LOCUS       sequence-463642-        9575 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral helper virus for G-deleted rabies tracing. 
            mGreenLantern:KASH fluorescence is not visible without 
            immunostaining..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9575)
  AUTHORS   Shin D, Urbanek ME, Larson HH, Moussa AJ, Lee KY, Baker DL, 
            Standen-Bloom E, Ramachandran S, Bogdanoff D, Cadwell CR, Nowakowski
            TJ
  TITLE     High-Complexity Barcoded Rabies Virus for Scalable Circuit Mapping 
            Using Single-Cell and Single-Nucleus Sequencing.
  JOURNAL   bioRxiv [Preprint]. 2024 Dec 11:2024.10.01.616167. doi: 
            10.1101/2024.10.01.616167.
  PUBMED    39713304
REFERENCE   2  (bases 1 to 9575)
  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"; journalName: "bioRxiv 
            [Preprint]. 2024 Dec 11:2024.10.01.616167. doi: 
            10.1101/2024.10.01.616167."
COMMENT     Sequence Label: pSico-syn-mGreenLantern:KASH-TVA-oG-6xMBS-WPRE...
FEATURES             Location/Qualifiers
     source          1..9575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     523..542
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        617..721
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(722..790,791..1582)
                     /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(940..959)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1753..2341
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2242..2261
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             2403..2583
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        2976..3423
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      3437..3446
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4433..4489
                     /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             4961..5014
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     complement(6691..6711)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    7321..7545
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    7603..7720
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(8018..8062)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    8247..8480
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    8977..9102
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             9149..9329
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(9347..9545)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(9372..9392)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(9547..9575,1..351)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
       61 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
      121 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
      181 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
      241 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
      301 ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt caacgcgtat
      361 atctggcccg tacatcgcga agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc
      421 aattgtcggt caagccttgc cttgttgtag cttaaatttt gctcgcgcac tactcagcga
      481 cctccaacac acaagcaggg agcagatact ggcttaacta tgcggcatca gagcagattg
      541 tactgagagt gcaccatagg ggatcgggag atctcccgat ccgtcgacgt caggtggcac
      601 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
      661 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
      721 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
      781 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
      841 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
      901 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
      961 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     1021 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     1081 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     1141 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     1201 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     1261 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     1321 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     1381 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     1441 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     1501 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     1561 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     1621 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     1681 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     1741 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     1801 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     1861 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     1921 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     1981 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2041 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2101 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2161 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2221 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2281 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2341 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2401 gctgctagag attttccaca ctgactaaaa gggtctgagg gatctctagt taccagagtc
     2461 acacaacaga cgggcacaca ctacttgaag cactcaaggc aagctttatt gaggcttaag
     2521 cagtgggttc cctagttagc cagagagctc ccaggctcag atctggtcta accagagaga
     2581 cccagtacag tccggatgca gctctcgggc catgtgatga aatgctaggc ggctgtcaaa
     2641 cctccactct aatacttctc tctccgggtc atccatccca tgcaggctca cagggtgtaa
     2701 caagcgggtg ttctctcctt cattggcttc ttctaccttc tcttgctcaa ctggtactag
     2761 cttgtagcac catccaaagg tcagtggata tctgatccct ggccctggtg tgtagttctg
     2821 ccaatcaggg aagtagcctt gtgtgtggta gatccacaga tcaaggatat cttgtcttcg
     2881 ttgggagtga attagccctt ccagtccccc cttttctttt aaaaagtggc taagatctac
     2941 agctgccttg taagtcattg gtcttaaagg tcaccagtgc aagtgggttt taggaccagg
     3001 atgaggcggg gtgggggtgc ctacctgacg accgaccccg acccactgga caagcaccca
     3061 acccccattc cccaaattgc gcatccccta tcagagaggg ggaggggaaa caggatgcgg
     3121 cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag tgccttcgcc cccgcctggc
     3181 ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga cgtcactcgc cggtcccccg
     3241 caaactcccc ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc agccggaccg
     3301 caccacgcga ggcgcgagat aggggggcac gggcgcgacc atctgcgctg cggcgccggc
     3361 gactcagcgc tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg
     3421 cagtcgagaa accggtgcca ccatggtgtc aaaaggtggg gagctcttta caggagtagt
     3481 gccgattctc gtcgagttgg atggtgacgt gaacggccat aagttctcag tccgagggga
     3541 aggggaggga gatgctacta acggtaaact cactctcaag ttcatatgta ctacaggtaa
     3601 actcccggtg ccctggccaa cccttgtaac tactttgggg tacggagtcg cctgtttcgc
     3661 gaggtatccc gatcacatga agcagcacga cttctttaag agcgctatgc ctgagggata
     3721 tgtccaggag cgaactatct ctttcaaaga cgatgggaca tacaagactc gagccgaagt
     3781 gaaatttgaa ggggatactc tcgttaatcg gatcgtattg aagggaatag atttcaagga
     3841 ggatggtaat atattgggac ataagttgga atataacttt aactcccata aagtttacat
     3901 cactgcggat aagcagaaaa acggcattaa ggccaatttc aagactcgcc acaatgtaga
     3961 ggatggaggc gtacaacttg ctgaccatta tcagcagaat accccaatag gagacgggcc
     4021 ggtgttgctc cccgataacc actacttgtc acaccagtct aagctgtcca aagatccgaa
     4081 cgaaaaaagg gatcacatgg ttcttaaaga aagggtaacc gccgccggaa tcacgcacga
     4141 catggatgaa ctgtacaagt ccggtttgag gtcaagagaa gaagaggagg agactgactc
     4201 ccgaatgccg cacttggact caccagggag tagtcagcct agacgcagtt ttctgagccg
     4261 cgtgataaga gccgccctgc cccttcagtt gctgcttctc ctgcttctct tgctcgcgtg
     4321 tctgctcccc gcatctgaag atgattactc ctgtacccag gctaacaact tcgcacgatc
     4381 attttatcct atgttgagat ataccaacgg accccctcct acaggttcgg gtgctacaaa
     4441 tttttctctc ctgaagcagg cgggtgacgt tgaagagaat ccaggaccta tggcgcgcct
     4501 gttgcctgcg cttctgctgc ttttgttgcc tggtaatgta accggaaatg gctcaggaaa
     4561 cgggtcactc tctcgatgcc ctcccggtca gttccgatgt agcgagcctc ccggggcaca
     4621 cggcgagtgc tacccacaag actggttgtg tgacgggcat cctgattgtg acgacggtcg
     4681 agatgaatgg ggatgtggta ctagtgcgat cccagcagtg cctacagata atggcacgga
     4741 agccccgaca gctccggcgc ctggacgggc tctcccagca cgcaaccacg gacgcatgtg
     4801 gatgctcatt actgccgttc tcctgtgctg tctcgtagct gtgggtggga ttgcagcttg
     4861 ggggaagtct aaggcgaaaa gcaggagtga catctttagt ctggaatcag ctagtaagga
     4921 gctcctcgtc ccagacaagt cccaagcgga tcttttttcc gaagggcgcg gtagccttct
     4981 gacgtgcgga gatgtggagg aaaatcctgg acctgcaggc accatggtgc cccaagcgct
     5041 gttgttcgtt cctctgttgg tgttcccact gtgctttggt aaattcccta tttacaccat
     5101 accggataag ctcggtccgt ggtctcctat cgacatccac catttgtctt gccctaacaa
     5161 cctggtagta gaagatgagg gatgcaccaa cttgagtggg ttctcctata tggagctcaa
     5221 ggtagggtat atttcagcca tcaaaatgaa tggttttacg tgtacaggtg ttgtaactga
     5281 agctgagaca tacactaact ttgtcgggta tgtaaccacc acgtttaaaa gaaaacattt
     5341 tagaccaacg cctgacgcgt gccgagcagc ttacaattgg aaaatggctg gcgatccgag
     5401 gtatgaagag tcactgcata atccctaccc agattaccat tggttgcgaa ctgtgaaaac
     5461 gacaaaagaa tccctcgtaa tcattagtcc ctcagtcgca gatctggacc cttacgaccg
     5521 gagcctgcac agtcctgtgt tcccgggcgg caattgtagc ggcgtagcgg tgagttccac
     5581 ctattgcagc accaaccatg attacacgat ctggatgccg gagaatccca ggttgggtat
     5641 gagttgtgac atattcacta actctcgcgg caagcgggcg tccaagggta gtgagacgtg
     5701 tggatttgta gatgaaaggg gcctttacaa atcattgaaa ggtgcctgca aactcaaact
     5761 ctgcggcgtc cttggtcttc ggttgatgga tggaacctgg gtggcgatgc aaacttccaa
     5821 cgagacaaag tggtgtccac caggccagct ggtgaatctc cacgacttcc ggagtgatga
     5881 aatagaacat ttggtcgttg aagaacttgt gaagaagaga gaagagtgcc tggacgctct
     5941 cgaatctatt atgactacta aatctgtttc tttccgccgg ttgtcccatc ttaggaaact
     6001 tgtaccaggc tttgggaaag catatacaat tttcaacaaa actctcatgg aggctgacgc
     6061 gcattacaag tctgttcgga cttggaacga gatcatacca tccaaggggt gcttgcgggt
     6121 tggtggtaga tgccaccccc acgtaaacgg cgtattcttc aatggtatta tccttggccc
     6181 cgacggcaac gtgctgattc cagagatgca aagctccctt ttgcaacaac acatggagtt
     6241 gctcgtttct tccgtcatac cacttatgca tcctctcgca gaccccagta cggtattcaa
     6301 aaatggtgat gaggccgaag actttgtaga ggtccacttg ccagatgtac atgaacgaat
     6361 cagtggggta gacctcgggc ttccaaactg gggcaagtac gttcttctca gtgccggagc
     6421 gttgaccgcc ctcatgctga taatattcct catgacgtgc tggcggagag tgaacagatc
     6481 cgaaccaacc caacataacc tgcgagggac aggtcgggaa gtgtctgtaa ctccacaatc
     6541 cgggaaaatc atatcttcct gggagagcca taagtctggt ggtgagaccc gactgtaatt
     6601 tgtagcggcc gctacaaagc ttatgccgac gtacgacaat caacctctgg attacaaaat
     6661 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     6721 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     6781 gtataaatcc tggttagttc ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     6841 ctggacaggg gctcggctgt tgggcactga caattccgtg gggatccaac ctacaaacga
     6901 acctacaaac gggtggagga tcaccccacc cgacacttca caatcaaggg gtacaataca
     6961 caagggtgga ggaacacccc accctccaga cacattacac agaaatccaa tcaaacagaa
     7021 gcaccatcag ggcttctgct accaaattta tctcaaaaaa ctacaacaag gaatcaccat
     7081 cagggattcc ctgtgcaata tacgtcaaac gagggccacg acgggaggac gatcacgcct
     7141 cccgaatatc ggcatgtctg gctttcgaat tcagtgcgtg gagcatcagc ccacgcagcc
     7201 aatcagagtc gctagcaaac tggggcacaa gcgaattatc gaattcccac ctaggtaggg
     7261 atccaagggt accttcggcg cgccgtatga tcattggcta gctcgacatg tctactcgag
     7321 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     7381 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     7441 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     7501 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggctaga gcgggcccgg
     7561 tactaaccaa actggatctc tgctgtccct gtaataaacc cgaaaatttt gaatttttgt
     7621 aatttgtttt tgtaattctt tagtttgtat gtctgttgct attatgtcta ctattctttc
     7681 ccctgcactg taccccccaa tccccccttt tcttttaaaa ttgtggatga atactgccat
     7741 ttgtctgcag aattggcgca cgcagtgccg atccgttcac taatcgaatg gatctgtctc
     7801 tgtctctctc tccaccttct tcttctattc cttcgggcct gtcgggtccc ctcggggttg
     7861 ggaggtgggt ctgaaacgat aatggtgaat atccctgcct aactctattc actatagaaa
     7921 gtacagcaaa aactattctt aaacctacca agcctcctac tatcattatg aataatttta
     7981 tataccacag ccaatttgtt atgttaaacc aattccacaa acttgcccat ttatctaatt
     8041 ccaataattc ttgttcattc ttttcttgct ggttttgcga ttcttcaatt aaggagtgta
     8101 ttaagcttgt gtaattgtta atttctctgt cccactccat ccaggtcgtg tgattccaaa
     8161 tctgttccag agatttatta ctccaactag cattccaagg cacagcagtg gtgcaaatga
     8221 gttttccaga gcaaccccaa atccccagga gctgttgatc ctttaggtat ctttccacag
     8281 ccaggattct tgcctggagc tgcttgatgc cccagactgt gagttgcaac agatgctgtt
     8341 gcgcctcaat agccctcagc aaattgttct gctgctgcac tataccagac aataattgtc
     8401 tggcctgtac cgtcagcgtc attgacgctg cgcccatagt gcttcctgct gctcccaaga
     8461 acccaaggaa caaagctcct attcccactg ctcttttttc tctctgcacc actcttctct
     8521 ttgccttggt gggtgctact cctaatggtt caatttttac tactttatat ttatataatt
     8581 cacttctcca attgtccctc atatctcctc ctccaggtct gaagatcagc ggccggccgc
     8641 ttgctgtgcg gtggtcttac ttttgttttg ctcttcctct atcttgtcta aagcttcctt
     8701 ggtgtctttt atctctatcc tttgatgcac acaatagagg gttgctactg tattatataa
     8761 tgatctaagt tcttctgatc ctgtctgaag ggatggttgt agctgtccca gtatttgtct
     8821 acagccttct gatgtttcta acaggccagg attaactgcg aatcgttcta gctccctgct
     8881 tgcccatact atatgtttta atttatattt tttctttccc cctggcctta accgaatttt
     8941 ttcccatcgc gatctaattc tcccccgctt aatactgacg ctctcgcacc catctctctc
     9001 cttctagcct ccgctagtca aaatttttgg cgtactcacc agtcgccgcc cctcgcctct
     9061 tgccgtgcgc gcttcagcaa gccgagtcct gcgtcgagag agctcctctg gtttcccttt
     9121 cgctttcaag tccctgttcg ggcgccactg ctagagattt tccacactga ctaaaagggt
     9181 ctgagggatc tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact
     9241 caaggcaagc tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag
     9301 gctcagatct ggtctaacca gagagaccca gtacaggcaa aacgcgctgc ttatatagac
     9361 ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt
     9421 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
     9481 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg
     9541 catcaccatg gtaatagcga tgactaatac gtaga
//
