LOCUS       sequence-254650-        5154 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of AcrIIA4 fused to mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5154)
  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 5154)
  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: p1148-pCSDest-AcrIIA4-mCherry
FEATURES             Location/Qualifiers
     source          1..5154
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4..22
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4..21
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             327..350
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             357..377
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             390..1100
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(535..553)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     803..822
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        complement(1152..1170)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1175..1309
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1199..1218)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1253..1272
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1422..1442)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1422..1440)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1461..1477)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1461..1477)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1474..1496)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1485..1501
                     /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(1509..1515,1516..1533,1534..1539))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1554..1575
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1692..1709)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1863..2451)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1943..1962)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2622..3413,3414..3482))
                     /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     3245..3264
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3483..3587)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3613..4068)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3750..3771)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3962..3981
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4139..5123
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 ttgatttagg tgacactata gacggaattg tacccgcggc cgatccaccg gtcgccacca
       61 tgaacatcaa tgatctgatt agagaaataa agaataaaga ctatactgtt aaattgtctg
      121 gaactgacag taatagcata acccaactca tcatcagggt taataatgat ggtaacgaat
      181 atgttataag tgagtccgag aacgaatcta tcgtcgagaa gttcatcagt gccttcaaaa
      241 acggatggaa ccaagagtac gaggatgagg aggaatttta caatgatatg caaacaatca
      301 ctctgaagag cgagcttaac ggcggggatt ataaggatga tgatgataag ggctccccta
      361 agaagaagag aaaggtggga tccggaagta tggtgagcaa gggcgaggag gataacatgg
      421 ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg
      481 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
      541 tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca
      601 tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt
      661 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga
      721 ccgtgaccca ggactcctca ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg
      781 gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct
      841 cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga
      901 agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc
      961 ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg
     1021 aggactacac catcgtggaa cagtacgaac gcgctgaggg ccgccactcc accggcggca
     1081 tggacgagct gtacaagtga taaagcggcc atcaagctta tcgataccgt cgactagagc
     1141 tcgctgatca gcctatagtg agtcgtatta cgtagatcca gacatgataa gatacattga
     1201 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     1261 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     1321 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aattcgcggc
     1381 cgcggcgcca atgcattggg cccggtaccc agcttttgtt ccctttagtg agggttaatt
     1441 gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     1501 attccacaca acatacgagc cgggagcata aagtgtaaag cctggggtgc ctaatgagtg
     1561 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     1621 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     1681 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     1741 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     1801 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1861 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     1921 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     1981 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2041 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2101 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2161 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2221 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2281 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2341 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2401 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2461 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2521 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2581 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     2641 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     2701 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     2761 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     2821 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     2881 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     2941 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3001 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3061 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3121 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3181 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3241 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3301 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3361 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3421 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3481 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     3541 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     3601 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3661 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3721 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3781 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3841 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3901 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3961 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     4021 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat
     4081 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt
     4141 cgaccgccaa ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg tggatctgga
     4201 cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga
     4261 ccccagccaa ttcaatatgg cggacttggc accatgccaa ttcaatatgg cggacctggc
     4321 actgtgccaa ctggggaggg gtctacttgg cacggtgcca agtttgagga ggggtcttgg
     4381 ccctgtgcca agtccgccat attgaattgg catggtgcca ataatggcgg ccatattggc
     4441 tatatgccag gatcaatata taggcaatat ccaatatggc cctatgccaa tatggctatt
     4501 ggccaggttc aatactatgt attggcccta tgccatatag tattccatat atgggttttc
     4561 ctattgacgt agatagcccc tcccaatggg cggtcccata taccatatat ggggcttcct
     4621 aataccgccc atagccactc ccccattgac gtcaatggtc tctatatatg gtctttccta
     4681 ttgacgtcat atgggcggtc ctattgacgt atatggcgcc tcccccattg acgtcaatta
     4741 cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa taggaccacc caccattgac
     4801 gtcaatggga tggctcattg cccattcata tccgttctca cgccccctat tgacgtcaat
     4861 gacggtaaat ggcccacttg gcagtacatc aatatctatt aatagtaact tggcaagtac
     4921 attactattg gaagtacgcc agggtacatt ggcagtactc ccattgacgt caatggcggt
     4981 aaatggcccg cgatggctgc caagtacatc cccattgacg tcaatgggga ggggcaatga
     5041 cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata
     5101 taagcaatgc tcgtttaggg aaccgccatt ctgcctgggg acgtcggagc aagc
//
