441 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     7501 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     7561 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     7621 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     7681 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     7741 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     7801 tattggttaa aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     7861 aacgcttaca atttccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     7921 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     7981 tgggtaacgc cagggttttc ccagtcacga cgtt
//
