LOCUS       sequence-407420-        7574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Generation of CreVLPs when used in conjunction with MMLV gag-pol 
            during production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7574)
  AUTHORS   Strebinger D, Frangieh CJ, Friedrich MJ, Faure G, Macrae RK, Zhang F
  TITLE     Cell type-specific delivery by modular envelope design.
  JOURNAL   Nat Commun. 2023 Aug 23;14(1):5141. doi: 10.1038/s41467-023-40788-8.
  PUBMED    37612276
REFERENCE   2  (bases 1 to 7574)
  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: "2023-08-23"; volume: "14(1):5141. doi"; pages: " 
            10.1038/s41467-023-40788-8"
COMMENT     Sequence Label: MMLVgag-3NES-Cre
FEATURES             Location/Qualifiers
     source          1..7574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4..420
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     350..372
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     388..404
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1633..1659
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             1672..1698
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             1711..1737
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             1771..1791
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1792..2820
                     /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="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLQ
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     primer_bind     complement(2821..2840)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2886..2941
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2940..2959)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3299..3315)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3299..3315)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3312..3334)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3323..3339
                     /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(3347..3353,3354..3371,3372..3377))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3392..3413
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3530..3547)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3701..4289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3781..3800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4460..5251,5252..5320))
                     /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     5083..5102
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5321..5425)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5451..5906)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5588..5609)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5800..5819
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6033..6055
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6047..6064
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6048..6064
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6074..6093
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6074..6092
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        6213..6592
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6593..6796
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6747..6767
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6793..6817
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          6908..7480
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
ORIGIN
        1 atgggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa agatgtcgag
       61 cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac cttctgctct
      121 gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc
      181 atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc agaccaggtc
      241 ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt caagcccttt
      301 gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc ccttgaacct
      361 cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc tctaggcgcc
      421 aaacctaaac ctcaagttct ttctgacagt ggggggccgc tcatcgacct acttacagaa
      481 gaccccccgc cttataggga cccaagacca cccccttccg acagggacgg aaatggtgga
      541 gaagcgaccc ctgcgggaga ggcaccggac ccctccccaa tggcatctcg cctacgtggg
      601 agacgggagc cccctgtggc cgactccact acctcgcagg cattccccct ccgcgcagga
      661 ggaaacggac agcttcaata ctggccgttc tcctcttctg acctttacaa ctggaaaaat
      721 aataaccctt ctttttctga agatccaggt aaactgacag ctctgatcga gtctgtcctc
      781 atcacccatc agcccacctg ggacgactgt cagcagctgt tggggactct gctgaccgga
      841 gaagaaaaac aacgggtgct cttagaggct agaaaggcgg tgcggggcga tgatgggcgc
      901 cccactcaac tgcccaatga agtcgatgcc gcttttcccc tcgagcgccc agactgggat
      961 tacaccaccc aggcaggtag gaaccaccta gtccactatc gccagttgct cctagcgggt
     1021 ctccaaaacg cgggcagaag ccccaccaat ttggccaagg taaaaggaat aacacaaggg
     1081 cccaatgagt ctccctcggc cttcctagag agacttaagg aagcctatcg caggtacact
     1141 ccttatgacc ctgaggaccc agggcaagaa actaatgtgt ctatgtcttt catttggcag
     1201 tctgccccag acattgggag aaagttagag aggttagaag atttaaaaaa caagacgctt
     1261 ggagatttgg ttagagaggc agaaaagatc tttaataaac gagaaacccc ggaagaaaga
     1321 gaggaacgta tcaggagaga aacagaggaa aaagaagaac gccgtaggac agaggatgag
     1381 cagaaagaga aagaaagaga tcgtaggaga catagagaga tgagcaagct attggccact
     1441 gtcgttagtg gacagaaaca ggatagacag ggaggagaac gaaggaggtc ccaactcgat
     1501 cgcgaccagt gtgcctactg caaagaaaag gggcactggg ctaaagattg tcccaagaaa
     1561 ccacgaggac ctcggggacc aagaccccag acctccctcc tgaccctaga tgactctggc
     1621 ggctcacttc aactgcctcc acttgaaaga ctgacactgg gatcattaca attacctcct
     1681 ttagaacgat taacactcgg ttcactacag cttccgcctc ttgagagatt gacattaacg
     1741 tccacgctgc taatggagaa ctcgtctgga cccaagaaaa agcgcaaggt atccaattta
     1801 ctgaccgtac accaaaattt gcctgcatta ccggtcgatg caacgagtga tgaggttcgc
     1861 aagaacctga tggacatgtt cagggatcgc caggcgtttt ctgagcatac ctggaaaatg
     1921 cttcagtccg tttgccggtc gtgggcggca tggtgcaagt tgaataaccg gaaatggttt
     1981 cccgcagaac ctgaagatgt tcgcgattat cttctatatc ttcaggcgcg cggtctggca
     2041 gtaaaaacta tccagcaaca tttgggccag ctaaacatgc ttcatcgtcg gtccgggctg
     2101 ccacgaccaa gtgacagcaa tgctgtttca ctggttatgc ggcggatccg aaaagaaaac
     2161 gttgatgccg gtgaacgtgc aaaacaggct ctagcgttcg aacgcactga tttcgaccag
     2221 gttcgttcac tcatggaaaa tagcgatcgc tgccaggata tacgtaatct ggcatttctg
     2281 gggattgctt ataacaccct gttacgtata gccgaaattg ccaggatcag ggttaaagat
     2341 atctcacgta ctgacggtgg gagaatgtta atccatattg gcagaacgaa aacgctggtt
     2401 agcaccgcag gtgtagagaa ggcacttagc ctgggggtaa ctaaactggt cgagcgatgg
     2461 atttccgtct ctggtgtagc tgatgatccg aataactacc tgttttgccg ggtcagaaaa
     2521 aatggtgttg ccgcgccatc tgccaccagc cagctatcaa ctcgcgccct ggaagggatt
     2581 tttgaagcaa ctcatcgatt gatttacggc gctaaggatg actctggtca gagatacctg
     2641 gcctggtctg gacacagtgc ccgtgtcgga gccgcgcgag atatggcccg cgctggagtt
     2701 tcaataccgg agatcatgca agctggtggc tggaccaatg taaatattgt catgaactat
     2761 atccgtaacc tggatagtga aacaggggca atggtgcgcc tgctggaaga tggcgattga
     2821 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
     2881 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     2941 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
     3001 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg
     3061 tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac
     3121 ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa
     3181 attttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat
     3241 agctgtccct cttctcttat ggagatccct cgacggatcg gccgcaattc gtaaccatgt
     3301 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     3361 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     3421 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3481 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3541 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3601 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3661 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3721 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3781 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3841 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3901 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3961 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4021 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4081 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4141 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4201 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4261 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4321 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4381 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4441 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4501 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4561 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4621 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4681 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4741 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4801 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4861 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4921 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4981 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5041 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5101 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5161 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5221 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5281 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5341 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5401 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag
     5461 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     5521 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     5581 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     5641 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     5701 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     5761 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     5821 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     5881 gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga
     5941 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     6001 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     6061 cagtgagcgc gcgtaatacg actcactata gggcgaattg gagctccacc gcggtggcgg
     6121 ccgctctaga gagcttggcc cattgcatac gttgtatcca tatcataata tgtacattta
     6181 tattggctca tgtccaacat taccgccatg ttgacattga ttattgacta gttattaata
     6241 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     6301 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     6361 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     6421 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     6481 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     6541 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     6601 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     6661 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     6721 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     6781 ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg
     6841 ttttgacctc catagaagac accgggaccg atccagcctc cggtcgaccg atcctgagaa
     6901 cttcagggtg agtttgggga cccttgattg ttctttcttt ttcgctattg taaaattcat
     6961 gttatatgga gggggcaaag ttttcagggt gttgtttaga atgggaagat gtcccttgta
     7021 tcaccatgga ccctcatgat aattttgttt ctttcacttt ctactctgtt gacaaccatt
     7081 gtctcctctt attttctttt cattttctgt aactttttcg ttaaacttta gcttgcattt
     7141 gtaacgaatt tttaaattca cttttgttta tttgtcagat tgtaagtact ttctctaatc
     7201 actttttttt caaggcaatc agggtatatt atattgtact tcagcacagt tttagagaac
     7261 aattgttata attaaatgat aaggtagaat atttctgcat ataaattctg gctggcgtgg
     7321 aaatattctt attggtagaa acaactacac cctggtcatc atcctgcctt tctctttatg
     7381 gttacaatga tatacactgt ttgagatgag gataaaatac tctgagtcca aaccgggccc
     7441 ctctgctaac catgttcatg ccttcttctc tttcctacag ctcctgggca acgtgctggt
     7501 tgttgtgctg tctcatcatt ttggcaaaga attcctcgag ggccggatcc gaattcctgt
     7561 atttgtctga aaac
//
