LOCUS       sequence-243888-        7660 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NMNAT2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7660)
  AUTHORS   Ljungberg MC, Ali YO, Zhu J, Wu CS, Oka K, Zhai RG, Lu HC
  TITLE     CREB-activity and nmnat2 transcription are down-regulated prior to 
            neurodegeneration, while NMNAT2 over-expression is neuroprotective, 
            in a mouse model of human tauopathy.
  JOURNAL   Hum Mol Genet. 2012 Jan 15;21(2):251-67. doi: 10.1093/hmg/ddr492. 
            Epub 2011 Oct 25.
  PUBMED    22027994
REFERENCE   2  (bases 1 to 7660)
  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"; doi: 
            "10.1093/hmg/ddr492"; journalName: "Hum Mol Genet"; date: 
            "2012-01-15- 15"; volume: "21"; issue: "2"; pages: "251-67"
COMMENT     Sequence Label: AV4-EF1-NMNAT2
FEATURES             Location/Qualifiers
     source          1..7660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        187..1365
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          418..1356
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1313..1333
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1387..1413
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2353..2926
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2376..2928
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2520..2537)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2747..2766
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2946..2948,2949..2951,2952..3665)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2946..2948,2949..2951,2952..3662)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2991..3012)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3252..3271)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3599..3620
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3771..3790)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3836..3891
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3890..3909)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   4301..4430
                     /label=AAV2 ITR
     promoter        complement(4501..4519)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4502..4519)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4537..4553)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4537..4553)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4550..4572)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4561..4577
                     /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)."
     promoter        complement(join(4585..4591,4592..4609,4610..4615))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4630..4651
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4939..5527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5019..5038)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5698..6489,6490..6558))
                     /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     6321..6340
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6559..6663)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6741..7196)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6878..6899)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7090..7109
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7322..7344
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7336..7353
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7337..7353
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7360..7379
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7360..7378
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 acgtgagtgt tttgcgacat tttgcgacac catgtggtca cgctgggtat ttaagcccga
       61 gtgagcacgc agggtctcca tacgcgccct gcagtttccc aagctttgca aagatggata
      121 aagttttaaa cagagaggaa tctttgcagc taatggacct tctaggtctt gaaaggagtg
      181 ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
      241 ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg
      301 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
      361 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta
      421 agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct
      481 tgaattactt ccacctggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt
      541 gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag
      601 gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc
      661 gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt
      721 ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat ttcggttttt
      781 ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc
      841 ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg
      901 gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg
      961 gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagctcaaaa
     1021 tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc
     1081 tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac
     1141 ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga ggggttttat
     1201 gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg
     1261 atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct
     1321 cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaggaat tggatccacc
     1381 atggcatacc catatgatgt ccccgactac gcgaccgaga ccaccaagac ccacgttatc
     1441 ttgctcgcct gcggcagctt caatcccatc accaaagggc acattcagat gtttgaaaga
     1501 gccagggatt atctgcacaa aactggaagg tttattgtga ttggcgggat tgtctcccct
     1561 gtccacgact cctatggaaa acagggcctc gtgtcaagcc ggcaccgtct catcatgtgt
     1621 cagctggccg tccagaattc tgattggatc agggtggacc cttgggagtg ctaccaggac
     1681 acctggcaga cgacctgcag cgtgttggaa caccaccggg acctcatgaa gagggtgact
     1741 ggctgcatcc tctccaatgt caacacacct tccatgacac ctgtgatcgg acagccacaa
     1801 aacgagaccc cccagcccat ttaccagaac agcaacgtgg ccaccaagcc cactgcagcc
     1861 aagatcttgg ggaaggtggg agaaagcctc agccggatct gctgtgtccg cccgccggtg
     1921 gagcgtttca cctttgtaga tgagaatgcc aatctgggca cggtgatgcg gtatgaagag
     1981 attgagctac ggatcctgct gctgtgtggt agtgacctgc tggagtcctt ctgcatccca
     2041 gggctctgga acgaggcaga tatggaggtg attgttggtg actttgggat tgtggtggtg
     2101 ccccgggatg cagccgacac agaccgaatc atgaatcact cctcaatact ccgcaaatac
     2161 aaaaacaaca tcatggtggt gaaggatgac atcaaccatc ccatgtctgt tgtcagctca
     2221 accaagagca ggctggccct gcagcatggg gacggccatg ttgtggatta cctgtcccag
     2281 ccggtcatcg actacatcct caaaagccag ctgtacatca atgcctccgg ctaggcggcc
     2341 gcaattgatc cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga
     2401 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     2461 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     2521 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     2581 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     2641 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     2701 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     2761 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     2821 ggggcctcgg tacacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     2881 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaagat
     2941 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     3001 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     3061 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     3121 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     3181 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     3241 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     3301 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     3361 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     3421 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     3481 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3541 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     3601 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3661 agtaagaatt catcgatcta gagcatgcaa ttcactcctc aggtgcaggc tgcctatcag
     3721 aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc
     3781 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
     3841 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     3901 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     3961 acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat
     4021 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
     4081 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
     4141 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
     4201 cttctcttat ggagatccct cgacctgcag cccaagctta ggcgcgttct agagcatggc
     4261 tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag
     4321 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     4381 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ggttcgaaat
     4441 cgataagctt ggatcgatcc ggagagctcg tcgacggcgc gccatgcata gcttgagtat
     4501 tctatagtgt cacctaaata gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4561 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4621 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4681 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4741 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4801 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4861 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4921 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4981 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5041 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5101 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5161 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5221 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5281 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5341 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5401 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5461 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5521 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5581 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5641 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5701 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5761 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5821 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5881 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5941 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6001 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6061 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6121 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6181 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6241 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6301 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6361 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6421 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6481 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6541 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6601 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6661 gcgcacattt ccccgaaaag tgccacctga tgcggtgtga aataccgcac agatgcgtaa
     6721 ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     6781 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     6841 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     6901 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     6961 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     7021 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     7081 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     7141 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat
     7201 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     7261 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     7321 tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc
     7381 gaattgggcc cggcgcgccg acgcgcatgc tcctctagct agactcgagg aattcggtac
     7441 cccctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg gcctcagtga
     7501 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc ttgtagttaa
     7561 tgattaaccc gccatgctac ttatctacgt agccatgctc tagagtggag tcgtgacgtg
     7621 aattacgtca tagggttagg gaggtcctgt attagaggtc
//
