LOCUS       sequence-354363-        7711 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Hum Mol 
            Genet. 2013 Jan 24."
COMMENT     Sequence Label: pLV-mitoGFP
FEATURES             Location/Qualifiers
     source          1..7711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             781..807
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             808..1524
                     /codon_start=1
                     /product="Aequorea 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     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             join(3684..3752,3753..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        join(5627..5632,5633..5650,5651..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
//
