LOCUS       sequence-462773-        5291 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression plasmid to produce mGreenLantern mRNA with a PP7 aptamer 
            in the 3' UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5291)
  AUTHORS   Geilenkeuser J, Armbrust N, Steinmassl E, Du SW, Schmidt S, Binder 
            EMH, Li Y, Warsing NW, Wendel SV, von der Linde F, Schiele EM, Niu 
            X, Stroppel L, Berezin O, Santl TH, Orschmann T, Nelson K, Gruber C,
            Palczewska G, Menezes CR, Risaliti E, Engfer ZJ, Koleci N, Schmidts 
            A, Geerlof A, Palczewski K, Westmeyer GG, Truong DJ
  TITLE     Engineered nucleocytosolic vehicles for loading of programmable 
            editors.
  JOURNAL   Cell. 2025 May 15;188(10):2637-2655.e31. doi: 
            10.1016/j.cell.2025.03.015. Epub 2025 Apr 9.
  PUBMED    40209705
REFERENCE   2  (bases 1 to 5291)
  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"; doi: 
            "10.1016/j.cell.2025.03.015"; journalName: "Cell"; date: 
            "2025-05-15- 15"; volume: "188"; issue: "10"; pages: "2637-2655.e31"
COMMENT     Sequence Label: pCMV_mGL_PP7
FEATURES             Location/Qualifiers
     source          1..5291
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        19..398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        399..602
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     553..573
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     599..623
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          714..1286
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     1341..1360
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1341..1359
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1364..1373
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1370..1372,1373..1375,1376..2089)
                     /codon_start=1
                     /product="monomeric GFP derivative that displays 
                     exceptionally high brightness in cells (Campbell et al., 
                     2020)"
                     /label=mGreenLantern
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLGYGVACFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIVLKGIDFKEDGNILGHKLEYNFNSHKVYITADKQKNGIK
                     ANFKTRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLK
                     ERVTAAGITHDMDELYK"
     primer_bind     complement(1415..1436)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2189..2205)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2219..2236)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2225..2449
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2521..2538)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2521..2537)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2530..2552)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2678..3133)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2815..2836)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3027..3046
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3160..3264
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3265..3333,3334..4125)
                     /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(3483..3502)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4296..4884
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4785..4804
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5038..5055
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5172..5193
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5208..5213,5214..5231,5232..5238)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5246..5262
                     /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     5251..5273
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5270..5286
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5270..5286
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ggcgcgcccc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
       61 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      121 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      181 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      241 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      301 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      361 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      421 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      481 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      541 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      601 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
      661 gaagacaccg ggaccgatcc agcctccggt cgaccgatcc tgagaacttc agggtgagtt
      721 tggggaccct tgattgttct ttctttttcg ctattgtaaa attcatgtta tatggagggg
      781 gcaaagtttt cagggtgttg tttagaatgg gaagatgtcc cttgtatcac catggaccct
      841 catgataatt ttgtttcttt cactttctac tctgttgaca accattgtct cctcttattt
      901 tcttttcatt ttctgtaact ttttcgttaa actttagctt gcatttgtaa cgaattttta
      961 aattcacttt tgtttatttg tcagattgta agtactttct ctaatcactt ttttttcaag
     1021 gcaatcaggg tatattatat tgtacttcag cacagtttta gagaacaatt gttataatta
     1081 aatgataagg tagaatattt ctgcatataa attctggctg gcgtggaaat attcttattg
     1141 gtagaaacaa ctacaccctg gtcatcatcc tgcctttctc tttatggtta caatgatata
     1201 cactgtttga gatgaggata aaatactctg agtccaaacc gggcccctct gctaaccatg
     1261 ttcatgcctt cttctctttc ctacagctcc tgggcaacgt gctggttgtt gtgctgtctc
     1321 atcattttgg cattaattaa taatacgact cactataggg gccgccacca tggtgagcaa
     1381 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1441 cggccacaag ttcagcgtcc gcggcgaggg cgagggcgat gccaccaacg gcaagctgac
     1501 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1561 cttaggctac ggcgtggcct gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     1621 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatctctt tcaaggacga
     1681 cggcacctac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1741 cgtgctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1801 caacttcaac agccacaagg tctatatcac ggccgacaag cagaagaacg gcatcaaggc
     1861 taacttcaag acccgccaca acgttgagga cggcggcgtg cagctcgccg accactacca
     1921 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcca
     1981 tcagtccaaa ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgaaggagag
     2041 ggtgaccgcc gccgggatta cacatgacat ggacgagctg tacaagtaga cgcgtgctcg
     2101 ctttcttgct gtccaatttc tattaaaggt tcctttgttc gcgttgctgg accagaagat
     2161 atggcttcgg ttcagcaaaa tgattgcaga tccactagtt ctagagctcg ctgatcagcc
     2221 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     2281 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     2341 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2401 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     2461 gaaagaacca gctggggctc gagatccact agttctagcc tcgaggctag agcggccgcc
     2521 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     2581 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     2641 cccttcccaa cagttgcgca gcctgaatgg cgaatggaaa ttgtaagcgt taatattttg
     2701 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     2761 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     2821 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     2881 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     2941 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     3001 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     3061 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     3121 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3181 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3241 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3301 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3361 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3421 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     3481 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     3541 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3601 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3661 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3721 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3781 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     3841 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     3901 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     3961 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     4021 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4081 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4141 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4201 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4261 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4321 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4381 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4441 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4501 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4561 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4621 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4681 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4741 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4801 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4861 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4921 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4981 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     5041 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     5101 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     5161 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     5221 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     5281 tatgaccatg a
//
