LOCUS       sequence-338971-        8477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Generation of stable cell lines expressing the rabies optimized 
            glycoprotein (oG), resistant to the antibiotic Puromycin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8477)
  AUTHORS   Sumser A, Joesch M, Jonas P, Ben-Simon Y
  TITLE     Fast, high-throughput production of improved rabies viral vectors 
            for specific, efficient and versatile transsynaptic retrograde 
            labeling.
  JOURNAL   Elife. 2022 Aug 30;11:e79848. doi: 10.7554/eLife.79848.
  PUBMED    36040301
REFERENCE   2  (bases 1 to 8477)
  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: "Elife."; 
            date: "2022-08-30"; volume: "11:e79848. doi"; pages: " 
            10.7554/eLife.79848"
COMMENT     Sequence Label: MMLV-CAG-SADB19_oG-IRES-Puro
FEATURES             Location/Qualifiers
     source          1..8477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        17..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        397..600
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     551..571
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             601..776
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    839..1038
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1239..1655
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1585..1607
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1623..1639
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     2517..2539
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2601..2620
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2659..2668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    4374..4836
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4430..4447)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4657..4676
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4869..5468
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4869..4888)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5365..5385
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5964..6433
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6563..6580)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6734..7322)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6814..6833)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7482..8273,8274..8342))
                     /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     8105..8124
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 acattaccgc catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
       61 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      121 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      181 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      241 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      301 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      361 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      421 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      481 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      541 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      601 caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc
      661 cgggtacccg tattcccaat aaagcctctt gctgtttgca tccgaatcgt ggtctcgctg
      721 ttccttggga gggtctcctc tgagtgattg actacccacg acgggggtct ttcatttggg
      781 ggctcgtccg ggatttggag acccctgccc agggaccacc gacccaccac cgggaggtaa
      841 gctggccagc aacttatctg tgtctgtccg attgtctagt gtctatgttt gatgttatgc
      901 gcctgcgtct gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg
      961 acgagttctg aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt
     1021 tttgtggccc gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg
     1081 ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg
     1141 tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc
     1201 tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct
     1261 taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag
     1321 atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg
     1381 gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct
     1441 tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct
     1501 tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc
     1561 ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct
     1621 ccctttatcc agccctcact ccttctctag gcgccggaat tgcagagagg gagtggccaa
     1681 ctccatcact aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta
     1741 gccatgctct aggaagagta ccattgacgt caataatgac gtatgttccc atagtaacgc
     1801 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     1861 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     1921 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc
     2041 tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt
     2101 gtgcagcgat gggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc
     2161 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
     2221 cgaagcgcgc gacgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc
     2281 gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg
     2341 cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct
     2401 tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc
     2461 ggctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt
     2521 cggcttctgg cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt
     2581 ctttttccta cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa
     2641 agaattggat cctctagagc caccatggtt cctcaggttc ttttgtttgt actccttctg
     2701 gggttttcgt tgtgtttcgg gaagttcccc atttacacga taccagacaa acttggtccc
     2761 tggagcccta ttgacataca ccatctccgc tgtccaaata acctggttgt ggaggatgaa
     2821 ggatgtacca acctgtccgg gttctcctat atggaactca aagtgggata catctcagcc
     2881 atcaaagtga acgggttcac ttgcacaggt gttgtgacag aggcagagac ctacaccaac
     2941 tttgttggtt atgtcacaac cacattcaag agaaagcatt tccgccccac cccagacgca
     3001 tgtagagccg cgtataactg gaagatggcc ggtgacccca gatatgaaga gtccctacaa
     3061 aatccatacc ccgactacca ctggcttcga actgtaagaa ccaccaaaga gtccctcatt
     3121 atcatatccc caagtgtgac agatttggac ccatatgaca aatcccttca ctcaagggtc
     3181 ttccctggcg gaaagtgctc aggaataacg gtgtcctcta cctactgctc aactaaccat
     3241 gattacacca tttggatgcc cgaggacccg agaccaggga caccttgtga catttttacc
     3301 aatagcagag ggaagagagc atccaacggg aacaagactt gcggctttgt ggatgaaaga
     3361 ggcctgtata agtctctaaa aggagcatgc aggctcaagt tatgtggagt tcttggactt
     3421 agacttatgg atggaacatg ggtcgcgatg caaacatcag atgagaccaa atggtgctct
     3481 ccagatcagt tggtgaattt gcacgacttt cgctcaaacg agattgagca tctcgttgtg
     3541 gaggatttag tcaagaaaag agaggaatgt ctggatacat tagagtccat catgaccacc
     3601 aagtcagtaa gtttcagacg tctcagtcac ctgagaaaac ttgtcccagg gtttggaaaa
     3661 gcatatacca tattcaacaa aaccttgatg gaggctgatg ttcactacaa gtcagtccgg
     3721 acctggaatg agatcatccc ctcaaaaggg tgtttgaaag ttggaggaag gtgccatcct
     3781 catgtgaacg gggtgttttt caatggtata atattagggc ctgacgaccg tgtcctaatc
     3841 ccagagatgc aatcatccct cctccggcaa catatggagt tgttggaatc ttcagttatc
     3901 cccctgatgc accccctggc tgacccttct acagttttca aagaaggtga tgaggctgag
     3961 gattttgttg aagttcacct ccccgatgtg tacaaaaaga tctcaggggt tgacctgggt
     4021 ctcccgaact ggggaaagta tgtattgatg actgcagggg ccatgattgg cctggtgttg
     4081 atattttccc taatgacatg gtgcagaaga gccaatcgac cagaatcgaa acaacgcagt
     4141 tttggaggga caggggggaa tgtgtcagtc acttcccaaa gcggaaaagt cataccttca
     4201 tgggaatcat ataagagtgg aggtgagatc agactgtaag aattccsmcc cccccccccc
     4261 cccccnycyc ccyccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg
     4321 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc
     4381 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa
     4441 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga
     4501 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc
     4561 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc
     4621 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac
     4681 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg
     4741 tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac
     4801 ggggacgtgg ttttcctttg aaaaacacga tgataagctt gccacaaccc acaaggagac
     4861 gaccttccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc
     4921 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
     4981 acccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
     5041 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
     5101 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
     5161 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
     5221 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
     5281 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
     5341 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
     5401 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
     5461 gtgcctgacg cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg
     5521 gctccgaccg aagccacccg gggcggcccc gccgaccccg cacccgcccc cgaggcccac
     5581 cgactctagt cgagggctgc agcgctgcag aggccgagtg cagaactgct ccaaagggac
     5641 ctcaaggctt tccgagggac actaggctga ctccatcgag ccagtgtaga gataagctta
     5701 tcgattagtc caatttgtta aagacaggat atcagtggtc caggctctag ttttgactca
     5761 acaatatcac cagctgaagc ctatagagta cgagccatag ataaaataaa agattttatt
     5821 tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta
     5881 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga agttcagatc
     5941 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
     6001 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
     6061 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
     6121 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     6181 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     6241 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc
     6301 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     6361 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     6421 cgggggtctt tcacatgcag catgtatcaa aattaatttg gttttttttc ttaagtattt
     6481 acattaaatg gccatagttg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6541 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6601 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6661 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6721 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6781 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6841 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6901 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6961 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7021 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7081 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7141 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7201 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7261 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7321 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7381 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttgcggccgc
     7441 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7501 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7561 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7621 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7681 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7741 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7801 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7861 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7921 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7981 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     8041 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     8101 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     8161 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     8221 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     8281 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     8341 atactcttcc tttttcaatg atccggccat tagccatatt attcattggt tatatagcat
     8401 aaatcaatat tggctattgg ccattgcata cgttgtatcc atatcataat atgtacattt
     8461 atattggctc atgtcca
//
