LOCUS       sequence-214713-        6838 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 6838)
  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 6838)
  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..6838
                     /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     1055..1077
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1139..1158
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2601..3308
                     /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(2746..2764)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3014..3033
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3337..3925
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3390..3410)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3808..3819)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3968..4089
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3979..3998)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4033..4052
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4251..4380
                     /label=AAV2 ITR
     rep_origin      4452..4907
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4539..4558)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4749..4770
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4933..5037
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5038..5106,5107..5898)
                     /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(5256..5275)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6069..6657
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6558..6577
                     /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 gatcgtacca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      301 ccattgacgt caatgggtgg actatttacg 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 cgctgcgacg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
      841 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
      901 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc
      961 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggctg tccgcggggg
     1021 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     1081 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     1141 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattggatcc taggaattct
     1201 gaactagtgc aggtatggat atcatctcgg tggcattgaa acgtcattct accaaagcct
     1261 tcgatgcgtc gaaaaagctt acgccagagc aagctgaaca gatcaagacg ttgcttcagt
     1321 attcacccag ttcaactaat tcccaaccct ggcacttcat cgtggcgtcc acggaggaag
     1381 ggaaagcgcg agtagcgaaa tcagcagccg gcaactatgt ctttaacgaa cgcaaaatgc
     1441 tcgatgcaag ccacgtggtg gtcttttgtg ccaaaaccgc aatggacgat gtatggctta
     1501 agctggtagt ggaccaagag gatgcggatg gaaggttcgc tacaccggag gcgaaagccg
     1561 cgaatgataa gggaagaaag ttctgggcag acatgcatcg aaaagacctc catgatgacg
     1621 cggagtggat ggccaagcag gtttacctta atgtaggtaa cttcctgttg ggggtcgccg
     1681 cgttggggct ggacgctgtg cctattgaag gttttgacgc cgccattttg gatgcagagt
     1741 ttggactcaa agaaaaaggc tacactagcc tcgtggtggt tccggtcggt caccactcgg
     1801 tggaggactt taacgcgacc ctccccaagt cgcggctgcc gcagaatatc acactcacgg
     1861 aagtggctgg ttctagtgca ggtcaaggtg cacaggctgg tcaaggagca caggctggtt
     1921 ctagtgcagg tatggatatc atctcggtgg cattgaaacg tcattctacc aaagccttcg
     1981 atgcgtcgaa aaagcttacg ccagagcaag ctgaacagat caagacgttg cttcagtatt
     2041 cacccagttc aactaattcc caaccctggc acttcatcgt ggcgtccacg gaggaaggga
     2101 aagcgcgagt agcgaaatca gcagccggca actatgtctt taacgaacgc aaaatgctcg
     2161 atgcaagcca cgtggtggtc ttttgtgcca aaaccgcaat ggacgatgta tggcttaagc
     2221 tggtagtgga ccaagaggat gcggatggaa ggttcgctac accggaggcg aaagccgcga
     2281 atgataaggg aagaaagttc tgggcagaca tgcatcgaaa agacctccat gatgacgcgg
     2341 agtggatggc caagcaggtt taccttaatg taggtaactt cctgttgggg gtcgccgcgt
     2401 tggggctgga cgctgtgcct attgaaggtt ttgacgccgc cattttggat gcagagtttg
     2461 gactcaaaga aaaaggctac actagcctcg tggtggttcc ggtcggtcac cactcggtgg
     2521 aggactttaa cgcgaccctc cccaagtcgc ggctgccgca gaatatcaca ctcacggaag
     2581 tggctggttc tagtgcaggt atggtgagca agggcgagga ggataacatg gccatcatca
     2641 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga
     2701 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     2761 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct
     2821 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg
     2881 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc
     2941 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
     3001 tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc
     3061 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg
     3121 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc
     3181 tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca
     3241 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
     3301 tgtacaaggc tggttctagc tagaagctta atcgataatc aacctctgga ttacaaaatt
     3361 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3421 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3481 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     3541 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     3601 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     3661 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     3721 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     3781 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     3841 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     3901 atctcccttt gggccgcctc cccgcatcga taccgtcgac ccgggcggcc gcttcgagca
     3961 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     4021 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     4081 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
     4141 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatcttcct
     4201 agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct
     4261 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     4321 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     4381 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     4441 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4501 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4561 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4621 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4681 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4741 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4801 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4861 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     4921 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4981 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5041 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5101 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5161 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5221 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5281 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5341 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5401 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5461 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5521 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5581 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5641 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5701 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5761 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5821 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5881 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5941 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6001 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6061 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6121 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6181 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6241 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6301 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6361 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6421 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6481 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6541 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6601 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6661 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6721 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     6781 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
