LOCUS       sequence-218258-        6988 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated conditional expression of bacterial tdNfsB-mCherry 
            (eNTR) under the SYN promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6988)
  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 6988)
  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-SYN-FLEX-tdNfsB(F124W)-mCherry (eNTR)
FEATURES             Location/Qualifiers
     source          1..6988
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    275..396
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(312..331)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     366..385
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    474..1062
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             580..591
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     989..1009
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    1134..1167
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1238..1271
                     /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             2697..3404
                     /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(2842..2860)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3110..3129
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(3462..3495)
                     /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(3558..3591)
                     /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(3602..3621)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          3604..3736
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     repeat_region   4329..4458
                     /label=AAV2 ITR
     rep_origin      4530..4985
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4617..4636)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4827..4848
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5011..5115
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5116..5184,5185..5976)
                     /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(5334..5353)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6147..6735
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6636..6655
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6972..6988,1..113)
                     /label=AAV2 ITR
ORIGIN
        1 cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
       61 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt ccttgtagtt
      121 aatgattaac ccgccatgct acttatctac gtagccatgc tctaggaaga tccttatcga
      181 ttttaccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacatc tccccctgaa
      241 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
      301 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
      361 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgctcga agcggccgcc
      421 cgggtcgact ctagatgcat gctcgagcgg ccgcccgggt cgacggtatc gatgcgggga
      481 ggcggcccaa agggagatcc gactcgtctg agggcgaagg cgaagacgcg gaagaggccg
      541 cagagccggc agcaggccgc gggaaggaag gtccgctgga ttgagggccg aagggacgta
      601 gcagaaggac gtcccgcgca gaatccaggt ggcaacacag gcgagcagcc aaggaaagga
      661 cgatgatttc cccgacaaca ccacggaatt gtcagtgccc aacagccgag cccctgtcca
      721 gcagcgggca aggcaggcgg cgatgagttc cgccgtggca atagggaggg ggaaagcgaa
      781 agtcccggaa aggagctgac aggtggtggc aatgccccaa ccagtggggg ttgcgtcagc
      841 aaacacagtg cacaccacgc cacgttgcct gacaacgggc cacaactcct cataaagaga
      901 cagcaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag caatagcatg
      961 atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat agttaagaat
     1021 accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttatcgataa gcttctcgag
     1081 tggatcatct gcagaattcg cccttttcag tgtttcgaac ctcttcgagg gacataactt
     1141 cgtataatgt atgctatacg aagttatggg tcgatggtga gatctggact agagggtcga
     1201 tggtgatgct tggatcaaaa taggaagaat cccaatgata acttcgtata aagtatccta
     1261 tacgaagtta tactagtcat atgggatccg aattctgaac tagtgcaggt atggatatca
     1321 tctcggtggc attgaaacgt cattctacca aagccttcga tgcgtcgaaa aagcttacgc
     1381 cagagcaagc tgaacagatc aagacgttgc ttcagtattc acccagttca actaattccc
     1441 aaccctggca cttcatcgtg gcgtccacgg aggaagggaa agcgcgagta gcgaaatcag
     1501 cagccggcaa ctatgtcttt aacgaacgca aaatgctcga tgcaagccac gtggtggtct
     1561 tttgtgccaa aaccgcaatg gacgatgtat ggcttaagct ggtagtggac caagaggatg
     1621 cggatggaag gttcgctaca ccggaggcga aagccgcgaa tgataaggga agaaagttct
     1681 gggcagacat gcatcgaaaa gacctccatg atgacgcgga gtggatggcc aagcaggttt
     1741 accttaatgt aggtaacttc ctgttggggg tcgccgcgtt ggggctggac gctgtgccta
     1801 ttgaaggttt tgacgccgcc attttggatg cagagtttgg actcaaagaa aaaggctaca
     1861 ctagcctcgt ggtggttccg gtcggtcacc actcggtgga ggactttaac gcgaccctcc
     1921 ccaagtcgcg gctgccgcag aatatcacac tcacggaagt ggctggttct agtgcaggtc
     1981 aaggtgcaca ggctggtcaa ggagcacagg ctggttctag tgcaggtatg gatatcatct
     2041 cggtggcatt gaaacgtcat tctaccaaag ccttcgatgc gtcgaaaaag cttacgccag
     2101 agcaagctga acagatcaag acgttgcttc agtattcacc cagttcaact aattcccaac
     2161 cctggcactt catcgtggcg tccacggagg aagggaaagc gcgagtagcg aaatcagcag
     2221 ccggcaacta tgtctttaac gaacgcaaaa tgctcgatgc aagccacgtg gtggtctttt
     2281 gtgccaaaac cgcaatggac gatgtatggc ttaagctggt agtggaccaa gaggatgcgg
     2341 atggaaggtt cgctacaccg gaggcgaaag ccgcgaatga taagggaaga aagttctggg
     2401 cagacatgca tcgaaaagac ctccatgatg acgcggagtg gatggccaag caggtttacc
     2461 ttaatgtagg taacttcctg ttgggggtcg ccgcgttggg gctggacgct gtgcctattg
     2521 aaggttttga cgccgccatt ttggatgcag agtttggact caaagaaaaa ggctacacta
     2581 gcctcgtggt ggttccggtc ggtcaccact cggtggagga ctttaacgcg accctcccca
     2641 agtcgcggct gccgcagaat atcacactca cggaagtggc tggttctagt gcaggtatgg
     2701 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     2761 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     2821 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     2881 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     2941 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     3001 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     3061 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     3121 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     3181 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     3241 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     3301 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     3361 ccgagggccg ccactccacc ggcggcatgg acgagctgta caaggctggt tctagtctcg
     3421 agtaagatat caccggtacg cgttgatcag ctagcactag tataacttcg tatagcatac
     3481 attatacgaa gttatgtgtc gatggtgatg cttggcaatt cgggtcgatg gtgaagcatt
     3541 ggtcttccta ttttgatata acttcgtata ggatacttta tacgaagtta tggggatcat
     3601 tacctgtgga gagaaaggca aagtggatgt cagtaagacc aataggtgcc tatcagaaac
     3661 gcaagagtct tctctgtctc gacaagccca gtttctattg gtctccttaa acctgtcttg
     3721 taaccttgat acttacctga tccaacggct gcgacttggg gcagggggtc ctaggggtgg
     3781 tctggccagg agccgcgggg gcggactgcg cggtgcccag gagcacagct gcgctctcag
     3841 gcacgacacg actcctccgc tgcccaccgc agactgaggc agcgctgagt cgccggcgcc
     3901 gcagcgcaga tggtcgcgcc cgtgcccccc tatctcgcgc ctcgcgtggt gcggtccggc
     3961 tgggccggcg gcggcgcgga cgcgaccaag gtggccggga aggggagttt gcgggggacc
     4021 ggcgagtgac gtcagcgcgc cttcagtgct gaggcggcgg tggcgcgcgc cgccaggcgg
     4081 gggcgaaggc actgtccgcg gtgctgaagc tggcagtgcg cacgcgcctc gccgcatcct
     4141 gtttcccctc cccctctctg ataggggatg cgcaatttgg ggaatggggg ttgggtgctt
     4201 gtccagtggg tcggggtcgg tcgtcaattt aaatactgaa aagggcgaat tccagcacac
     4261 tggagctcgg atcttcctag agcatggcta cgtagataag tagcatggcg ggttaatcat
     4321 taactacaag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     4381 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     4441 gagcgagcga gcgcgcagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     4501 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     4561 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4621 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4681 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4741 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4801 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4861 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4921 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     4981 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5041 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5101 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5161 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5221 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5281 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5341 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5401 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5461 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5521 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5581 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5641 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5701 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5761 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5821 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5881 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5941 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6001 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6061 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6121 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6181 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6241 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6301 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6361 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6421 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6481 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6541 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6601 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6661 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6721 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6781 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6841 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     6901 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     6961 cattaatgca gctgcgcgct cgctcgct
//
