LOCUS       sequence-426609-        4305 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Split fluorophore 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 4305)
  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 4305)
  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: pAAV2.1-CMV-5'Cer-BDlacZ-bGHpA
FEATURES             Location/Qualifiers
     source          1..4305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        385..764
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        765..968
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     919..939
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1013..1032
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1013..1031
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1103..1124)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    1380..1604
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1613..1742
                     /label=AAV2 ITR
     primer_bind     complement(1761..1778)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1761..1777)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1770..1792)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1919..2374
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2006..2025)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2216..2237
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2400..2504
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2505..2573,2574..3365)
                     /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(2723..2742)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3536..4124
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4025..4044
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccagattta
      241 attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      301 gggcggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
      361 gttcctgcgg cctctagaaa gcttgacatt gattattgac tagttattaa tagtaatcaa
      421 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      481 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      541 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      601 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      661 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      721 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      781 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      841 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      901 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      961 gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga
     1021 ctcactatag ggagacccaa gctggctagt taagcttatg gtgagcaagg gcgaggagct
     1081 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1141 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     1201 ctgcaccaca ggtaagggca ctgagcagaa gggaagaagc tccgggggct ctttgtaggg
     1261 tggatccagt gcatcaaggc gatcacatca gtgaaaaaaa gccagacagg cggttaaacc
     1321 aacgcagatt aaacagcagg atgcaaaaat tcgcaggtgg tcagatggga tccgtcgacc
     1381 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     1441 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     1501 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     1561 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atgggcggcc gcaggaaccc
     1621 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     1681 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     1741 agccttaatt aacctaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     1801 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     1861 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     1921 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1981 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2041 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2101 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2161 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2221 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2281 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2341 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     2401 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2461 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2521 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2581 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2641 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2701 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2761 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2821 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2881 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2941 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3001 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3061 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3121 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3181 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3241 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3301 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3361 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3421 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3481 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3541 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3601 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3661 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3721 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3781 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3841 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3901 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3961 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4021 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4081 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4141 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4201 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4261 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaag
//
