LOCUS       sequence-254649-        5241 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of AcrIIC3Nme fused to mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5241)
  AUTHORS   Lee J, Mou H, Ibraheim R, Liang SQ, Liu P, Xue W, Sontheimer EJ
  TITLE     Tissue-restricted Genome Editing in vivo Specified by 
            MicroRNA-repressible Anti-CRISPR Proteins.
  JOURNAL   RNA. 2019 Aug 22. pii: rna.071704.119. doi: 10.1261/rna.071704.119.
  PUBMED    31439808
REFERENCE   2  (bases 1 to 5241)
  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: "RNA. 2019 
            Aug 22. pii: rna.071704.119. doi: 10.1261/rna.071704.119."
COMMENT     Sequence Label: p1146-pCSDest-AcrIIC3Nme-mCherry
FEATURES             Location/Qualifiers
     source          1..5241
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    23..157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(47..66)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     101..120
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(270..290)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(270..288)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(309..325)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(309..325)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(322..344)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    333..349
                     /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)."
     promoter        complement(join(357..363,364..381,382..387))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    402..423
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(540..557)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(711..1299)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(791..810)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1470..2261,2262..2330))
                     /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     2093..2112
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2331..2435)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2461..2916)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2598..2619)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2810..2829
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2987..3971
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        4006..4024
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4006..4023
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4416..4439
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4446..4466
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4479..5189
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYYAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4624..4642)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4892..4911
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        complement(join(5241,1..18))
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ctatagtgag tcgtattacg tagatccaga catgataaga tacattgatg agtttggaca
       61 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
      121 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
      181 tatgtttcag gttcaggggg aggtgtggga ggttttttaa ttcgcggccg cggcgccaat
      241 gcattgggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
      301 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
      361 atacgagccg ggagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
      421 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
      481 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
      541 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
      601 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
      661 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
      721 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
      781 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      841 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      901 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      961 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1021 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1081 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1141 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1201 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1261 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1321 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     1381 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     1441 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     1501 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     1561 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     1621 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     1681 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     1741 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     1801 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     1861 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     1921 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1981 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     2041 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     2101 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     2161 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     2221 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     2281 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     2341 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     2401 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     2461 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     2521 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     2581 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     2641 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     2701 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     2761 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     2821 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     2881 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
     2941 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagtcg accgccaatt
     3001 caatatggcg tatatggact catgccaatt caatatggtg gatctggacc tgtgccaatt
     3061 caatatggcg tatatggact cgtgccaatt caatatggtg gatctggacc ccagccaatt
     3121 caatatggcg gacttggcac catgccaatt caatatggcg gacctggcac tgtgccaact
     3181 ggggaggggt ctacttggca cggtgccaag tttgaggagg ggtcttggcc ctgtgccaag
     3241 tccgccatat tgaattggca tggtgccaat aatggcggcc atattggcta tatgccagga
     3301 tcaatatata ggcaatatcc aatatggccc tatgccaata tggctattgg ccaggttcaa
     3361 tactatgtat tggccctatg ccatatagta ttccatatat gggttttcct attgacgtag
     3421 atagcccctc ccaatgggcg gtcccatata ccatatatgg ggcttcctaa taccgcccat
     3481 agccactccc ccattgacgt caatggtctc tatatatggt ctttcctatt gacgtcatat
     3541 gggcggtcct attgacgtat atggcgcctc ccccattgac gtcaattacg gtaaatggcc
     3601 cgcctggctc aatgcccatt gacgtcaata ggaccaccca ccattgacgt caatgggatg
     3661 gctcattgcc cattcatatc cgttctcacg ccccctattg acgtcaatga cggtaaatgg
     3721 cccacttggc agtacatcaa tatctattaa tagtaacttg gcaagtacat tactattgga
     3781 agtacgccag ggtacattgg cagtactccc attgacgtca atggcggtaa atggcccgcg
     3841 atggctgcca agtacatccc cattgacgtc aatggggagg ggcaatgacg caaatgggcg
     3901 ttccattgac gtaaatgggc ggtaggcgtg cctaatggga ggtctatata agcaatgctc
     3961 gtttagggaa ccgccattct gcctggggac gtcggagcaa gcttgattta ggtgacacta
     4021 tagacggaat tgtacccgcg gccgatccac cggtcgccac catgttcaaa cgggccatta
     4081 ttttcaccag cttcaacggc ttcgagaagg tcagtcgcac ggaaaaacgg cgccttgcca
     4141 agataattaa cgccagagtt agtataatcg acgagtatct tcgcgccaaa gacaccaacg
     4201 cctccttgga cggacagtat agagcattcc ttttcaatga tgagtcacca gcgatgaccg
     4261 aattcctcgc taagctcaag gcgttcgcag aaagctgtac tggtataagc attgatgcgt
     4321 gggagattga agagagcgag tatgttcgat tgcccgtcga acggagagat ttccttgcgg
     4381 cggccaatgg caaagaaatt ttcaaaattg gcggggatta taaggatgat gatgataagg
     4441 gctcccctaa gaagaagaga aaggtgggat ccggaagtat ggtgagcaag ggcgaggagg
     4501 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
     4561 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     4621 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
     4681 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
     4741 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     4801 gcgtggtgac cgtgacccag gactcctcac tgcaggacgg cgagttcatc tacaaggtga
     4861 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
     4921 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
     4981 agaggctgaa gctgaaggac ggcggccact actacgctga ggtcaagacc acctacaagg
     5041 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
     5101 cccacaacga ggactacacc atcgtggaac agtacgaacg cgctgagggc cgccactcca
     5161 ccggcggcat ggacgagctg tacaagtgat aaagcggcca tcaagcttat cgataccgtc
     5221 gactagagct cgctgatcag c
//
