LOCUS       sequence-462772-        5642 bp    DNA     circular SYN 15-APR-2025
DEFINITION  expression plasmid to produce Cre-NLS 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 5642)
  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, Rodrigues Menezes C, Risaliti E, Engfer ZJ, Koleci N, 
            Adrea Schmidts A, Geerlof A, Palczewski K, Westmeyer GG, Truong DJ
  TITLE     Engineered nucleocytosolic vehicles for loading of programmable 
            editors
  JOURNAL   Cell, April 2025. doi: 10.1016/j.cell.2025.03.015.
REFERENCE   2  (bases 1 to 5642)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell, April
            2025. doi: 10.1016/j.cell.2025.03.015."
COMMENT     Sequence Label: pCMV_CreNLS_PP7
FEATURES             Location/Qualifiers
     source          1..5642
                     /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"
     CDS             2411..2437
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     primer_bind     complement(2540..2556)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2570..2587)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2576..2800
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2872..2889)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2872..2888)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2881..2903)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3029..3484)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3166..3187)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3378..3397
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3511..3615
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3616..3684,3685..4476)
                     /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(3834..3853)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4647..5235
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5136..5155
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5389..5406
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5523..5544
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5559..5564,5565..5582,5583..5589)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5597..5613
                     /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     5602..5624
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5621..5637
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5621..5637
                     /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 tgagcaacct
     1381 gctgaccgtg caccagaacc tgcctgctct gcctgtggac gccacatctg atgaagtgcg
     1441 gaagaacctg atggacatgt tcagagacag acaggccttc agcgagcaca cctggaagat
     1501 gctgctgagc gtgtgtagaa gctgggccgc ttggtgcaag ctgaacaaca gaaagtggtt
     1561 ccccgccgag cctgaggacg tgcgagatta cctgctgtac ctgcaagcta gaggcctggc
     1621 cgtgaaaacc atccagcagc acctgggcca gctgaacatg ctgcacagaa gaagcggcct
     1681 gcctagacct agcgacagca acgctgtgtc cctggtcatg agaaggattc ggaaagaaaa
     1741 cgtggacgct ggcgagagag ctaagcaggc tctggccttc gagagaaccg acttcgatca
     1801 agtgcgcagc ctgatggaaa acagcgacag atgccaggat attcggaacc tggccttcct
     1861 gggaatcgcc tacaacaccc tgctgagaat cgccgagatc gccagaatca gagtgaagga
     1921 catcagcaga accgacggcg gcagaatgct gatccacatc ggcagaacaa agaccctggt
     1981 gtccacagct ggcgtcgaga aggctctgag tctgggcgtg acaaagctgg tggaaagatg
     2041 gatcagcgtg tccggcgtgg ccgacgatcc taacaactac ctgttctgtc gcgtgcgcaa
     2101 gaacggcgtg gcagctcctt ctgctacaag ccagctgagc acaagagccc tggaaggcat
     2161 cttcgaggcc acacacagac tgatctacgg cgccaaggat gacagcggcc agagatacct
     2221 tgcttggagc ggccacagtg ctagagtggg cgctgctaga gacatggcta gagcaggcgt
     2281 gtcaatcccc gagatcatgc aagctggcgg ctggaccaac gtgaacatcg tgatgaacta
     2341 catccgcaac ctggacagcg agacaggcgc tatggttcga ctgcttgaag atggcgacgg
     2401 tggatccggt cctgccgcta agagagtgaa gctggactga acgcgtgctc gctttcttgc
     2461 tgtccaattt ctattaaagg ttcctttgtt cgcgttgctg gaccagaaga tatggcttcg
     2521 gttcagcaaa atgattgcag atccactagt tctagagctc gctgatcagc ctcgactgtg
     2581 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     2641 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     2701 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     2761 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     2821 agctggggct cgagatccac tagttctagc ctcgaggcta gagcggccgc cactggccgt
     2881 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     2941 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     3001 acagttgcgc agcctgaatg gcgaatggaa attgtaagcg ttaatatttt gttaaaattc
     3061 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     3121 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     3181 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     3241 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     3301 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     3361 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     3421 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     3481 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3541 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3601 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     3661 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3721 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3781 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     3841 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     3901 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     3961 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4021 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4081 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     4141 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     4201 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     4261 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     4321 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     4381 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4441 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     4501 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4561 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4621 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4681 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4741 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4801 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4861 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4921 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4981 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5041 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5101 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5161 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5221 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5281 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5341 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     5401 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     5461 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     5521 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     5581 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     5641 ga
//
