LOCUS       sequence-218203-        6836 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated 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 6836)
  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 6836)
  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-tdNfsB(F124W)-mCherry (eNTR)
FEATURES             Location/Qualifiers
     source          1..6836
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   56..185
                     /label=AAV2 ITR
     promoter        498..775
                     /label=chicken beta-actin promoter
     primer_bind     1054..1076
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1138..1157
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2600..3307
                     /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(2745..2763)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3013..3032
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3335..3923
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3388..3408)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3806..3817)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3966..4087
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3977..3996)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4031..4050
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4249..4378
                     /label=AAV2 ITR
     rep_origin      4450..4905
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4537..4556)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4747..4768
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4931..5035
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5036..5104,5105..5896)
                     /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(5254..5273)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6067..6655
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6556..6575
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgkcgggcga cctttggtcg
      121 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
      241 gatcgtacca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      301 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      361 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      421 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      481 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc
      541 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg
      601 ggcggggggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg
      661 ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt
      721 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt
      781 cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg
      841 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
      901 ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct
      961 tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggctgt ccgcgggggg
     1021 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1081 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1141 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attggatcct aggaattctg
     1201 aactagtgca ggtatggata tcatctcggt ggcattgaaa cgtcattcta ccaaagcctt
     1261 cgatgcgtcg aaaaagctta cgccagagca agctgaacag atcaagacgt tgcttcagta
     1321 ttcacccagt tcaactaatt cccaaccctg gcacttcatc gtggcgtcca cggaggaagg
     1381 gaaagcgcga gtagcgaaat cagcagccgg caactatgtc tttaacgaac gcaaaatgct
     1441 cgatgcaagc cacgtggtgg tcttttgtgc caaaaccgca atggacgatg tatggcttaa
     1501 gctggtagtg gaccaagagg atgcggatgg aaggttcgct acaccggagg cgaaagccgc
     1561 gaatgataag ggaagaaagt tctgggcaga catgcatcga aaagacctcc atgatgacgc
     1621 ggagtggatg gccaagcagg tttaccttaa tgtaggtaac ttcctgttgg gggtcgccgc
     1681 gttggggctg gacgctgtgc ctattgaagg ttttgacgcc gccattttgg atgcagagtt
     1741 tggactcaaa gaaaaaggct acactagcct cgtggtggtt ccggtcggtc accactcggt
     1801 ggaggacttt aacgcgaccc tccccaagtc gcggctgccg cagaatatca cactcacgga
     1861 agtggctggt tctagtgcag gtcaaggtgc acaggctggt caaggagcac aggctggttc
     1921 tagtgcaggt atggatatca tctcggtggc attgaaacgt cattctacca aagccttcga
     1981 tgcgtcgaaa aagcttacgc cagagcaagc tgaacagatc aagacgttgc ttcagtattc
     2041 acccagttca actaattccc aaccctggca cttcatcgtg gcgtccacgg aggaagggaa
     2101 agcgcgagta gcgaaatcag cagccggcaa ctatgtcttt aacgaacgca aaatgctcga
     2161 tgcaagccac gtggtggtct tttgtgccaa aaccgcaatg gacgatgtat ggcttaagct
     2221 ggtagtggac caagaggatg cggatggaag gttcgctaca ccggaggcga aagccgcgaa
     2281 tgataaggga agaaagttct gggcagacat gcatcgaaaa gacctccatg atgacgcgga
     2341 gtggatggcc aagcaggttt accttaatgt aggtaacttc ctgttggggg tcgccgcgtt
     2401 ggggctggac gctgtgccta ttgaaggttt tgacgccgcc attttggatg cagagtttgg
     2461 actcaaagaa aaaggctaca ctagcctcgt ggtggttccg gtcggtcacc actcggtgga
     2521 ggactttaac gcgaccctcc ccaagtcgcg gctgccgcag aatatcacac tcacggaagt
     2581 ggctggttct agtgcaggta tggtgagcaa gggcgaggag gataacatgg ccatcatcaa
     2641 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat
     2701 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     2761 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
     2821 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
     2881 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
     2941 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
     3001 cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg
     3061 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga
     3121 cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
     3181 gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac
     3241 catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct
     3301 gtacaaggct ggttctagct agaagcttat cgataatcaa cctctggatt acaaaatttg
     3361 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     3421 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3481 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     3541 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     3601 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     3661 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     3721 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     3781 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     3841 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     3901 ctccctttgg gccgcctccc cgcatcgata ccgtcgaccc gggcggccgc ttcgagcaga
     3961 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     4021 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     4081 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     4141 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag
     4201 agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag
     4261 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4321 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     4381 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     4441 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4501 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4561 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4621 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4681 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4741 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4801 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4861 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     4921 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4981 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5041 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5101 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5161 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5221 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5281 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5341 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5401 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5461 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5521 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5581 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5641 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5701 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5761 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5821 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5881 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5941 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6001 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6061 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6121 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6181 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6241 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6301 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6361 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6421 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6481 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6541 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6601 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6661 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6721 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     6781 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
