LOCUS       sequence-451028-        9326 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MMLVgag-pro-pol(Q226P) for producing v5 eVLPs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9326)
  AUTHORS   Raguram A, An M, Chen PZ, Liu DR
  TITLE     Directed evolution of engineered virus-like particles with improved 
            production and transduction efficiencies.
  JOURNAL   Nat Biotechnol. 2024 Nov 13. doi: 10.1038/s41587-024-02467-x.
  PUBMED    39537813
REFERENCE   2  (bases 1 to 9326)
  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 Nov 13. doi: 10.1038/s41587-024-02467-x."
COMMENT     Sequence Label: v5 eVLP gag-pro-pol
FEATURES             Location/Qualifiers
     source          1..9326
                     /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    5613..5987
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     5925..5941
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     primer_bind     complement(6122..6139)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6128..6352
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6426..6445)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6427..6445)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6452..6469)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6452..6468)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6461..6483)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6610..7065
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6697..6716)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6907..6928
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7091..7195
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7196..7264,7265..8056)
                     /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(7414..7433)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8227..8815
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8716..8735
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8969..8986
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9103..9124
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9139..9144,9145..9162,9163..9169)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9177..9193
                     /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     9182..9204
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9201..9217
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9201..9217
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        9238..9256
                     /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 gacagcttcc ttactggccg 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 agatgactag ggaggtcagg
     2461 gtcaggagcc cccccctgaa cccaggataa ccctcaaagt cggggggcaa cccgtcacct
     2521 tcctggtaga tactggggcc caacactccg tgctgaccca aaatcctgga cccctaagtg
     2581 ataagtctgc ctgggtccaa ggggctactg gaggaaagcg gtatcgctgg accacggatc
     2641 gcaaagtaca tctagctacc ggtaaggtca cccactcttt cctccatgta ccagactgtc
     2701 cctatcctct gttaggaaga gatttgctga ctaaactaaa agcccaaatc cactttgagg
     2761 gatcaggagc tcaggttatg ggaccaatgg ggcagcccct gcaagtgttg accctaaata
     2821 tagaagatga gtatcggcta catgagacct caaaagagcc agatgtttct ctagggtcca
     2881 catggctgtc tgattttcct caggcctggg cggaaaccgg gggcatggga ctggcagttc
     2941 gccaagctcc tctgatcata cctctgaaag caacctctac ccccgtgtcc ataaaacaat
     3001 accccatgtc acaagaagcc agactgggga tcaagcccca catacagaga ctgttggacc
     3061 agggaatact ggtaccctgc cagtccccct ggaacacgcc cctgctaccc gttaagaaac
     3121 cagggactaa tgattatagg cctgtccagg atctgagaga agtcaacaag cgggtggaag
     3181 acatccaccc caccgtgccc aacccttaca acctcttgag cgggctccca ccgtcccacc
     3241 agtggtacac tgtgcttgat ttaaaggatg cctttttctg cctgagactc caccccacca
     3301 gtcagcctct cttcgccttt gagtggagag atccagagat gggaatctca ggacaattga
     3361 cctggaccag actcccacag ggtttcaaaa acagtcccac cctgtttgat gaggcactgc
     3421 acagagacct agcagacttc cggatccagc acccagactt gatcctgcta cagtacgtgg
     3481 atgacttact gctggccgcc acttctgagc tagactgcca acaaggtact cgggccctgt
     3541 tacaaaccct agggaacctc gggtatcggg cctcggccaa gaaagcccaa atttgccaga
     3601 aacaggtcaa gtatctgggg tatcttctaa aagagggtca gagatggctg actgaggcca
     3661 gaaaagagac tgtgatgggg cagcctactc cgaagacccc tcgacaacta agggagttcc
     3721 tagggacggc aggcttctgt cgcctctgga tccctgggtt tgcagaaatg gcagccccct
     3781 tgtaccctct caccaaaacg gggactctgt ttaattgggg cccagaccaa caaaaggcct
     3841 atcaagaaat caagcaagct cttctaactg ccccagccct ggggttgcca gatttgacta
     3901 agccctttga actctttgtc gacgagaagc agggctacgc caaaggtgtc ctaacgcaaa
     3961 aactgggacc ttggcgtcgg ccggtggcct acctgtccaa aaagctagac ccagtagcag
     4021 ctgggtggcc cccttgccta cggatggtag cagccattgc cgtactgaca aaggatgcag
     4081 gcaagctaac catgggacag ccactagtca ttctggcccc ccatgcagta gaggcactag
     4141 tcaaacaacc ccccgaccgc tggctttcca acgcccggat gactcactat caggccttgc
     4201 ttttggacac ggaccgggtc cagttcggac cggtggtagc cctgaacccg gctacgctgc
     4261 tcccactgcc tgaggaaggg ctgcaacaca actgccttga tatcctggcc gaagcccacg
     4321 gaacccgacc cgacctaacg gaccagccgc tcccagacgc cgaccacacc tggtacacgg
     4381 atggaagcag tctcttacaa gagggacagc gtaaggcggg agctgcggtg accaccgaga
     4441 ccgaggtaat ctgggctaaa gccctgccag ccgggacatc cgctcagcgg gctgaactga
     4501 tagcactcac ccaggcccta aagatggcag aaggtaagaa gctaaatgtt tatactgata
     4561 gccgttatgc ttttgctact gcccatatcc atggagaaat atacagaagg cgtgggttgc
     4621 tcacatcaga aggcaaagag atcaaaaata aagacgagat cttggcccta ctaaaagccc
     4681 tctttctgcc caaaagactt agcataatcc attgtccagg acatcaaaag ggacacagcg
     4741 ccgaggctag aggcaaccgg atggctgacc aagcggcccg aaaggcagcc atcacagaga
     4801 ctccagacac ctctaccctc ctcatagaaa attcatcacc ctacacctca gaacattttc
     4861 attacacagt gactgatata aaggacctaa ccaagttggg ggccatttat gataaaacaa
     4921 agaagtattg ggtctaccaa ggaaaacctg tgatgcctga ccagtttact tttgaattat
     4981 tagactttct tcatcagctg actcacctca gcttctcaaa aatgaaggct ctcctagaga
     5041 gaagccacag tccctactac atgctgaacc gggatcgaac actcaaaaat atcactgaga
     5101 cctgcaaagc ttgtgcacaa gtcaacgcca gcaagtctgc cgttaaacag ggaactaggg
     5161 tccgcgggca tcggcccggc actcattggg agatcgattt caccgagata aagcccggat
     5221 tgtatggcta taaatatctt ctagttttta tagatacctt ttctggctgg atagaagcct
     5281 tcccaaccaa gaaagaaacc gccaaggtcg taaccaagaa gctactagag gagatcttcc
     5341 ccaggttcgg catgcctcag gtattgggaa ctgacaatgg gcctgccttc gtctccaagg
     5401 tgagtcagac agtggccgat ctgttgggga ttgattggaa attacattgt gcatacagac
     5461 cccaaagctc aggccaggta gaaagaatga atagaaccat caaggagact ttaactaaat
     5521 taacgcttgc aactggctct agagactggg tgctcctact ccccttagcc ctgtaccgag
     5581 cccgcaacac gccgggcccc catggcctca ccccatatga gatcttatat ggggcacccc
     5641 cgccccttgt aaacttccct gaccctgaca tgacaagagt tactaacagc ccctctctcc
     5701 aagctcactt acaggctctc tacttagtcc agcacgaagt ctggagacct ctggcggcag
     5761 cctaccaaga acaactggac cgaccggtgg tacctcaccc ttaccgagtc ggcgacacag
     5821 tgtgggtccg ccgacaccag actaagaacc tagaacctcg ctggaaagga ccttacacag
     5881 tcctgctgac cacccccacc gccctcaaag tagacggcat cgcagcttgg atacacgccg
     5941 cccacgtgaa ggctgccgac cccgggggtg gaccatcctc tagactgaca tggcgcgttc
     6001 aacgctctca aaacccctta aaaataaggt tgacgcgtca acccgcgagg ccccctaatc
     6061 cccttaattc ttctgatgct cagaggggtc agtatcgaat tcctgcagcc ccgctgatca
     6121 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     6181 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     6241 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     6301 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     6361 gcggaaagaa ccagctgggg ctcgatcctc tagagcggcc gccaccgcgg tggagctcca
     6421 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
     6481 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     6541 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     6601 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     6661 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     6721 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     6781 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     6841 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     6901 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     6961 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     7021 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     7081 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     7141 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     7201 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     7261 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     7321 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     7381 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     7441 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     7501 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     7561 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     7621 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     7681 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     7741 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     7801 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     7861 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg gttcccgcgg
     7921 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     7981 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     8041 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     8101 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     8161 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     8221 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     8281 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     8341 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     8401 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     8461 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     8521 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     8581 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     8641 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     8701 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     8761 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     8821 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     8881 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     8941 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     9001 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     9061 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     9121 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     9181 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agctcgaaat
     9241 taaccctcac taaagggaac aaaagctggt accgggcccc ccctcgagat ccggccatta
     9301 gccatattat tcattggtta tatagc
//
