LOCUS       sequence-384690-        8881 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MMLV-Gag fused to hMLH1dn with TSTLLMENSS cleavage site between 
            Gag-NES and hMLH1dn. For production of virus like particles with 
            base-editor RNP cargo..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8881)
  AUTHORS   Yu G, Kim HK, Park J, Kwak H, Cheong Y, Kim D, Kim J, Kim J, Kim HH
  TITLE     Prediction of efficiencies for diverse prime editing systems in 
            multiple cell types.
  JOURNAL   Cell. 2023 Apr 21:S0092-8674(23)00331-8. doi: 
            10.1016/j.cell.2023.03.034.
  PUBMED    37119812
REFERENCE   2  (bases 1 to 8881)
  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: "Cell. 2023 
            Apr 21:S0092-8674(23)00331-8. doi: 10.1016/j.cell.2023.03.034."
COMMENT     Sequence Label: pCMV-MMLV-gag-hMLH1dn
FEATURES             Location/Qualifiers
     source          1..8881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(22..41)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(701..1492,1493..1561))
                     /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     1324..1343
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1562..1666)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1692..2147)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1829..1850)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2041..2060
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2274..2296
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2288..2305
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2289..2305
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2315..2334
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2315..2333
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        2454..2833
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2834..3037
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2988..3008
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3034..3058
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          3149..3721
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             3819..4235
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4165..4187
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4203..4219
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             5448..5474
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             5487..5513
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             5526..5552
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             5586..5609
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5643..5663
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(7943..7962)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    8008..8063
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(8062..8081)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(8421..8437)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8421..8437)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8434..8456)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8445..8461
                     /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)."
     promoter        complement(join(8469..8475,8476..8493,8494..8499))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8514..8535
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8652..8669)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(8823..8881,1..530))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
       61 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      121 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      181 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      241 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      301 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      361 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      421 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      481 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      541 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      601 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      661 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      721 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
      781 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
      841 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
      901 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
      961 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1021 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1081 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1141 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1201 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1261 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1321 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1381 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1441 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1501 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1561 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1621 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1681 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1741 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     1801 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     1861 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     1921 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     1981 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2041 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2101 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2161 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2221 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     2281 acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt
     2341 ggagctccac cgcggtggcg gccgctctag agagcttggc ccattgcata cgttgtatcc
     2401 atatcataat atgtacattt atattggctc atgtccaaca ttaccgccat gttgacattg
     2461 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     2521 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     2581 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     2641 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     2701 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     2761 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     2821 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     2881 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     2941 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     3001 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc
     3061 ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct
     3121 ccggtcgacc gatcctgaga acttcagggt gagtttgggg acccttgatt gttctttctt
     3181 tttcgctatt gtaaaattca tgttatatgg agggggcaaa gttttcaggg tgttgtttag
     3241 aatgggaaga tgtcccttgt atcaccatgg accctcatga taattttgtt tctttcactt
     3301 tctactctgt tgacaaccat tgtctcctct tattttcttt tcattttctg taactttttc
     3361 gttaaacttt agcttgcatt tgtaacgaat ttttaaattc acttttgttt atttgtcaga
     3421 ttgtaagtac tttctctaat cacttttttt tcaaggcaat cagggtatat tatattgtac
     3481 ttcagcacag ttttagagaa caattgttat aattaaatga taaggtagaa tatttctgca
     3541 tataaattct ggctggcgtg gaaatattct tattggtaga aacaactaca ccctggtcat
     3601 catcctgcct ttctctttat ggttacaatg atatacactg tttgagatga ggataaaata
     3661 ctctgagtcc aaaccgggcc cctctgctaa ccatgttcat gccttcttct ctttcctaca
     3721 gctcctgggc aacgtgctgg ttgttgtgct gtctcatcat tttggcaaag aattcctcga
     3781 gggccggatc cgaattcctg tatttgtctg aaaacatggg ccagactgtt accactccct
     3841 taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag
     3901 atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg
     3961 gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct
     4021 tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct
     4081 tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc
     4141 ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct
     4201 ccctttatcc agccctcact ccttctctag gcgccaaacc taaacctcaa gttctttctg
     4261 acagtggggg gccgctcatc gacctactta cagaagaccc cccgccttat agggacccaa
     4321 gaccaccccc ttccgacagg gacggaaatg gtggagaagc gacccctgcg ggagaggcac
     4381 cggacccctc cccaatggca tctcgcctac gtgggagacg ggagccccct gtggccgact
     4441 ccactacctc gcaggcattc cccctccgcg caggaggaaa cggacagctt caatactggc
     4501 cgttctcctc ttctgacctt tacaactgga aaaataataa cccttctttt tctgaagatc
     4561 caggtaaact gacagctctg atcgagtctg tcctcatcac ccatcagccc acctgggacg
     4621 actgtcagca gctgttgggg actctgctga ccggagaaga aaaacaacgg gtgctcttag
     4681 aggctagaaa ggcggtgcgg ggcgatgatg ggcgccccac tcaactgccc aatgaagtcg
     4741 atgccgcttt tcccctcgag cgcccagact gggattacac cacccaggca ggtaggaacc
     4801 acctagtcca ctatcgccag ttgctcctag cgggtctcca aaacgcgggc agaagcccca
     4861 ccaatttggc caaggtaaaa ggaataacac aagggcccaa tgagtctccc tcggccttcc
     4921 tagagagact taaggaagcc tatcgcaggt acactcctta tgaccctgag gacccagggc
     4981 aagaaactaa tgtgtctatg tctttcattt ggcagtctgc cccagacatt gggagaaagt
     5041 tagagaggtt agaagattta aaaaacaaga cgcttggaga tttggttaga gaggcagaaa
     5101 agatctttaa taaacgagaa accccggaag aaagagagga acgtatcagg agagaaacag
     5161 aggaaaaaga agaacgccgt aggacagagg atgagcagaa agagaaagaa agagatcgta
     5221 ggagacatag agagatgagc aagctattgg ccactgtcgt tagtggacag aaacaggata
     5281 gacagggagg agaacgaagg aggtcccaac tcgatcgcga ccagtgtgcc tactgcaaag
     5341 aaaaggggca ctgggctaaa gattgtccca agaaaccacg aggacctcgg ggaccaagac
     5401 cccagacctc cctcctgacc ctagatgact ctggcggctc acttcaactg cctccacttg
     5461 aaagactgac actgggatca ttacaattac ctcctttaga acgattaaca ctcggttcac
     5521 tacagcttcc gcctcttgag agattgacat taacgtccac gctgctaatg gagaactcgt
     5581 ctggagatta caaagacgat gacgataaga aacgtacggc cgacggaagc gagttcgagt
     5641 caccaaagaa gaagcggaaa gtcagcgggg ggtcaagctt cgttgctgga gtcatccgga
     5701 gactggacga gacagtggtg aacagaattg ccgccggcga ggtgatccag agacctgcca
     5761 atgcaatcaa ggagatgatc gagaactgtc tggacgccaa gtccacaagc attcaggtga
     5821 tcgtgaagga gggcggactg aagctgatcc agatccaaga caacggcaca ggcatcagaa
     5881 aggaagatct ggacatcgtg tgtgaacggt tcaccacatc taagctgcag tcttttgagg
     5941 atctggcctc tatcagtacc tacggcttca gaggcgaggc cctggccagc atcagccacg
     6001 tggcccatgt gaccatcacc accaaaaccg ccgacggcaa atgcgcttat cgcgctagct
     6061 acagcgacgg caagctgaaa gccccgccaa agccttgcgc cggcaaccag ggtacacaga
     6121 taacagtgga ggatctgttc tacaacatcg ccacccggag aaaggccctg aaaaatccca
     6181 gcgaggagta cggcaagatc ctggaagtcg tcggcagata ctccgtgcac aacgccggaa
     6241 tcagctttag cgtaaagaag cagggagaaa ccgtggccga tgtgcgcacc ctgccaaatg
     6301 ccagcaccgt ggataacatc agaagcattt tcggaaatgc cgtgtccaga gaactgatcg
     6361 agatcggctg cgaagataag accctggctt ttaagatgaa cggctacatc agcaacgcca
     6421 attactctgt gaagaagtgc atctttcttc tgttcatcaa ccacagactg gtggaaagca
     6481 ccagcctgcg gaaagccatc gagacagtgt acgccgccta cctgcctaag aacacccacc
     6541 ccttcctgta cctgagcctc gagatcagcc ctcagaacgt ggacgtcaat gtgcatccta
     6601 caaagcacga ggtgcacttc ctgcacgagg aaagcatcct ggaaagagtg cagcagcaca
     6661 ttgagagcaa gctgctgggc tctaacagca gcagaatgta cttcacacag accctgctgc
     6721 ctggcctggc cggcccctca ggcgaaatgg ttaagtccac aacctctctg acctcatcca
     6781 gcaccagcgg ttcttccgat aaggtgtacg cccaccagat ggtgcggacc gactctcggg
     6841 agcagaagct ggacgccttt ctgcaacctc tgagcaaacc tctgagctct cagcctcagg
     6901 ccatcgtgac cgaggacaag acagatatct cctccggccg tgccagacag caggacgaag
     6961 aaatgctcga gctgccagct cctgccgagg tggccgccaa gaaccagagc ctggagggag
     7021 ataccacaaa gggcaccagc gaaatgagcg agaagcgggg ccctacctct agcaacccca
     7081 gaaaacggca ccgggaggac agcgacgtgg aaatggtgga ggacgacagc agaaaggaaa
     7141 tgacagccgc ttgtacccct agaagaagaa tcatcaacct gacctccgtg ctgagcctgc
     7201 aggaggagat caacgagcag ggccacgagg tgctgagaga gatgctgcac aatcacagct
     7261 tcgtgggctg cgtgaaccct caatgggccc tggctcagca tcaaacaaag ctgtacctgc
     7321 tgaacaccac caagctgagc gaagagctgt tctaccagat cctcatctac gacttcgcca
     7381 acttcggcgt gctacgcctg agcgagcccg cccctctgtt tgacctggcc atgctggctc
     7441 tggatagccc agaaagcggc tggacagaag aggacggacc taaagagggg ctggctgaat
     7501 acatcgtgga gttcctgaag aaaaaggccg agatgctggc cgactacttt tctctggaaa
     7561 tcgacgagga aggcaacctg atcggcctgc ctctgctgat cgataactac gtgcctcccc
     7621 tggaaggcct gcccatcttc atcctgagac tggctacaga ggtgaactgg gacgaggaaa
     7681 aggaatgctt cgagtctctg agcaaggagt gcgccatgtt ctatagcatc agaaaacagt
     7741 acatctctga agagagcact ctgtctggcc agcagagtga agtgcccgga agcatcccca
     7801 acagctggaa gtggaccgtg gaacacatcg tgtacaaggc cctgcggagc cacattctcc
     7861 ctcctaagca cttcaccgag gacggcaaca tcctgcagct ggccaacctg cccgaccttt
     7921 ataaggtttt ctaaaccggt gatctttttc cctctgccaa aaattatggg gacatcatga
     7981 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
     8041 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
     8101 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga
     8161 acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct
     8221 tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat
     8281 ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc
     8341 tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacggat
     8401 cggccgcaat tcgtaaccat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     8461 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     8521 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     8581 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     8641 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     8701 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     8761 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     8821 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     8881 t
//
