LOCUS       sequence-426612-        5177 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Split luciferase assay to test reconstitution via mRNA 
            trans-splicing (REVeRT system)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5177)
  AUTHORS   Riedmayr LM, Hinrichsmeyer KS, Thalhammer SB, Mittas DM, Karguth N, 
            Otify DY, Bohm S, Weber VJ, Bartoschek MD, Splith V, Brummer M, 
            Ferreira R, Boon N, Wogenstein GM, Grimm C, Wijnholds J, Mehlfeld V,
            Michalakis S, Fenske S, Biel M, Becirovic E
  TITLE     mRNA trans-splicing dual AAV vectors for (epi)genome editing and 
            gene therapy.
  JOURNAL   Nat Commun. 2023 Oct 18;14(1):6578. doi: 10.1038/s41467-023-42386-0.
  PUBMED    37852949
REFERENCE   2  (bases 1 to 5177)
  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: "2023-10-18"; volume: "14(1):6578. doi"; pages: " 
            10.1038/s41467-023-42386-0"
COMMENT     Sequence Label: pAAV-CMV-5'Luciferase(split CAGGT)_BDlacZ-SV40...
FEATURES             Location/Qualifiers
     source          1..5177
                     /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"
     regulatory      629..638
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(701..719)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(1812..1831)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    1841..1922
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1866..1885
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2010..2139
                     /label=AAV2 ITR
     primer_bind     complement(2158..2175)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2158..2174)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2167..2189)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2316..2771
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2403..2422)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2613..2634
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2797..2901
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2902..2970,2971..3762)
                     /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(3120..3139)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3933..4521
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4422..4441
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4809..4830
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4845..4850,4851..4868,4869..4875)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4883..4899
                     /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     4888..4910
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4907..4923
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4907..4923
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   4961..5090
                     /label=AAV2 ITR
ORIGIN
        1 aaatagctag cacgcgttga 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 aaccggtcgc caccatggaa gacgccaaaa acataaagaa
      661 aggcccggcg ccattctatc cgctggaaga tggaaccgct ggagagcaac tgcataaggc
      721 tatgaagaga tacgccctgg ttcctggaac aattgctttt acagatgcac atatcgaggt
      781 ggacatcact tacgctgagt acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg
      841 atatgggctg aatacaaatc acagaatcgt cgtatgcagt gaaaactctc ttcaattctt
      901 tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga acgacattta
      961 taatgaacgt gaattgctca acagtatggg catttcgcag cctaccgtgg tgttcgtttc
     1021 caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat
     1081 tattatcatg gattctaaaa cggattacca gggatttcag tcgatgtaca cgttcgtcac
     1141 atctcatcta cctcccggtt ttaatgaata cgattttgtg ccagagtcct tcgataggga
     1201 caagacaatt gcactgatca tgaactcctc tggatctact ggtctgccta aaggtgtcgc
     1261 tctgcctcat agaactgcct gcgtgagatt ctcgcatgcc agagatccta tttttggcaa
     1321 tcaaatcatt ccggatactg cgattttaag tgttgttcca ttccatcacg gttttggaat
     1381 gtttactaca ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt atagatttga
     1441 agaagagctg tttctgagga gccttcagga ttacaagatt caaagtgcgc tgctggtgcc
     1501 aaccctattc tccttcttcg ccaaaagcac tctgattgac aaatacgatt tatctaattt
     1561 acacgaaatt gcttctggtg gcgctcccct ctctaaggaa gtcggggaag cggttgccaa
     1621 gaggttccat ctgccaggta agtgcactga gcagaaggga agaagctccg ggggctcttt
     1681 gtagggtgga tccagtgcat caaggcgatc acatcagtga aaaaaagcca gacaggcggt
     1741 taaaccaacg cagattaaac agcaggatgc aaaaattcgc aggtggtcag atggcggccg
     1801 cttctagact cgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     1861 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     1921 ttctcgagtt aagggcgaat tcccgataag gatcttccta gagcatggct acgtagataa
     1981 gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc
     2041 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     2101 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc cttaattaac ctaattcact
     2161 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     2221 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     2281 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     2341 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2401 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2461 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2521 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2581 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2641 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     2701 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     2761 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2821 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2881 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     2941 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3001 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3061 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3121 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     3181 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3241 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3301 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3361 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3421 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     3481 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3541 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     3601 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     3661 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     3721 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     3781 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     3841 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3901 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3961 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4021 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4081 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4141 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4201 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4261 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4321 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4381 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4441 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4501 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4561 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4621 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     4681 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     4741 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     4801 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     4861 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     4921 gaccatgatt acgccagatt taattaaggc cttaattagg ctgcgcgctc gctcgctcac
     4981 tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag
     5041 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct tgtagttaat
     5101 gattaacccg ccatgctact tatctacgta gccatgctct aggaagatcg gaattcgccc
     5161 ttaagggcgc gccgttt
//
