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" LTR 1..181 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 228..353 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 846..1079 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1264..1308 /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 1575..1691 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" enhancer 1714..2017 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2018..2221 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2172..2192 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2218..2242 /label=LNCX /note="Human CMV promoter, forward primer" CDS 2383..2457 /codon_start=1 /product="mitochondrial presequence of human cytochrome c oxidase subunit VIII (Rizutto et al., 1989)" /label=COX8 presequence /translation="MSVLTPLLLRGLTGSARRLPVPRAK" CDS 2521..3237 /codon_start=1 /product="Aequoria victoria green fluorescent protein" /label=GFP /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" primer_bind complement(2551..2579) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 3174..3194 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" primer_bind complement(3276..3292) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3277..3293) /label=pBluescriptKS /note="For pBluescript vector" misc_feature 3328..3916 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3381..3401) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3799..3810) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 4003..4236 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 4308..4429 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4345..4364) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4399..4418 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 4469..4604 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4531..4550 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(4624..4643) /label=T7 /note="T7 promoter, forward primer" promoter complement(4625..4643) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4653..4670) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4653..4669) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4662..4684) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 4811..5266 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4898..4917) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5108..5129 /label=F1ori-F /note="F1 origin, forward primer" promoter 5292..5396 /gene="bla" /label=AmpR promoter CDS 5397..6257 /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(5615..5634) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6428..7016 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6917..6936 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7170..7187 /label=L4440 /note="L4440 vector, forward primer" protein_bind 7304..7325 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 7340..7370 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7378..7394 /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 7383..7405 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7402..7418 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7402..7418 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 7437..7457 /label=T3 /note="T3 promoter, forward primer" promoter 7439..7457 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 7485..7711 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" ORIGIN 1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca 61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg 121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc 181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca 241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc 301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta 361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa 421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc 481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc 541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg 601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc 661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga 721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca 781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg 841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg 901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac 961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc 1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg 1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt 1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga 1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa 1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt 1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta 1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca 1441 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata 1501 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga 1561 cggtatcggt taacttttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga 1621 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa 1681 ttcaaaattt tatcgataag cttgggagtt ccgcgttaca taacttacgg taaatggccc 1741 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 1801 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 1861 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga 1921 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg 1981 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat 2041 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt 2101 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc 2161 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc 2221 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag 2281 aagacaccgc ccgggcgtac gggatcccac cggtgctcga gatgcatact agtggatccc 2341 ccgggctgca ggaattcgat aattcgggta ctccgtgcca tcatgtccgt cctgacgccg 2401 ctgctgctgc ggggcttgac aggctcggcc cggcggctcc cagtgccgcg cgccaagatc 2461 cattcgttgc cgccggaggg gaagcttatc gattatgatg ttcctgatta tgcaagctta 2521 atgagtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt 2581 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga 2641 aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt 2701 gtcactactt tctcttatgg tgttcaatgc ttttcaagat acccagatca tatgaaacgg 2761 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc 2821 aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt 2881 aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa 2941 ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca aaagaatgga 3001 atcaaagtta acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagat 3061 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac 3121 ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt 3181 cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata caaataaatg 3241 tccagacttc cggggatcct ctagatcaag cttatcgata ccgtcgacct cgactccgga 3301 tgatcagggc cctgtacaga tatcgacaat caacctctgg attacaaaat ttgtgaaaga 3361 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg 3421 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc 3481 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc 3541 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt 3601 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt 3661 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg 3721 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg 3781 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg 3841 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt 3901 tgggccgcct ccccgcctgg aattcgagct cggtaccttt aagaccaatg acttacaagg 3961 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact 4021 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct 4081 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc 4141 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc 4201 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta 4261 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg 4321 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 4381 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc 4441 tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca 4501 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 4561 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat 4621 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac 4681 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc 4741 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat 4801 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 4861 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 4921 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg 4981 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca 5041 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc 5101 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct 5161 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa 5221 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc 5281 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc 5341 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga 5401 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt 5461 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag 5521 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag 5581 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta 5641 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg 5701 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca 5761 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag 5821 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc 5881 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg 5941 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc 6001 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg 6061 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg 6121 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga 6181 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac 6241 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa 6301 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca 6361 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag 6421 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 6481 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 6541 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc 6601 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 6661 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 6721 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 6781 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc 6841 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 6901 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 6961 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 7021 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct 7081 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata 7141 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc 7201 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg 7261 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca 7321 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg 7381 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa 7441 ttaaccctca ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt cttatgcaat 7501 actcttgtag tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa 7561 aaagcaccgt gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag 7621 gcaacagacg ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat 7681 tgtatttaag tgcctagctc gatacataaa c //