LOCUS       sequence-234838-        7529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GAG (FMLV) fused with CRE recombinase for the production 
            of VLPs loaded with CRE protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7529)
  AUTHORS   Mangeot PE, Risson V, Fusil F, Marnef A, Laurent E, Blin J, 
            Mournetas V, Massourides E, Sohier TJM, Corbin A, Aube F, Teixeira 
            M, Pinset C, Schaeffer L, Legube G, Cosset FL, Verhoeyen E, Ohlmann 
            T, Ricci EP
  TITLE     Genome editing in primary cells and in vivo using viral-derived 
            Nanoblades loaded with Cas9-sgRNA ribonucleoproteins.
  JOURNAL   Nat Commun. 2019 Jan 3;10(1):45. doi: 10.1038/s41467-018-07845-z.
  PUBMED    30604748
REFERENCE   2  (bases 1 to 7529)
  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 
            Commun."; date: "2019-01-3"; volume: "10(1):45. doi"; pages: " 
            10.1038/s41467-018-07845-z"
COMMENT     Sequence Label: GAG-CRErec
FEATURES             Location/Qualifiers
     source          1..7529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..31
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     23..40
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     24..40
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     50..69
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        50..68
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        189..568
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        569..772
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     723..743
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     769..793
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          884..1456
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             3192..3212
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3213..4241
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     primer_bind     complement(4326..4345)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4391..4446
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4445..4464)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4804..4820)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4804..4820)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4817..4839)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4828..4844
                     /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(4852..4858,4859..4876,4877..4882))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4897..4918
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5035..5052)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5206..5794)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5286..5305)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5965..6756,6757..6825))
                     /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     6588..6607
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6826..6930)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6956..7411)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7093..7114)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7305..7324
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc
       61 actatagggc gaattggagc tccaccgcgg tggcggccgc tctagagagc ttggcccatt
      121 gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc caacattacc
      181 gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
      241 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      301 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      361 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      421 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      481 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      541 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      601 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      661 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      721 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
      781 gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg
      841 ggaccgatcc agcctccggt cgaccgatcc tgagaacttc agggtgagtt tggggaccct
      901 tgattgttct ttctttttcg ctattgtaaa attcatgtta tatggagggg gcaaagtttt
      961 cagggtgttg tttagaatgg gaagatgtcc cttgtatcac catggaccct catgataatt
     1021 ttgtttcttt cactttctac tctgttgaca accattgtct cctcttattt tcttttcatt
     1081 ttctgtaact ttttcgttaa actttagctt gcatttgtaa cgaattttta aattcacttt
     1141 tgtttatttg tcagattgta agtactttct ctaatcactt ttttttcaag gcaatcaggg
     1201 tatattatat tgtacttcag cacagtttta gagaacaatt gttataatta aatgataagg
     1261 tagaatattt ctgcatataa attctggctg gcgtggaaat attcttattg gtagaaacaa
     1321 ctacaccctg gtcatcatcc tgcctttctc tttatggtta caatgatata cactgtttga
     1381 gatgaggata aaatactctg agtccaaacc gggcccctct gctaaccatg ttcatgcctt
     1441 cttctctttc ctacagctcc tgggcaacgt gctggttgtt gtgctgtctc atcattttgg
     1501 caaagaattc ctcgagggcc ggatccgaat tcctgtattt gtctgaaaac atgggccagg
     1561 ctgttaccac ccccttaagt ttgactttag accactggaa ggatgtcgaa cggacagccc
     1621 acaacctgtc ggtagaggtt agaaaaaggc gctgggttac attctgctct gcagaatggc
     1681 caaccttcaa cgtcggatgg ccacgagacg gcacttttaa cccagacatt attacacagg
     1741 ttaagatcaa ggtcttctca cctggcccac atggacatcc ggatcaggtc ccctacatcg
     1801 tgacctggga agctatagca gtagaccccc ctccctgggt cagacccttc gtgcacccta
     1861 aacctcccct ctctcttccc ccttcagccc cctctctccc acctgaaccc ccactctcga
     1921 ccccgcccca gtcctccctc tatccggctc tcacttctcc tttaaacacc aaacctaggc
     1981 ctcaagtcct tcctgatagc ggaggaccac tcattgatct actcacggag gaccctccgc
     2041 cttaccggga cccagggcca ccctctcctg acgggaacgg cgatagcgga gaagtggccc
     2101 ctacagaagg agcccctgac ccttccccaa tggtatcccg cctgcgggga agaaaagaac
     2161 cccccgtggc ggattctact acctctcagg cgttccccct tcgcctggga gggaatggac
     2221 agtatcaata ctggccattt tcctcctctg acctctataa ctggaaaaat aacaacccct
     2281 ctttctccga ggacccagct aaattgacag ctttgatcga gtccgttctc cttactcatc
     2341 agcccacttg ggatgactgc caacagctat tagggaccct gctgacggga gaagaaaaac
     2401 agcgagtgct cctagaggcc cgaaaggcgg ttcgagggga ggacggacgc ccaactcagc
     2461 tgcccaatga cattaatgat gcttttccct tggaacgtcc cgactgggac tacaacaccc
     2521 aacgaggtag gaaccaccta gtccactatc gccagttgct cctagcgggt ctccaaaacg
     2581 cgggcagaag ccccaccaat ttggccaagg taaaagggat aacccaggga cctaatgagt
     2641 ctccctcagc ctttttagag agactcaagg aggcctatcg cagatacact ccttatgacc
     2701 ctgaggaccc agggcaagaa accaatgtgg ccatgtcatt catctggcag tccgccccgg
     2761 atatcgggcg aaagttagag cggttagaag atttgaagag taagacctta ggagacttag
     2821 tgagggaagc tgaaaagatc tttaataaac gagaaacccc ggaagaaaga gaggaacgta
     2881 ttaggagaga aacagaggaa aaggaagaac gccgtagggc agaggatgtg cagagagaga
     2941 aggagaggga ccgcagaaga catagagaaa tgagtaagtt gctggctact gtcgttagcg
     3001 ggcagagaca ggatagacag ggaggagagc gaaggaggcc ccaactcgac cacgaccagt
     3061 gtgcctactg caaagaaaag ggacattggg ctagagattg ccccaagaag ccaagaggac
     3121 cccggggacc acgaccccag gcctccctcc tgacgcgtag ttccctgtat ccagccctca
     3181 cacctaccgg ccccaagaag aagaggaagg tgtccaattt actgaccgta caccaaaatt
     3241 tgcctgcatt accggtcgat gcaacgagtg atgaggttcg caagaacctg atggacatgt
     3301 tcagggatcg ccaggcgttt tctgagcata cctggaaaat gcttctgtcc gtttgccggt
     3361 cgtgggcggc atggtgcaag ttgaataacc ggaaatggtt tcccgcagaa cctgaagatg
     3421 ttcgcgatta tcttctatat cttcaggcgc gcggtctggc agtaaaaact atccagcaac
     3481 atttgggcca gctaaacatg cttcatcgtc ggtccgggct gccacgacca agtgacagca
     3541 atgctgtttc actggttatg cggcggatcc gaaaagaaaa cgttgatgcc ggtgaacgtg
     3601 caaaacaggc tctagcgttc gaacgcactg atttcgacca ggttcgttca ctcatggaaa
     3661 atagcgatcg ctgccaggat atacgtaatc tggcatttct ggggattgct tataacaccc
     3721 tgttacgtat agccgaaatt gccaggatca gggttaaaga tatctcacgt actgacggtg
     3781 ggagaatgtt aatccatatt ggcagaacga aaacgctggt tagcaccgca ggtgtagaga
     3841 aggcacttag cctgggggta actaaactgg tcgagcgatg gatttccgtc tctggtgtag
     3901 ctgatgatcc gaataactac ctgttttgcc gggtcagaaa aaatggtgtt gccgcgccat
     3961 ctgccaccag ccagctatca actcgcgccc tggaagggat ttttgaagca actcatcgat
     4021 tgatttacgg cgctaaggat gactctggtc agagatacct ggcctggtct ggacacagtg
     4081 cccgtgtcgg agccgcgcga gatatggccc gcgctggagt ttcaataccg gagatcatgc
     4141 aagctggtgg ctggaccaat gtaaatattg tcatgaacta tatccgtaac ctggatagtg
     4201 aaacaggggc aatggtgcgc ctgctggaag atggcgatta ggtacctgca tcctcaggtg
     4261 caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac
     4321 tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     4381 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     4441 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     4501 gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa
     4561 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     4621 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     4681 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     4741 gtcatagctg tccctcttct cttatggaga tccctcgacg gatcggccgc aattcgtaat
     4801 catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4861 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4921 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4981 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5041 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5101 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5161 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5221 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5281 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5341 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5401 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5461 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5521 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5581 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5641 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5701 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5761 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5821 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5881 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5941 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6001 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6061 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6121 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6181 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6241 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6301 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6361 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6421 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6481 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6541 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6601 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6661 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6721 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6781 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6841 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6901 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     6961 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     7021 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     7081 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     7141 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     7201 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     7261 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     7321 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     7381 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     7441 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     7501 gctgcaaggc gattaagttg ggtaacgcc
//
