LOCUS       sequence-243919-        6706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control for AV4-EF1-NMNAT2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6706)
  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 6706)
  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-GFP
FEATURES             Location/Qualifiers
     source          1..6706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        80..1258
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          311..1249
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1206..1226
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    1292..1865
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1315..1867
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1459..1476)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1686..1705
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1885..1887,1888..1890,1891..2604)
                     /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(1885..1887,1888..1890,1891..2601)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1930..1951)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2191..2210)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2538..2559
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2710..2729)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2775..2830
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2829..2848)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   3240..3369
                     /label=AAV2 ITR
     promoter        complement(3440..3458)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3441..3458)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3476..3492)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3476..3492)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3489..3511)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3500..3516
                     /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(3524..3530,3531..3548,3549..3554))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3569..3590
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3878..4466)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3958..3977)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4637..5428,5429..5497))
                     /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     5260..5279
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5498..5602)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5680..6135)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5817..5838)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6029..6048
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6261..6283
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6275..6292
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6276..6292
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6299..6318
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6299..6317
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gcaaagatgg ataaagtttt aaacagagag gaatctttgc agctaatgga ccttctaggt
       61 cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac
      121 agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg
      181 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg
      241 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc
      301 cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg
      361 cccttgcgtg ccttgaatta cttccacctg gctgcagtac gtgattcttg atcccgagct
      421 tcgggttgga agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg
      481 tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct
      541 tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc
      601 tgcgacgctt tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg
      661 tatttcggtt tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc
      721 ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg
      781 ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag
      841 gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc
      901 agggagctca aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca
      961 aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc
     1021 gccgtccagg cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg
     1081 ggaggggttt tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc
     1141 agcttggcac ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt
     1201 cattctcaag cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgagg
     1261 aattggatcc cgggcggccg caattgatcc gcccctctcc ctcccccccc cctaacgtta
     1321 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca
     1381 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
     1441 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
     1501 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
     1561 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
     1621 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
     1681 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
     1741 attgtatggg atctgatctg gggcctcggt acacatgctt tacatgtgtt tagtcgaggt
     1801 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat
     1861 gataatatgg ccacaagatc caccatggtg agcaagggcg aggagctgtt caccggggtg
     1921 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1981 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2041 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     2101 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     2161 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     2221 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     2281 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     2341 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     2401 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     2461 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     2521 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     2581 ggcatggacg agctgtacaa gtaagaattc atcgatctag agcatgcaat tcactcctca
     2641 ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata
     2701 ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca
     2761 tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     2821 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta
     2881 tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct
     2941 ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa
     3001 agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac
     3061 atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct cctgactact
     3121 cccagtcata gctgtccctc ttctcttatg gagatccctc gacctgcagc ccaagcttag
     3181 gcgcgttcta gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa
     3241 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     3301 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     3361 agcgcgcagg gttcgaaatc gataagcttg gatcgatccg gagagctcgt cgacggcgcg
     3421 ccatgcatag cttgagtatt ctatagtgtc acctaaatag cttggcgtaa tcatggtcat
     3481 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3541 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3601 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3661 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3721 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3781 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3841 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3901 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3961 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4021 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4081 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4141 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4201 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4261 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4321 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4381 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4441 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4501 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4561 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4621 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4681 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4741 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4801 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4861 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4921 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4981 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5041 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5101 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5161 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5221 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5281 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5341 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5401 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5461 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5521 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5581 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgat gcggtgtgaa
     5641 ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt
     5701 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     5761 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     5821 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     5881 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     5941 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     6001 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     6061 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     6121 cgctacaggg cgcgtccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     6181 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     6241 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgta
     6301 atacgactca ctatagggcg aattgggccc ggcgcgccga cgcgcatgct cctctagcta
     6361 gactcgagga attcggtacc ccctgcgcgc tcgctcgctc actgaggccg cccgggcaaa
     6421 gcccgggcgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
     6481 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
     6541 agagtggagt cgtgacgtga attacgtcat agggttaggg aggtcctgta ttagaggtca
     6601 cgtgagtgtt ttgcgacatt ttgcgacacc atgtggtcac gctgggtatt taagcccgag
     6661 tgagcacgca gggtctccat acgcgccctg cagtttccca agcttt
//
