LOCUS       sequence-426613-        4776 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 4776)
  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 4776)
  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-BDLacZ-SAS620-3'Luciferase (split
            CAG...
FEATURES             Location/Qualifiers
     source          1..4776
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        101..480
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        481..684
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     635..655
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1522..1603
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1547..1566
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   1691..1820
                     /label=AAV2 ITR
     primer_bind     complement(1839..1856)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1839..1855)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1848..1870)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1997..2452
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2084..2103)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2294..2315
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2478..2582
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2583..2651,2652..3443)
                     /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(2801..2820)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3614..4202
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4103..4122
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4490..4511
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4526..4531,4532..4549,4550..4556)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4564..4580
                     /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     4569..4591
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4588..4604
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4588..4604
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   4642..4771
                     /label=AAV2 ITR
ORIGIN
        1 ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa gatcggaatt
       61 cgcccttaag ggcgcgccgt ttaaatagct agcacgcgtt gacattgatt attgactagt
      121 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      181 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      241 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      301 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      361 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      421 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      481 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      541 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      601 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      661 tgggaggtct atataagcag agctcgttta gtgaaccgtc agaggtacca ccggtcgact
      721 agaaggtgcg gccgccatct gaccacctgc gaatttttgc atcctgctgt ttaatctgcg
      781 ttggtttaac cgcctgtctg gctttttttc actgatgtga tcgccttgat gcactggatc
      841 ccaacgagtc ttttgtcatc tacaggtatc aggcaaggat atgggctcac tgagactaca
      901 tcagctattc tgattacacc cgagggggat gataaaccgg gcgcggtcgg taaagttgtt
      961 ccattttttg aagcgaaggt tgtggatctg gataccggga aaacgctggg cgttaatcaa
     1021 agaggcgaac tgtgtgtgag aggtcctatg attatgtccg gttatgtaaa caatccggaa
     1081 gcgaccaacg ccttgattga caaggatgga tggctacatt ctggagacat agcttactgg
     1141 gacgaagacg aacacttctt catcgttgac cgcctgaagt ctctgattaa gtacaaaggc
     1201 tatcaggtgg ctcccgctga attggaatcc atcttgctcc aacaccccaa catcttcgac
     1261 gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac ttcccgccgc cgttgttgtt
     1321 ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg attacgtcgc cagtcaagta
     1381 acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg acgaagtacc gaaaggtctt
     1441 accggaaaac tcgacgcaag aaaaatcaga gagatcctca taaaggccaa gaagggcgga
     1501 aagatcgccg tgtaatctag agcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     1561 tattgcttta tttgtaacca ttataagctg caataaacaa gttctcgagt taagggcgaa
     1621 ttcccgataa ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc
     1681 attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     1741 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     1801 gtgagcgagc gagcgcgcag ccttaattaa cctaattcac tggccgtcgt tttacaacgt
     1861 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     1921 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     1981 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2041 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     2101 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2161 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     2221 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2281 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     2341 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     2401 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     2461 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2521 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2581 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2641 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2701 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2761 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2821 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2881 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2941 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3001 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3061 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3121 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3181 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     3241 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3301 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3361 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3421 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     3481 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3541 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3601 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3661 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3721 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     3781 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3841 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3901 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3961 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4021 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4081 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4141 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4201 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4261 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     4321 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     4381 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     4441 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     4501 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     4561 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccagat
     4621 ttaattaagg ccttaattag gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag
     4681 cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag
     4741 ggagtggcca actccatcac taggggttcc ttgtag
//
