LOCUS       sequence-426610-        4691 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 4691)
  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 4691)
  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-BD lacZ_3' Cer-bGHpA
FEATURES             Location/Qualifiers
     source          1..4691
                     /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(1346..1365)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1505..1756
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH
                     YLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     1693..1714
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1766..1990
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1999..2128
                     /label=AAV2 ITR
     primer_bind     complement(2147..2164)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2147..2163)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2156..2178)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2305..2760
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2392..2411)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2602..2623
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2786..2890
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2891..2959,2960..3751)
                     /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(3109..3128)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3922..4510
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4411..4430
                     /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 taagcttgga tcccatctga ccacctgcga
     1081 atttttgcat cctgctgttt aatctgcgtt ggtttaaccg cctgtctggc tttttttcac
     1141 tgatgtgatc gccttgatgc actggatccc aacgagtctt ttgtcatcta caggtaagct
     1201 gcccgtgccc tggcccaccc tcgtgaccac cctgacctgg ggcgtgcagt gcttcgcccg
     1261 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1321 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     1381 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1441 cggcaacatc ctggggcaca agctggagta caacgccatc agcgacaacg tctatatcac
     1501 cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga
     1561 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1621 gctgctgccc gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga
     1681 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1741 ggacgagctg tacaagtaag tcgacctgtg ccttctagtt gccagccatc tgttgtttgc
     1801 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     1861 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     1921 gggcaggaca gcaaggtgga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     1981 ggctctatgg gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     2041 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     2101 cggcctcagt gagcgagcga gcgcgcagcc ttaattaacc taattcactg gccgtcgttt
     2161 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     2221 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     2281 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2341 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2401 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2461 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2521 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2581 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2641 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2701 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     2761 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2821 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2881 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2941 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3001 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3061 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3121 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3181 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3241 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3301 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3361 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3421 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3481 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3541 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3601 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3661 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3721 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3781 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3841 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3901 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3961 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4021 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4081 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4141 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4201 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4261 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4321 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4381 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4441 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4501 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4561 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4621 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4681 ggaagcggaa g
//
