LOCUS       sequence-214727-        7172 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 7172)
  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 7172)
  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..7172
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   56..185
                     /label=AAV2 ITR
     polyA_signal    347..468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(384..403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     438..457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    511..1099
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             617..628
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1026..1046
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    1133..1166
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1237..1270
                     /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)."
     CDS             2684..3391
                     /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(2829..2847)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3097..3116
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(3445..3478)
                     /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(3541..3574)
                     /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(3612..3631)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3693..3715)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3995..4272)
                     /label=chicken beta-actin promoter
     repeat_region   4585..4714
                     /label=AAV2 ITR
     rep_origin      4786..5241
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4873..4892)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5083..5104
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5267..5371
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5372..5440,5441..6232)
                     /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(5590..5609)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6403..6991
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6892..6911
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      121 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
      241 gatccttatc gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca
      301 tctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
      361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
      421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc
      481 gaagcggccg cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
      541 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
      601 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
      661 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
      721 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
      781 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
      841 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
      901 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
      961 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     1021 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     1081 tgtaatccag aggttgatta tcgataagct aattcgaacc tcttcgaggg acataacttc
     1141 gtataatgta tgctatacga agttatgggt cgatggtgag atctggacta gagggtcgat
     1201 ggtgatgctt ggatcaaaat aggaagaatc ccaatgataa cttcgtataa agtatcctat
     1261 acgaagttat ggatccgaat tctgaactag tgcaggtatg gatatcatct cggtggcatt
     1321 gaaacgtcat tctaccaaag ccttcgatgc gtcgaaaaag cttacgccag agcaagctga
     1381 acagatcaag acgttgcttc agtattcacc cagttcaact aattcccaac cctggcactt
     1441 catcgtggcg tccacggagg aagggaaagc gcgagtagcg aaatcagcag ccggcaacta
     1501 tgtctttaac gaacgcaaaa tgctcgatgc aagccacgtg gtggtctttt gtgccaaaac
     1561 cgcaatggac gatgtatggc ttaagctggt agtggaccaa gaggatgcgg atggaaggtt
     1621 cgctacaccg gaggcgaaag ccgcgaatga taagggaaga aagttctggg cagacatgca
     1681 tcgaaaagac ctccatgatg acgcggagtg gatggccaag caggtttacc ttaatgtagg
     1741 taacttcctg ttgggggtcg ccgcgttggg gctggacgct gtgcctattg aaggttttga
     1801 cgccgccatt ttggatgcag agtttggact caaagaaaaa ggctacacta gcctcgtggt
     1861 ggttccggtc ggtcaccact cggtggagga ctttaacgcg accctcccca agtcgcggct
     1921 gccgcagaat atcacactca cggaagtggc tggttctagt gcaggtcaag gtgcacaggc
     1981 tggtcaagga gcacaggctg gttctagtgc aggtatggat atcatctcgg tggcattgaa
     2041 acgtcattct accaaagcct tcgatgcgtc gaaaaagctt acgccagagc aagctgaaca
     2101 gatcaagacg ttgcttcagt attcacccag ttcaactaat tcccaaccct ggcacttcat
     2161 cgtggcgtcc acggaggaag ggaaagcgcg agtagcgaaa tcagcagccg gcaactatgt
     2221 ctttaacgaa cgcaaaatgc tcgatgcaag ccacgtggtg gtcttttgtg ccaaaaccgc
     2281 aatggacgat gtatggctta agctggtagt ggaccaagag gatgcggatg gaaggttcgc
     2341 tacaccggag gcgaaagccg cgaatgataa gggaagaaag ttctgggcag acatgcatcg
     2401 aaaagacctc catgatgacg cggagtggat ggccaagcag gtttacctta atgtaggtaa
     2461 cttcctgttg ggggtcgccg cgttggggct ggacgctgtg cctattgaag gttttgacgc
     2521 cgccattttg gatgcagagt ttggactcaa agaaaaaggc tacactagcc tcgtggtggt
     2581 tccggtcggt caccactcgg tggaggactt taacgcgacc ctccccaagt cgcggctgcc
     2641 gcagaatatc acactcacgg aagtggctgg ttctagtgca ggtatggtga gcaagggcga
     2701 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
     2761 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     2821 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     2881 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     2941 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     3001 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     3061 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     3121 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     3181 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     3241 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     3301 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     3361 ctccaccggc ggcatggacg agctgtacaa ggctggttct agtctcgagt aagatatcac
     3421 cggtggcgcg ccaagcttgc tagtataact tcgtatagca tacattatac gaagttatgg
     3481 gtcgatggtg atgcttggca attcgggtcg atggtgaagc attggtcttc ctattttgat
     3541 ataacttcgt ataggatact ttatacgaag ttatggggat gatccaattc tttgccaaaa
     3601 tgatgagaca gcacaataac cagcacgttg cccaggagct gtaggaaaaa gaagaaggca
     3661 tgaacatggt tagcagaggc tctagagccg ccggtcacac gccagaagcc gaaccccgcc
     3721 ctgccccgtc ccccccgaag gcagccgtcc ccccgcggac agccccgagc cgctcccccc
     3781 gcacaaaggg ccctcccgga gcccctcaag gctttcacgc agccacagaa aagaaacaag
     3841 ccgtcattaa accaagcgct aattacagcc cggaggagaa gggccgtccc gcccgctcac
     3901 ctgtgggagt aacgcggtca gtcagagccg gggcgggcgg cgcgaggcgg cggcggagcg
     3961 gggcacgggg cgaaggcagc gtcgcagcga ctcccgcccg ccgcgcgctt cgctttttat
     4021 agggccgccg ccgccgccgc ctcgccataa aaggaaactt tcggagcgcg ccgctctgat
     4081 tggctgccgc cgcacctctc cgcctcgccc cgccccgccc ctcgccccgc cccgccccgc
     4141 ctggcgcgcg cccccccccc ccccccgccc ccatcgctgc acaaaataat taaaaaataa
     4201 ataaatacaa aattgggggt ggggaggggg gggagatggg gagagtgaag cagaacgtgg
     4261 ggctcacctc gaccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     4321 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     4381 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     4441 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     4501 acgtcattat tgacgtcaat ggtacgatct tcctagagca tggctacgta gataagtagc
     4561 atggcgggtt aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc
     4621 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     4681 cccgggcggc ctcagtgagc gagcgagcgc gcagctggcg taatagcgaa gaggcccgca
     4741 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     4801 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4861 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4921 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4981 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     5041 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     5101 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     5161 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     5221 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5281 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5341 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5401 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5461 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5521 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5581 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5641 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5701 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5761 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5821 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5881 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5941 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6001 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6061 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6121 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6181 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6241 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6301 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6361 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6421 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6481 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6541 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6601 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6661 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6721 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6781 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6841 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6901 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6961 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7021 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     7081 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     7141 gcgcagcgag tcagtgagcg aggaagcgga ag
//
