LOCUS       sequence-339064-        9173 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Generation of oT7pol-expressing stable cell lines, resistant to the 
            antibiotic Blasticidin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9173)
  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 9173)
  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-G_oT7pol-IRES-BSD
FEATURES             Location/Qualifiers
     source          1..9173
                     /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"
     misc_feature    5416..5878
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5472..5489)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5699..5718
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5892..6290
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             6660..7129
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(7259..7276)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7430..8018)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7510..7529)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8178..8969,8970..9038))
                     /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     8801..8820
                     /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 cccgccacca tgaacaccat caatattgcc aagaacgact tttctgatat
     2701 cgagctggcc gctattccat tcaatacact ggctgaccac tacggagagc ggctggcccg
     2761 cgaacagctg gctctggagc atgaaagcta tgagatggga gaagcccgat tcaggaagat
     2821 gtttgagagg cagctgaaag ctggggaagt ggcagacaac gcagccgcta agccactgat
     2881 taccacactg ctgcccaaaa tgatcgccag aattaatgat tggttcgagg aagtgaaggc
     2941 aaaacgagga aagaggccta ccgccttcca gtttctgcag gagatcaagc cagaagctgt
     3001 ggcatacatc accatcaaga ctaccctggc atgcctgaca agcgccgaca acacaactgt
     3061 gcaggcagtc gcttccgcaa tcggacgagc tattgaggac gaagcacgct ttgggagaat
     3121 ccgggatctg gaggccaagc acttcaagaa aaacgtggag gaacagctga acaagagggt
     3181 ggggcatgtc tataagaagg ccttcatgca ggtggtcgag gccgacatgc tgtcaaaggg
     3241 actgctggga ggagaggcct ggagctcctg gcacaaagaa gatagcatcc atgtgggagt
     3301 ccgctgcatc gagatgctga ttgaatctac tgggatggtg agtctgcacc gacagaacgc
     3361 cggcgtggtc ggacaggact ctgagacaat cgaactggct cccgagtatg ccgaagctat
     3421 tgcaactaga gccggagctc tggcagggat cagtccaatg ttccagccct gcgtggtccc
     3481 ccctaagcct tggactggca tcaccggggg cggatactgg gctaatggaa ggagaccact
     3541 ggcactggtg cgaacacact ctaagaaagc cctgatgaga tacgaggatg tctatatgcc
     3601 cgaagtgtat aaggccatca acattgctca gaatacagca tggaaaatta acaagaaagt
     3661 gctggccgtc gctaatgtga tcactaagtg gaaacattgt cccgtggagg acatccctgc
     3721 cattgaacgg gaggaactgc ctatgaagcc agaggacatc gatatgaacc ctgaagctct
     3781 gaccgcatgg aagcgcgcag ccgctgcagt gtacagaaag gataaagccc ggaagtcccg
     3841 gcgcatttct ctggagttca tgctggaaca ggccaacaag tttgctaatc acaaagcaat
     3901 ctggttcccc tacaacatgg actggcgcgg acgagtctat gccgtgagca tgttcaaccc
     3961 tcaggggaat gatatgacaa agggcctgct gactctggct aaggggaaac caattgggaa
     4021 ggagggctac tattggctga aaatccacgg ggccaattgc gctggcgtcg acaaggtgcc
     4081 attccccgag aggatcaagt tcatcgagga aaaccatgaa aatattatgg catgtgccaa
     4141 gtctcccctg gagaacacat ggtgggccga acaggatagt cctttctgct ttctggcctt
     4201 ctgttttgag tacgctggag tgcagcacca tgggctgagt tataattgct ccctgccact
     4261 ggcctttgac ggctcttgta gtggaatcca gcacttctcc gcaatgctga gggatgaggt
     4321 cggaggaaga gcagtgaacc tgctgccatc tgagacagtg caggacatct acggcattgt
     4381 cgccaagaaa gtgaatgaga tcctgcaggc tgacgcaatt aacgggactg ataatgaggt
     4441 ggtcaccgtc acagatgaaa acactggcga gatcagcgaa aaggtgaaac tgggaaccaa
     4501 ggccctggct ggacagtggc tggcatacgg agtcacccgc tcagtgacaa agcgaagcgt
     4561 gatgaccctg gcttatggca gcaaagagtt cggcttcagg cagcaggtgc tggaagacac
     4621 catccagcca gccattgatt ccggaaaggg gctgatgttt acacagccca accaggccgc
     4681 tggctacatg gccaagctga tctgggagtc agtgagcgtc acagtggtcg cagccgtgga
     4741 agctatgaat tggctgaagt ccgctgcaaa actgctggcc gctgaggtga aggacaagaa
     4801 aactggcgaa attctgagga aaagatgcgc cgtccactgg gtgacccctg atggattccc
     4861 agtgtggcag gagtataaga aacccatcca gaccagactg aacctgatgt tcctgggcca
     4921 gtttcggctg cagcctacaa tcaacactaa taaggacagt gagattgatg ctcataaaca
     4981 ggaatcaggg attgcaccta attttgtgca cagccaggac ggctcccatc tgcggaagac
     5041 tgtggtctgg gctcacgaga aatacggcat cgaatccttc gcaccgattc atgactcttt
     5101 tggaaccatc ccagccgatg cagccaacct gttcaaggct gtccgcgaga ctatggtgga
     5161 cacctacgaa agttgtgatg tgctggccga cttctatgat cagtttgctg accagctgca
     5221 cgagtcacag ctggataaga tgcccgcact gcctgccaaa ggcaacctga atctgagaga
     5281 catcctggag tccgatttcg catttgcctg actcgacccc ccccctaacg ttactggccg
     5341 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     5401 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     5461 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     5521 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     5581 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     5641 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     5701 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     5761 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     5821 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataagc
     5881 ttgccacaac catggccaag cctttgtctc aagaagaatc caccctcatt gaaagagcaa
     5941 cggctacaat caacagcatc cccatctctg aagactacag cgtcgccagc gcagctctct
     6001 ctagcgacgg ccgcatcttc actggtgtca atgtatatca ttttactggg ggaccttgtg
     6061 cagaactcgt ggtgctgggc actgctgctg ctgcggcagc tggcaacctg acttgtatcg
     6121 tcgcgatcgg aaatgagaac aggggcatct tgagcccctg cggacggtgc cgacaggtgc
     6181 ttctcgatct gcatcctggg atcaaagcca tagtgaagga cagtgatgga cagccgacgg
     6241 cagttgggat tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa gcacttcgtg
     6301 gccgaggagc aggactgaca cgacccggac gggacgctcc tgcgcctgat acagaacgaa
     6361 ttgcttgcag gcatctcatg agtgtgtctt cccgttttcc gcctgaggtc actgcgtgga
     6421 tggagcatca gtggtccagg ctctagtttt gactcaacaa tatcaccagc tgaagcctat
     6481 agagtacgag ccatagataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     6541 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     6601 gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc
     6661 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     6721 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     6781 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
     6841 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     6901 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     6961 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt
     7021 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     7081 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac atgcagcatg
     7141 tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt
     7201 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     7261 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     7321 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     7381 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     7441 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     7501 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     7561 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     7621 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7681 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7741 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7801 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7861 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7921 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7981 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     8041 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     8101 caaaaaggat cttcacctag atccttttgc ggccgcaaat caatctaaag tatatatgag
     8161 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8221 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8281 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8341 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8401 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8461 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8521 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8581 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8641 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8701 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8761 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8821 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8881 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8941 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9001 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatgatcc
     9061 ggccattagc catattattc attggttata tagcataaat caatattggc tattggccat
     9121 tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt cca
//
