LOCUS       sequence-96845-e        8895 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gag/pol packaging plasmid for production of lentiviral vectors with 
            mutated reverse transcriptase (NRTLV, non-reverse transcribable 
            lentiviral vectors) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8895)
  AUTHORS   Mock U, Riecken K, Berdien B, Qasim W, Chan E, Cathomen T, Fehse B
  TITLE     Novel lentiviral vectors with mutated reverse transcriptase for mRNA
            delivery of TALE nucleases.
  JOURNAL   Sci Rep. 2014 Sep 18;4:6409. doi: 10.1038/srep06409.
  PUBMED    25230987
REFERENCE   2  (bases 1 to 8895)
  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: "Sci Rep."; 
            date: "2014-09-18"; volume: "4:6409. doi"; pages: " 
            10.1038/srep06409"
COMMENT     Sequence Label: dRT-pMDLg/pRRE
FEATURES             Location/Qualifiers
     source          1..8895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(40..59)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        161..540
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        541..744
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     695..715
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     741..765
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          878..1353
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     regulatory      1431..1440
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1437..2945
                     /codon_start=1
                     /gene="gag"
                     /product="gag protein from human immunodeficiency virus 1"
                     /label=HIV-1 gag
                     /translation="MGARASVLSGGELDRWEKIRLRPGGKKKYKLKHIVWASRELERFA
                     VNPGLLETSEGCRQILGQLQPSLQTGSEELRSLYNTVATLYCVHQRIEIKDTKEALDKI
                     EEEQNKSKKKAQQAAADTGHSNQVSQNYPIVQNIQGQMVHQAISPRTLNAWVKVVEEKA
                     FSPEVIPMFSALSEGATPQDLNTMLNTVGGHQAAMQMLKETINEEAAEWDRVHPVHAGP
                     IAPGQMREPRGSDIAGTTSTSTLQEQIGWMTHNPPIPVGEIYKRWIILGLNKIVRMYSP
                     TSILDIRQGPKEPFRDYVDRFYKTLRAEQASQEVKNWMTETLLVQNANPDCKTILKALG
                     PGATLEEMMTACQGVGGPGHKARVLAEAMSQVTNPATIMIQKGNFRNQRKTVKCFNCGK
                     EGHIAKNCRAPRKKGCWKCGKEGHQMKDCTERQANFLGKIWPSHKGRPGNFLQSRPEPT
                     APPEESFRFGEETTTPSQKQEPIDKELYPLASLRSLFGSDPSSQ"
     CDS             2738..5749
                     /codon_start=1
                     /gene="pol"
                     /product="pol protein from human immunodeficiency virus 1"
                     /label=HIV-1 pol
                     /translation="FFREDLAFPQGKAREFSSEQTRANSPTRRELQVWGRDNNSLSEAG
                     ADRQGTVSFSFPQITLWQRPLVTIKIGGQLKEALLDTGADDTVLEEMNLPGRWKPKMIG
                     GIGGFIKVGQYDQILIEICGHKAIGTVLVGPTPVNIIGRNLLTQIGCTLNFPISPIETV
                     PVKLKPGMDGPKVKQWPLTEEKIKALVEICTEMEKEGKISKIGPENPYNTPVFAIKKKD
                     STKWRKLVDFRELNKRTQDFWEVQLGIPHPAGLKQKKSVTVLDVGDAYFSVPLDKDFRK
                     YTAFTIPSINNETPGIRYQYNVLPQGWKGSPAIFQCSMTKILEPFRKQNPDIVIYQYMV
                     VLYVGSDLEIGQHRTKIEELRQHLLRWGFTTPDKKHQKEPPFLWMGYELHPDKWTVQPI
                     VLPEKDSWTVNDIQKLVGKLNWASQIYAGIKVRQLCKLLRGTKALTEVVPLTEEAELEL
                     AENREILKEPVHGVYYDPSKDLIAEIQKQGQGQWTYQIYQEPFKNLKTGKYARMKGAHT
                     NDVKQLTEAVQKIATESIVIWGKTPKFKLPIQKETWEAWWTEYWQATWIPEWEFVNTPP
                     LVKLWYQLEKEPIIGAETFYVDGAANRETKLGKAGYVTDRGRQKVVPLTDTTNQKTELQ
                     AIHLALQDSGLEVNIVTDSQYALGIIQAQPDKSESELVSQIIEQLIKKEKVYLAWVPAH
                     KGIGGNEQVDGLVSAGIRKVLFLDGIDKAQEEHEKYHSNWRAMASDFNLPPVVAKEIVA
                     SCDKCQLKGEAMHGQVDCSPGIWQLDCTHLEGKVILVAVHVASGYIEAEVIPAETGQET
                     AYFLLKLAGRWPVKTVHTDNGSNFTSTTVKAACWWAGIKQEFGIPYNPQSQGVIESMNK
                     ELKKIIGQVRDQAEHLKTAVQMAVFIHNFKRKGGIGGYSAGERIVDIIATDIQTKELQK
                     QITKIQNFRVYYRDSRDPVWKGPAKLLWKGEGAVVIQDNSDIKVVPRRKAKIIRDYGKQ
                     MAGDDCVASRQDED"
     misc_feature    5434..5551
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5774..6007
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     polyA_signal    6238..6631
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     primer_bind     complement(6970..6987)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7141..7729)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7221..7240)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7900..8691,8692..8760))
                     /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     8523..8542
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8761..8865)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggatcccctg agggggcccc catgggctag aggatccggc ctcggcctct gcataaataa
       61 aaaaaattag tcagccatga gcttggccca ttgcatacgt tgtatccata tcataatatg
      121 tacatttata ttggctcatg tccaacatta ccgccatgtt gacattgatt attgactagt
      181 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      241 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      301 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      361 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      421 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      481 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      541 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      601 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      661 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      721 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      781 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccc ctcgaagctt
      841 acatgtggta ccgagctcgg atcctgagaa cttcagggtg agtctatggg acccttgatg
      901 ttttctttcc ccttcttttc tatggttaag ttcatgtcat aggaagggga gaagtaacag
      961 ggtacacata ttgaccaaat cagggtaatt ttgcatttgt aattttaaaa aatgctttct
     1021 tcttttaata tacttttttg tttatcttat ttctaatact ttccctaatc tctttctttc
     1081 agggcaataa tgatacaatg tatcatgcct ctttgcacca ttctaaagaa taacagtgat
     1141 aatttctggg ttaaggcaat agcaatattt ctgcatataa atatttctgc atataaattg
     1201 taactgatgt aagaggtttc atattgctaa tagcagctac aatccagcta ccattctgct
     1261 tttattttat ggttgggata aggctggatt attctgagtc caagctaggc ccttttgcta
     1321 atcatgttca tacctcttat cttcctccca cagctcctgg gcaacgtgct ggtctgtgtg
     1381 ctggcccatc actttggcaa agcacgtgag atctgaattc gagatctgcc gccgccatgg
     1441 gtgcgagagc gtcagtatta agcgggggag aattagatcg atgggaaaaa attcggttaa
     1501 ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag
     1561 aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg
     1621 gacagctaca accatccctt cagacaggat cagaagaact tagatcatta tataatacag
     1681 tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag
     1741 acaagataga ggaagagcaa aacaaaagta agaaaaaagc acagcaagca gcagctgaca
     1801 caggacacag caatcaggtc agccaaaatt accctatagt gcagaacatc caggggcaaa
     1861 tggtacatca ggccatatca cctagaactt taaatgcatg ggtaaaagta gtagaagaga
     1921 aggctttcag cccagaagtg atacccatgt tttcagcatt atcagaagga gccaccccac
     1981 aagatttaaa caccatgcta aacacagtgg ggggacatca agcagccatg caaatgttaa
     2041 aagagaccat caatgaggaa gctgcagaat gggatagagt gcatccagtg catgcagggc
     2101 ctattgcacc aggccagatg agagaaccaa ggggaagtga catagcagga actactagta
     2161 ctagtaccct tcaggaacaa ataggatgga tgacacataa tccacctatc ccagtaggag
     2221 aaatctataa aagatggata atcctgggat taaataaaat agtaagaatg tatagcccta
     2281 ccagcattct ggacataaga caaggaccaa aggaaccctt tagagactat gtagaccgat
     2341 tctataaaac tctaagagcc gagcaagctt cacaagaggt aaaaaattgg atgacagaaa
     2401 ccttgttggt ccaaaatgcg aacccagatt gtaagactat tttaaaagca ttgggaccag
     2461 gagcgacact agaagaaatg atgacagcat gtcagggagt ggggggaccc ggccataaag
     2521 caagagtttt ggctgaagca atgagccaag taacaaatcc agctaccata atgatacaga
     2581 aaggcaattt taggaaccaa agaaagactg ttaagtgttt caattgtggc aaagaagggc
     2641 acatagccaa aaattgcagg gcccctagga aaaagggctg ttggaaatgt ggaaaggaag
     2701 gacaccaaat gaaagattgt actgagagac aggctaattt tttagggaag atctggcctt
     2761 cccacaaggg aaggccaggg aattttcttc agagcagacc agagccaaca gccccaccag
     2821 aagagagctt caggtttggg gaagagacaa caactccctc tcagaagcag gagccgatag
     2881 acaaggaact gtatccttta gcttccctca gatcactctt tggcagcgac ccctcgtcac
     2941 aataaagata ggggggcaat taaaggaagc tctattagat acaggagcag atgatacagt
     3001 attagaagaa atgaatttgc caggaagatg gaaaccaaaa atgatagggg gaattggagg
     3061 ttttatcaaa gtaggacagt atgatcagat actcatagaa atctgcggac ataaagctat
     3121 aggtacagta ttagtaggac ctacacctgt caacataatt ggaagaaatc tgttgactca
     3181 gattggctgc actttaaatt ttcccattag tcctattgag actgtaccag taaaattaaa
     3241 gccaggaatg gatggcccaa aagttaaaca atggccattg acagaagaaa aaataaaagc
     3301 attagtagaa atttgtacag aaatggaaaa ggaaggaaaa atttcaaaaa ttgggcctga
     3361 aaatccatac aatactccag tatttgccat aaagaaaaaa gacagtacta aatggagaaa
     3421 attagtagat ttcagagaac ttaataagag aactcaagat ttctgggaag ttcaattagg
     3481 aataccacat cctgcagggt taaaacagaa aaaatcagta acagtactgg atgtgggcga
     3541 tgcatatttt tcagttccct tagataaaga cttcaggaag tatactgcat ttaccatacc
     3601 tagtataaac aatgagacac cagggattag atatcagtac aatgtgcttc cacagggatg
     3661 gaaaggatca ccagcaatat tccagtgtag catgacaaaa atcttagagc cttttagaaa
     3721 acaaaatcca gacatagtca tctatcaata catggtggtg ttgtatgtag gatctgactt
     3781 agaaataggg cagcatagaa caaaaataga ggaactgaga caacatctgt tgaggtgggg
     3841 atttaccaca ccagacaaaa aacatcagaa agaacctcca ttcctttgga tgggttatga
     3901 actccatcct gataaatgga cagtacagcc tatagtgctg ccagaaaagg acagctggac
     3961 tgtcaatgac atacagaaat tagtgggaaa attgaattgg gcaagtcaga tttatgcagg
     4021 gattaaagta aggcaattat gtaaacttct taggggaacc aaagcactaa cagaagtagt
     4081 accactaaca gaagaagcag agctagaact ggcagaaaac agggagattc taaaagaacc
     4141 ggtacatgga gtgtattatg acccatcaaa agacttaata gcagaaatac agaagcaggg
     4201 gcaaggccaa tggacatatc aaatttatca agagccattt aaaaatctga aaacaggaaa
     4261 atatgcaaga atgaagggtg cccacactaa tgatgtgaaa caattaacag aggcagtaca
     4321 aaaaatagcc acagaaagca tagtaatatg gggaaagact cctaaattta aattacccat
     4381 acaaaaggaa acatgggaag catggtggac agagtattgg caagccacct ggattcctga
     4441 gtgggagttt gtcaataccc ctcccttagt gaagttatgg taccagttag agaaagaacc
     4501 cataatagga gcagaaactt tctatgtaga tggggcagcc aatagggaaa ctaaattagg
     4561 aaaagcagga tatgtaactg acagaggaag acaaaaagtt gtccccctaa cggacacaac
     4621 aaatcagaag actgagttac aagcaattca tctagctttg caggattcgg gattagaagt
     4681 aaacatagtg acagactcac aatatgcatt gggaatcatt caagcacaac cagataagag
     4741 tgaatcagag ttagtcagtc aaataataga gcagttaata aaaaaggaaa aagtctacct
     4801 ggcatgggta ccagcacaca aaggaattgg aggaaatgaa caagtagatg ggttggtcag
     4861 tgctggaatc aggaaagtac tatttttaga tggaatagat aaggcccaag aagaacatga
     4921 gaaatatcac agtaattgga gagcaatggc tagtgatttt aacctaccac ctgtagtagc
     4981 aaaagaaata gtagccagct gtgataaatg tcagctaaaa ggggaagcca tgcatggaca
     5041 agtagactgt agcccaggaa tatggcagct agattgtaca catttagaag gaaaagttat
     5101 cttggtagca gttcatgtag ccagtggata tatagaagca gaagtaattc cagcagagac
     5161 agggcaagaa acagcatact tcctcttaaa attagcagga agatggccag taaaaacagt
     5221 acatacagac aatggcagca atttcaccag tactacagtt aaggccgcct gttggtgggc
     5281 ggggatcaag caggaatttg gcattcccta caatccccaa agtcaaggag taatagaatc
     5341 tatgaataaa gaattaaaga aaattatagg acaggtaaga gatcaggctg aacatcttaa
     5401 gacagcagta caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg
     5461 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
     5521 acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc
     5581 agtttggaaa ggaccagcaa agctcctctg gaaaggtgaa ggggcagtag taatacaaga
     5641 taatagtgac ataaaagtag tgccaagaag aaaagcaaag atcatcaggg attatggaaa
     5701 acagatggca ggtgatgatt gtgtggcaag tagacaggat gaggattaac acatggaatt
     5761 ccggagcggc cgcaggagct ttgttccttg ggttcttggg agcagcagga agcactatgg
     5821 gcgcagcctc aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc
     5881 agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct
     5941 ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac
     6001 agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga
     6061 atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt
     6121 gggacagaga aattaacaat tacacaagct tccgcggaat tcaccccacc agtgcaggct
     6181 gcctatcaga aagtggtggc tggtgtggct aatgccctgg cccacaagtt tcactaagct
     6241 cgcttccttg ctgtccaatt tctattaaag gttccttggt tccctaagtc caactactaa
     6301 actgggggat attatgaagg gccttgagca tctggattct gcctaataaa aaacatttat
     6361 tttcattgca atgatgtatt taaattattt ctgaatattt tactaaaaag ggaatgtggg
     6421 aggtcagtgc atttaaaaca taaagaaatg aagagctagt tcaaaccttg ggaaaataca
     6481 ctatatctta aactccatga aagaaggtga ggctgcaaac agctaatgca cattggcaac
     6541 agccctgatg cctatgcctt attcatccct cagaaaagga ttcaagtaga ggcttgattt
     6601 ggaggttaaa gtttggctat gctgtatttt acattactta ttgttttagc tgtcctcatg
     6661 aatgtctttt cactacccat ttgcttatcc tgcatctctc agccttgact ccactcagtt
     6721 ctcttgctta gagataccac ctttcccctg aagtgttcct tccatgtttt acggcgagat
     6781 ggtttctcct cgcctggcca ctcagcctta gttgtctctg ttgtcttata gaggtctact
     6841 tgaagaagga aaaacagggg gcatggtttg actgtcctgt gagcccttct tccctgcctc
     6901 ccccactcac agtgacccgg aatccctcga catggcagtc tagcactagt gcggccgcag
     6961 atctgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     7021 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     7081 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7141 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7201 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7261 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7321 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7381 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7441 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7501 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7561 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     7621 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7681 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7741 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7801 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7861 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7921 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7981 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     8041 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     8101 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8161 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     8221 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8281 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8341 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8401 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8461 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8521 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8581 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8641 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8701 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8761 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8821 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8881 agtgccacct gacgt
//
