LOCUS       sequence-218207-        7169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated conditional expression of bacterial tdNfsB-mCherry 
            (eNTR) under the CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7169)
  AUTHORS   Gruber TD, Krishnamurthy C, Grimm JB, Tadross MR, Wysocki LM, 
            Gartner ZJ, Lavis LD
  TITLE     Cell-Specific Chemical Delivery Using a Selective 
            Nitroreductase-Nitroaryl Pair.
  JOURNAL   ACS Chem Biol. 2018 Aug 29. doi: 10.1021/acschembio.8b00524.
  PUBMED    30111097
REFERENCE   2  (bases 1 to 7169)
  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: "ACS Chem 
            Biol. 2018 Aug 29. doi: 10.1021/acschembio.8b00524."
COMMENT     Sequence Label: aav-CAG-FLEX-tdNfsB(F124W)-mCherry (eNTR)
FEATURES             Location/Qualifiers
     source          1..7169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1356..2063
                     /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(1501..1519)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1769..1788
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(2117..2150)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(2213..2246)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(2284..2303)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(2365..2387)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(2666..2943)
                     /label=chicken beta-actin promoter
     repeat_region   3254..3383
                     /label=AAV2 ITR
     rep_origin      3455..3910
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3542..3561)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3752..3773
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3936..4040
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4041..4109,4110..4901)
                     /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(4259..4278)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5072..5660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5561..5580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5897..6026
                     /label=AAV2 ITR
     polyA_signal    6188..6309
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6225..6244)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6279..6298
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    6352..6940
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6458..6469
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     6867..6887
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    6974..7007
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    7078..7111
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
ORIGIN
        1 attctaccaa agccttcgat gcgtcgaaaa agcttacgcc agagcaagct gaacagatca
       61 agacgttgct tcagtattca cccagttcaa ctaattccca accctggcac ttcatcgtgg
      121 cgtccacgga ggaagggaaa gcgcgagtag cgaaatcagc agccggcaac tatgtcttta
      181 acgaacgcaa aatgctcgat gcaagccacg tggtggtctt ttgtgccaaa accgcaatgg
      241 acgatgtatg gcttaagctg gtagtggacc aagaggatgc ggatggaagg ttcgctacac
      301 cggaggcgaa agccgcgaat gataagggaa gaaagttctg ggcagacatg catcgaaaag
      361 acctccatga tgacgcggag tggatggcca agcaggttta ccttaatgta ggtaacttcc
      421 tgttgggggt cgccgcgttg gggctggacg ctgtgcctat tgaaggtttt gacgccgcca
      481 ttttggatgc agagtttgga ctcaaagaaa aaggctacac tagcctcgtg gtggttccgg
      541 tcggtcacca ctcggtggag gactttaacg cgaccctccc caagtcgcgg ctgccgcaga
      601 atatcacact cacggaagtg gctggttcta gtgcaggtca aggtgcacag gctggtcaag
      661 gagcacaggc tggttctagt gcaggtatgg atatcatctc ggtggcattg aaacgtcatt
      721 ctaccaaagc cttcgatgcg tcgaaaaagc ttacgccaga gcaagctgaa cagatcaaga
      781 cgttgcttca gtattcaccc agttcaacta attcccaacc ctggcacttc atcgtggcgt
      841 ccacggagga agggaaagcg cgagtagcga aatcagcagc cggcaactat gtctttaacg
      901 aacgcaaaat gctcgatgca agccacgtgg tggtcttttg tgccaaaacc gcaatggacg
      961 atgtatggct taagctggta gtggaccaag aggatgcgga tggaaggttc gctacaccgg
     1021 aggcgaaagc cgcgaatgat aagggaagaa agttctgggc agacatgcat cgaaaagacc
     1081 tccatgatga cgcggagtgg atggccaagc aggtttacct taatgtaggt aacttcctgt
     1141 tgggggtcgc cgcgttgggg ctggacgctg tgcctattga aggttttgac gccgccattt
     1201 tggatgcaga gtttggactc aaagaaaaag gctacactag cctcgtggtg gttccggtcg
     1261 gtcaccactc ggtggaggac tttaacgcga ccctccccaa gtcgcggctg ccgcagaata
     1321 tcacactcac ggaagtggct ggttctagtg caggtatggt gagcaagggc gaggaggata
     1381 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     1441 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     1501 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     1561 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     1621 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     1681 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     1741 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     1801 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
     1861 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
     1921 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
     1981 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     2041 gcggcatgga cgagctgtac aaggctggtt ctagtctcga gtaagatatc accggtggcg
     2101 cgccaagctt gctagtataa cttcgtatag catacattat acgaagttat gggtcgatgg
     2161 tgatgcttgg caattcgggt cgatggtgaa gcattggtct tcctattttg atataacttc
     2221 gtataggata ctttatacga agttatgggg atgatccaat tctttgccaa aatgatgaga
     2281 cagcacaata accagcacgt tgcccaggag ctgtaggaaa aagaagaagg catgaacatg
     2341 gttagcagag gctctagagc cgccggtcac acgccagaag ccgaaccccg ccctgccccg
     2401 tcccccccga aggcagccgt ccccccgcgg acagccccga gccgctcccc ccgcacaaag
     2461 ggccctcccg gagcccctca aggctttcac gcagccacag aaaagaaaca agccgtcatt
     2521 aaaccaagcg ctaattacag cccggaggag aagggccgtc ccgcccgctc acctgtggga
     2581 gtaacgcggt cagtcagagc cggggcgggc ggcgcgaggc ggcggcggag cggggcacgg
     2641 ggcgaaggca gcgcgcagcg actcccgccc gccgcgcgct tcgcttttta tagggccgcc
     2701 gccgccgccg cctcgccata aaaggaaact ttcggagcgc gccgctctga ttggctgccg
     2761 ccgcacctct ccgcctcgcc ccgccccgcc cctcgccccg ccccgccccg cctggcgcgc
     2821 gccccccccc cccccccgcc cccatcgctg cacaaaataa ttaaaaaata aataaataca
     2881 aaattggggg tggggagggg ggggagatgg ggagagtgaa gcagaacgtg gggctcacct
     2941 cgaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata
     3001 aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg
     3061 gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact
     3121 ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta
     3181 ttgacgtcaa tggtactctt cctagagcat ggctacgtag ataagtagca tggcgggtta
     3241 atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3301 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     3361 tcagtgagcg agcgagcgcg cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     3421 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     3481 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3541 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3601 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3661 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3721 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3781 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3841 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     3901 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3961 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4021 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4081 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4141 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4201 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4261 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4321 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4381 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4441 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4501 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4561 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     4621 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4681 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4741 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4801 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4861 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     4921 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4981 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5041 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5101 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5161 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5221 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5281 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5341 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5401 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5461 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5521 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5581 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5641 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5701 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5761 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     5821 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     5881 cgattcatta atgcagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg
     5941 gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt
     6001 ggccaactcc atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc
     6061 tacgtagcca tgctctagga agatccttat cgattttacc acatttgtag aggttttact
     6121 tgctttaaaa aacctcccac atctccccct gaacctgaaa cataaaatga atgcaattgt
     6181 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     6241 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     6301 tgtatcttat catgtctgct cgaagcggcc gcccgggtcg acggtatcga tgcggggagg
     6361 cggcccaaag ggagatccga ctcgtctgag ggcgaaggcg aagacgcgga agaggccgca
     6421 gagccggcag caggccgcgg gaaggaaggt ccgctggatt gagggccgaa gggacgtagc
     6481 agaaggacgt cccgcgcaga atccaggtgg caacacaggc gagcagccaa ggaaaggacg
     6541 atgatttccc cgacaacacc acggaattgt cagtgcccaa cagccgagcc cctgtccagc
     6601 agcgggcaag gcaggcggcg atgagttccg ccgtggcaat agggaggggg aaagcgaaag
     6661 tcccggaaag gagctgacag gtggtggcaa tgccccaacc agtgggggtt gcgtcagcaa
     6721 acacagtgca caccacgcca cgttgcctga caacgggcca caactcctca taaagagaca
     6781 gcaaccagga tttatacaag gaggagaaaa tgaaagccat acgggaagca atagcatgat
     6841 acaaaggcat taaagcagcg tatccacata gcgtaaaagg agcaacatag ttaagaatac
     6901 cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt atcgataagc taattcgaac
     6961 ctcttcgagg gacataactt cgtataatgt atgctatacg aagttatggg tcgatggtga
     7021 gatctggact agagggtcga tggtgatgct tggatcaaaa taggaagaat cccaatgata
     7081 acttcgtata aagtatccta tacgaagtta tggatccgaa ttctgaacta gtgcaggtat
     7141 ggatatcatc tcggtggcat tgaaacgtc
//
