LOCUS Exported 6836 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV-mediated expression of bacterial tdNfsB-mCherry (eNTR) under the CAG promoter. ACCESSION . VERSION . KEYWORDS aav-CAG-tdNfsB(F124W)-mCherry (eNTR) 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6836 /organism="synthetic DNA construct" /mol_type="other DNA" 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 5036..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" primer_bind 6809..6826 /label=L4440 /note="L4440 vector, 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 //