LOCUS Exported 7711 bp ds-DNA circular SYN 11-FEB-2022 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pLV-mitoGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7711) AUTHORS Kitay BM, McCormack R, Wang Y, Tsoulfas P, Zhai RG TITLE Mislocalization of neuronal mitochondria reveals regulation of Wallerian degeneration and NMNAT/WLDS-mediated axon protection independent of axonal mitochondria. JOURNAL Hum Mol Genet. 2013 Jan 24. PUBMED 23314018 REFERENCE 2 (bases 1 to 7711) AUTHORS . TITLE Direct Submission JOURNAL Exported Feb 11, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7711 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..304 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 305..508 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 459..479 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 505..529 /label=LNCX /note="Human CMV promoter, forward primer" CDS 670..744 /codon_start=1 /product="mitochondrial presequence of human cytochrome c oxidase subunit VIII (Rizutto et al., 1989)" /label=COX8 presequence /translation="MSVLTPLLLRGLTGSARRLPVPRAK" CDS 808..1524 /codon_start=1 /product="Aequoria victoria green fluorescent protein" /label=GFP /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" primer_bind complement(838..866) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 1461..1481 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" primer_bind complement(1563..1579) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1564..1580) /label=pBluescriptKS /note="For pBluescript vector" misc_feature 1615..2203 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1668..1688) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2086..2097) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 2290..2523 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 2595..2716 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2632..2651) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2686..2705 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 2756..2891 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2818..2837 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(2911..2930) /label=T7 /note="T7 promoter, forward primer" promoter complement(2912..2930) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2940..2957) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2940..2956) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2949..2971) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3098..3553 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3185..3204) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3395..3416 /label=F1ori-F /note="F1 origin, forward primer" promoter 3579..3683 /gene="bla" /label=AmpR promoter CDS 3684..4544 /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(3902..3921) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4715..5303 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5204..5223 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5457..5474 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5591..5612 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5627..5657 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5665..5681 /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)." primer_bind 5670..5692 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5689..5705 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5689..5705 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 5724..5744 /label=T3 /note="T3 promoter, forward primer" promoter 5726..5744 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 5772..5998 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 5999..6179 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 6226..6351 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 6844..7077 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 7262..7306 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 7573..7689 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" ORIGIN 1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg 541 ccatccacgc tgttttgacc tccatagaag acaccgcccg ggcgtacggg atcccaccgg 601 tgctcgagat gcatactagt ggatcccccg ggctgcagga attcgataat tcgggtactc 661 cgtgccatca tgtccgtcct gacgccgctg ctgctgcggg gcttgacagg ctcggcccgg 721 cggctcccag tgccgcgcgc caagatccat tcgttgccgc cggaggggaa gcttatcgat 781 tatgatgttc ctgattatgc aagcttaatg agtaaaggag aagaactttt cactggagtt 841 gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga 901 gagggtgaag gtgatgcaac atacggaaaa cttaccctta aatttatttg cactactgga 961 aaactacctg ttccatggcc aacacttgtc actactttct cttatggtgt tcaatgcttt 1021 tcaagatacc cagatcatat gaaacggcat gactttttca agagtgccat gcccgaaggt 1081 tatgtacagg aaagaactat atttttcaaa gatgacggga actacaagac acgtgctgaa 1141 gtcaagtttg aaggtgatac ccttgttaat agaatcgagt taaaaggtat tgattttaaa 1201 gaagatggaa acattcttgg acacaaattg gaatacaact ataactcaca caatgtatac 1261 atcatggcag acaaacaaaa gaatggaatc aaagttaact tcaaaattag acacaacatt 1321 gaagatggaa gcgttcaact agcagatcat tatcaacaaa atactccaat tggcgatggc 1381 cctgtccttt taccagacaa ccattacctg tccacacaat ctgccctttc gaaagatccc 1441 aacgaaaaga gagaccacat ggtccttctt gagtttgtaa cagctgctgg gattacacat 1501 ggcatggatg aactatacaa ataaatgtcc agacttccgg ggatcctcta gatcaagctt 1561 atcgataccg tcgacctcga ctccggatga tcagggccct gtacagatat cgacaatcaa 1621 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt 1681 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct 1741 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc 1801 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg 1861 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc 1921 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc 1981 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt 2041 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca 2101 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt 2161 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat tcgagctcgg 2221 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa 2281 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta 2341 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc 2401 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt 2461 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta 2521 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat 2581 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 2641 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 2701 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc 2761 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 2821 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt 2881 ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca 2941 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc 3001 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc 3061 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta 3121 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg 3181 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa 3241 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc 3301 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt 3361 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca 3421 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc 3481 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta 3541 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat 3601 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc 3661 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct 3721 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag 3781 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta 3841 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc 3901 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca 3961 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg 4021 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg 4081 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca 4141 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa 4201 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa 4261 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata 4321 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat 4381 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc 4441 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata 4501 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt 4561 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga 4621 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag 4681 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa 4741 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag 4801 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg 4861 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat 4921 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta 4981 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg 5041 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc 5101 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa 5161 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc 5221 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt 5281 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct 5341 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc 5401 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg 5461 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt 5521 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc 5581 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc 5641 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct 5701 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc 5761 tgcaagctta atgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag 5821 ttagcaacat gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag 5881 gtggtacgat cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac 5941 cactgaattg ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg 6001 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 6061 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 6121 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 6181 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg 6241 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca 6301 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag 6361 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa 6421 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct 6481 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt 6541 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg 6601 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa 6661 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga 6721 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt 6781 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg 6841 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc 6901 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa 6961 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa 7021 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg 7081 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg 7141 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga 7201 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga 7261 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa 7321 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg 7381 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc 7441 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga 7501 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg 7561 gtatcggtta acttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat 7621 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaatt 7681 caaaatttta tcgataagct tgggagttcc g //