LOCUS       sequence-214722-        7296 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 7296)
  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 7296)
  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..7296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2069..2090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2281..2300
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2514..2536
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2528..2545
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2529..2545
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   2564..2693
                     /label=AAV2 ITR
     polyA_signal    2855..2976
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2892..2911)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2946..2965
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    3054..3642
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3160..3171
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3569..3589
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    3714..3747
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3818..3851
                     /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             5277..5984
                     /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(5422..5440)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5690..5709
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(6042..6075)
                     /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(6138..6171)
                     /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(6182..6201)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          6184..6316
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     repeat_region   6909..7038
                     /label=AAV2 ITR
     primer_bind     complement(7076..7092)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7076..7092)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7089..7111)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7100..7116
                     /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(7124..7130,7131..7148,7149..7154))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7169..7190
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     2521 acgacgttgt aaaacgacgg ccagtgaatt aggttaatta aggctgcgcg ctcgctcgct
     2581 cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
     2641 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt ccttgtagtt
     2701 aatgattaac ccgccatgct acttatctac gtagccatgc tctaggaaga tccttatcga
     2761 ttttaccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacatc tccccctgaa
     2821 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2881 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2941 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgctcga agcggccgcc
     3001 cgggtcgact ctagatgcat gctcgagcgg ccgcccgggt cgacggtatc gatgcgggga
     3061 ggcggcccaa agggagatcc gactcgtctg agggcgaagg cgaagacgcg gaagaggccg
     3121 cagagccggc agcaggccgc gggaaggaag gtccgctgga ttgagggccg aagggacgta
     3181 gcagaaggac gtcccgcgca gaatccaggt ggcaacacag gcgagcagcc aaggaaagga
     3241 cgatgatttc cccgacaaca ccacggaatt gtcagtgccc aacagccgag cccctgtcca
     3301 gcagcgggca aggcaggcgg cgatgagttc cgccgtggca atagggaggg ggaaagcgaa
     3361 agtcccggaa aggagctgac aggtggtggc aatgccccaa ccagtggggg ttgcgtcagc
     3421 aaacacagtg cacaccacgc cacgttgcct gacaacgggc cacaactcct cataaagaga
     3481 cagcaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag caatagcatg
     3541 atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat agttaagaat
     3601 accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttatcgataa gcttctcgag
     3661 tggatcatct gcagaattcg cccttttcag tgtttcgaac ctcttcgagg gacataactt
     3721 cgtataatgt atgctatacg aagttatggg tcgatggtga gatctggact agagggtcga
     3781 tggtgatgct tggatcaaaa taggaagaat cccaatgata acttcgtata aagtatccta
     3841 tacgaagtta tactagtcat atgggatccg aattctgaac tagtgcaggt atggatatca
     3901 tctcggtggc attgaaacgt cattctacca aagccttcga tgcgtcgaaa aagcttacgc
     3961 cagagcaagc tgaacagatc aagacgttgc ttcagtattc acccagttca actaattccc
     4021 aaccctggca cttcatcgtg gcgtccacgg aggaagggaa agcgcgagta gcgaaatcag
     4081 cagccggcaa ctatgtcttt aacgaacgca aaatgctcga tgcaagccac gtggtggtct
     4141 tttgtgccaa aaccgcaatg gacgatgtat ggcttaagct ggtagtggac caagaggatg
     4201 cggatggaag gttcgctaca ccggaggcga aagccgcgaa tgataaggga agaaagttct
     4261 gggcagacat gcatcgaaaa gacctccatg atgacgcgga gtggatggcc aagcaggttt
     4321 accttaatgt aggtaacttc ctgttggggg tcgccgcgtt ggggctggac gctgtgccta
     4381 ttgaaggttt tgacgccgcc attttggatg cagagtttgg actcaaagaa aaaggctaca
     4441 ctagcctcgt ggtggttccg gtcggtcacc actcggtgga ggactttaac gcgaccctcc
     4501 ccaagtcgcg gctgccgcag aatatcacac tcacggaagt ggctggttct agtgcaggtc
     4561 aaggtgcaca ggctggtcaa ggagcacagg ctggttctag tgcaggtatg gatatcatct
     4621 cggtggcatt gaaacgtcat tctaccaaag ccttcgatgc gtcgaaaaag cttacgccag
     4681 agcaagctga acagatcaag acgttgcttc agtattcacc cagttcaact aattcccaac
     4741 cctggcactt catcgtggcg tccacggagg aagggaaagc gcgagtagcg aaatcagcag
     4801 ccggcaacta tgtctttaac gaacgcaaaa tgctcgatgc aagccacgtg gtggtctttt
     4861 gtgccaaaac cgcaatggac gatgtatggc ttaagctggt agtggaccaa gaggatgcgg
     4921 atggaaggtt cgctacaccg gaggcgaaag ccgcgaatga taagggaaga aagttctggg
     4981 cagacatgca tcgaaaagac ctccatgatg acgcggagtg gatggccaag caggtttacc
     5041 ttaatgtagg taacttcctg ttgggggtcg ccgcgttggg gctggacgct gtgcctattg
     5101 aaggttttga cgccgccatt ttggatgcag agtttggact caaagaaaaa ggctacacta
     5161 gcctcgtggt ggttccggtc ggtcaccact cggtggagga ctttaacgcg accctcccca
     5221 agtcgcggct gccgcagaat atcacactca cggaagtggc tggttctagt gcaggtatgg
     5281 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     5341 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     5401 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     5461 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     5521 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     5581 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     5641 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     5701 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     5761 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     5821 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     5881 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     5941 ccgagggccg ccactccacc ggcggcatgg acgagctgta caaggctggt tctagtctcg
     6001 agtaagatat caccggtacg cgttgatcag ctagcactag tataacttcg tatagcatac
     6061 attatacgaa gttatgtgtc gatggtgatg cttggcaatt cgggtcgatg gtgaagcatt
     6121 ggtcttccta ttttgatata acttcgtata ggatacttta tacgaagtta tggggatcat
     6181 cacctgtgga gagaaaggca aagtggatgt cagtaagacc aataggtgcc tatcagaaac
     6241 gcaagagtct tctctgtctc gacaagccca gtttctattg gtctccttaa acctgtcttg
     6301 taaccttgat acttacctga tccaacggct gcgacttggg gcagggggtc ctaggggtgg
     6361 tctggccagg agccgcgggg gcggactgcg cggtgcccag gagcacagct gcgctctcag
     6421 gcacgacacg actcctccgc tgcccaccgc agactgaggc agcgctgagt cgccggcgcc
     6481 gcagcgcaga tggtcgcgcc cgtgcccccc tatctcgcgc ctcgcgtggt gcggtccggc
     6541 tgggccggcg gcggcgcgga cgcgaccaag gtggccggga aggggagttt gcgggggacc
     6601 ggcgagtgac gtcagcgcgc cttcagtgct gaggcggcgg tggcgcgcgc cgccaggcgg
     6661 gggcgaaggc actgtccgcg gtgctgaagc tggcagtgcg cacgcgcctc gccgcatcct
     6721 gtttcccctc cccctctctg ataggggatg cgcaatttgg ggaatggggg ttgggtgctt
     6781 gtccagtggg tcggggtcgg tcgtcaattt aaatactgaa aagggcgaat tccagcacac
     6841 tggagctcgg atcttcctag agcatggcta cgtagataag tagcatggcg ggttaatcat
     6901 taactacaag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     6961 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     7021 gagcgagcga gcgcgcagcc taattaaggc cttaattaaa tctggcgtaa tcatggtcat
     7081 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     7141 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     7201 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     7261 aacgcgcggg gagaggcggt ttgcgtattg ggcgct
//
