LOCUS       sequence-419147-        9473 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Packaging plasmid for v3b PE-eVLP production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9473)
  AUTHORS   An M, Raguram A, Du SW, Banskota S, Davis JR, Newby GA, Chen PZ, 
            Palczewski K, Liu DR
  TITLE     Engineered virus-like particles for transient delivery of prime 
            editor ribonucleoprotein complexes in vivo.
  JOURNAL   Nat Biotechnol. 2024 Jan 8. doi: 10.1038/s41587-023-02078-y.
  PUBMED    38191664
REFERENCE   2  (bases 1 to 9473)
  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: "Nat 
            Biotechnol. 2024 Jan 8. doi: 10.1038/s41587-023-02078-y."
COMMENT     Sequence Label: gag-P3-pol
FEATURES             Location/Qualifiers
     source          1..9473
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        96..475
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        476..679
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     630..650
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     676..700
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             837..1253
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1183..1205
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1221..1237
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    5760..6134
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     6072..6088
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     primer_bind     complement(6269..6286)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6275..6499
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6573..6592)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6574..6592)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6599..6616)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6599..6615)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6608..6630)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6757..7212
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6844..6863)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7054..7075
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7238..7342
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7343..7411,7412..8203)
                     /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(7561..7580)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8374..8962
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8863..8882
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9116..9133
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9250..9271
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9286..9291,9292..9309,9310..9316)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9324..9340
                     /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)."
     primer_bind     9329..9351
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9348..9364
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9348..9364
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        9385..9403
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ataaatcaat attggctatt ggccattgca tacgttgtat ccatatcata atatgtacat
       61 ttatattggc tcatgtccaa cattaccgcc atgttgacat tgattattga ctagttatta
      121 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      181 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      241 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      301 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      361 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      421 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      481 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      541 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      601 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      661 ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac gccatccacg
      721 ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccgcggcc ccaagcttgg
      781 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aatatgggcc
      841 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
      901 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
      961 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
     1021 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
     1081 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
     1141 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
     1201 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gccaaaccta
     1261 aacctcaagt tctttctgac agtggggggc cgctcatcga cctacttaca gaagaccccc
     1321 cgccttatag ggacccaaga ccaccccctt ccgacaggga cggaaatggt ggagaagcga
     1381 cccctgcggg agaggcaccg gacccctccc caatggcatc tcgcctacgt gggagacggg
     1441 agccccctgt ggccgactcc actacctcgc aggcattccc cctccgcgca ggaggaaacg
     1501 gacagcttca atactggccg ttctcctctt ctgaccttta caactggaaa aataataacc
     1561 cttctttttc tgaagatcca ggtaaactga cagctctgat cgagtctgtc ctcatcaccc
     1621 atcagcccac ctgggacgac tgtcagcagc tgttggggac tctgctgacc ggagaagaaa
     1681 aacaacgggt gctcttagag gctagaaagg cggtgcgggg cgatgatggg cgccccactc
     1741 aactgcccaa tgaagtcgat gccgcttttc ccctcgagcg cccagactgg gattacacca
     1801 cccaggcagg taggaaccac ctagtccact atcgccagtt gctcctagcg ggtctccaaa
     1861 acgcgggcag aagccccacc aatttggcca aggtaaaagg aataacacaa gggcccaatg
     1921 agtctccctc ggccttccta gagagactta aggaagccta tcgcaggtac actccttatg
     1981 accctgagga cccagggcaa gaaactaatg tgtctatgtc tttcatttgg cagtctgccc
     2041 cagacattgg gagaaagtta gagaggttag aagatttaaa aaacaagacg cttggagatt
     2101 tggttagaga ggcagaaaag atctttaata aacgagaaac cccggaagaa agagaggaac
     2161 gtatcaggag agaaacagag gaaaaagaag aacgccgtag gacagaggat gagcagaaag
     2221 agaaagaaag agatcgtagg agacatagag agatgagcaa gctattggcc actgtcgtta
     2281 gtggacagaa acaggataga cagggaggag aacgaaggag gtcccaactc gatcgcgacc
     2341 agtgtgccta ctgcaaagaa aaggggcact gggctaaaga ttgtcccaag aaaccacgag
     2401 gacctcgggg accaagaccc cagacctccc tcctgaccct agatgactcc ggtggaggtg
     2461 gatccggtgg aggttcctcc ccggaagacg aaatccaaca gttggaggaa gagatcgcac
     2521 aacttgagca gaaaaacgca gcactcaaag agaaaaacca agcactgaag tacgggtccg
     2581 gaggtggagg tggataggga ggtcagggtc aggagccccc ccctgaaccc aggataaccc
     2641 tcaaagtcgg ggggcaaccc gtcaccttcc tggtagatac tggggcccaa cactccgtgc
     2701 tgacccaaaa tcctggaccc ctaagtgata agtctgcctg ggtccaaggg gctactggag
     2761 gaaagcggta tcgctggacc acggatcgca aagtacatct agctaccggt aaggtcaccc
     2821 actctttcct ccatgtacca gactgtccct atcctctgtt aggaagagat ttgctgacta
     2881 aactaaaagc ccaaatccac tttgagggat caggagctca ggttatggga ccaatggggc
     2941 agcccctgca agtgttgacc ctaaatatag aagatgagta tcggctacat gagacctcaa
     3001 aagagccaga tgtttctcta gggtccacat ggctgtctga ttttcctcag gcctgggcgg
     3061 aaaccggggg catgggactg gcagttcgcc aagctcctct gatcatacct ctgaaagcaa
     3121 cctctacccc cgtgtccata aaacaatacc ccatgtcaca agaagccaga ctggggatca
     3181 agccccacat acagagactg ttggaccagg gaatactggt accctgccag tccccctgga
     3241 acacgcccct gctacccgtt aagaaaccag ggactaatga ttataggcct gtccaggatc
     3301 tgagagaagt caacaagcgg gtggaagaca tccaccccac cgtgcccaac ccttacaacc
     3361 tcttgagcgg gctcccaccg tcccaccagt ggtacactgt gcttgattta aaggatgcct
     3421 ttttctgcct gagactccac cccaccagtc agcctctctt cgcctttgag tggagagatc
     3481 cagagatggg aatctcagga caattgacct ggaccagact cccacagggt ttcaaaaaca
     3541 gtcccaccct gtttgatgag gcactgcaca gagacctagc agacttccgg atccagcacc
     3601 cagacttgat cctgctacag tacgtggatg acttactgct ggccgccact tctgagctag
     3661 actgccaaca aggtactcgg gccctgttac aaaccctagg gaacctcggg tatcgggcct
     3721 cggccaagaa agcccaaatt tgccagaaac aggtcaagta tctggggtat cttctaaaag
     3781 agggtcagag atggctgact gaggccagaa aagagactgt gatggggcag cctactccga
     3841 agacccctcg acaactaagg gagttcctag ggacggcagg cttctgtcgc ctctggatcc
     3901 ctgggtttgc agaaatggca gcccccttgt accctctcac caaaacgggg actctgttta
     3961 attggggccc agaccaacaa aaggcctatc aagaaatcaa gcaagctctt ctaactgccc
     4021 cagccctggg gttgccagat ttgactaagc cctttgaact ctttgtcgac gagaagcagg
     4081 gctacgccaa aggtgtccta acgcaaaaac tgggaccttg gcgtcggccg gtggcctacc
     4141 tgtccaaaaa gctagaccca gtagcagctg ggtggccccc ttgcctacgg atggtagcag
     4201 ccattgccgt actgacaaag gatgcaggca agctaaccat gggacagcca ctagtcattc
     4261 tggcccccca tgcagtagag gcactagtca aacaaccccc cgaccgctgg ctttccaacg
     4321 cccggatgac tcactatcag gccttgcttt tggacacgga ccgggtccag ttcggaccgg
     4381 tggtagccct gaacccggct acgctgctcc cactgcctga ggaagggctg caacacaact
     4441 gccttgatat cctggccgaa gcccacggaa cccgacccga cctaacggac cagccgctcc
     4501 cagacgccga ccacacctgg tacacggatg gaagcagtct cttacaagag ggacagcgta
     4561 aggcgggagc tgcggtgacc accgagaccg aggtaatctg ggctaaagcc ctgccagccg
     4621 ggacatccgc tcagcgggct gaactgatag cactcaccca ggccctaaag atggcagaag
     4681 gtaagaagct aaatgtttat actgatagcc gttatgcttt tgctactgcc catatccatg
     4741 gagaaatata cagaaggcgt gggttgctca catcagaagg caaagagatc aaaaataaag
     4801 acgagatctt ggccctacta aaagccctct ttctgcccaa aagacttagc ataatccatt
     4861 gtccaggaca tcaaaaggga cacagcgccg aggctagagg caaccggatg gctgaccaag
     4921 cggcccgaaa ggcagccatc acagagactc cagacacctc taccctcctc atagaaaatt
     4981 catcacccta cacctcagaa cattttcatt acacagtgac tgatataaag gacctaacca
     5041 agttgggggc catttatgat aaaacaaaga agtattgggt ctaccaagga aaacctgtga
     5101 tgcctgacca gtttactttt gaattattag actttcttca tcagctgact cacctcagct
     5161 tctcaaaaat gaaggctctc ctagagagaa gccacagtcc ctactacatg ctgaaccggg
     5221 atcgaacact caaaaatatc actgagacct gcaaagcttg tgcacaagtc aacgccagca
     5281 agtctgccgt taaacaggga actagggtcc gcgggcatcg gcccggcact cattgggaga
     5341 tcgatttcac cgagataaag cccggattgt atggctataa atatcttcta gtttttatag
     5401 ataccttttc tggctggata gaagccttcc caaccaagaa agaaaccgcc aaggtcgtaa
     5461 ccaagaagct actagaggag atcttcccca ggttcggcat gcctcaggta ttgggaactg
     5521 acaatgggcc tgccttcgtc tccaaggtga gtcagacagt ggccgatctg ttggggattg
     5581 attggaaatt acattgtgca tacagacccc aaagctcagg ccaggtagaa agaatgaata
     5641 gaaccatcaa ggagacttta actaaattaa cgcttgcaac tggctctaga gactgggtgc
     5701 tcctactccc cttagccctg taccgagccc gcaacacgcc gggcccccat ggcctcaccc
     5761 catatgagat cttatatggg gcacccccgc cccttgtaaa cttccctgac cctgacatga
     5821 caagagttac taacagcccc tctctccaag ctcacttaca ggctctctac ttagtccagc
     5881 acgaagtctg gagacctctg gcggcagcct accaagaaca actggaccga ccggtggtac
     5941 ctcaccctta ccgagtcggc gacacagtgt gggtccgccg acaccagact aagaacctag
     6001 aacctcgctg gaaaggacct tacacagtcc tgctgaccac ccccaccgcc ctcaaagtag
     6061 acggcatcgc agcttggata cacgccgccc acgtgaaggc tgccgacccc gggggtggac
     6121 catcctctag actgacatgg cgcgttcaac gctctcaaaa ccccttaaaa ataaggttga
     6181 cgcgtcaacc cgcgaggccc cctaatcccc ttaattcttc tgatgctcag aggggtcagt
     6241 atcgaattcc tgcagccccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     6301 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     6361 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     6421 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     6481 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc gatcctctag
     6541 agcggccgcc accgcggtgg agctccaatt cgccctatag tgagtcgtat tacaattcac
     6601 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     6661 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     6721 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     6781 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     6841 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     6901 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     6961 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     7021 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     7081 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     7141 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     7201 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     7261 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     7321 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     7381 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     7441 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     7501 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     7561 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     7621 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     7681 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7741 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     7801 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7861 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     7921 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     7981 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     8041 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     8101 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     8161 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     8221 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     8281 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     8341 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     8401 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     8461 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     8521 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     8581 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     8641 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     8701 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     8761 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     8821 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     8881 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     8941 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     9001 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     9061 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     9121 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     9181 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     9241 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     9301 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     9361 tgaccatgat tacgccaagc tcgaaattaa ccctcactaa agggaacaaa agctggtacc
     9421 gggccccccc tcgagatccg gccattagcc atattattca ttggttatat agc
//
