LOCUS       sequence-468739-        7089 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To Express a Rat NPAS4-DsRed fusion protein from a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7089)
  AUTHORS   Ploski JE, Monsey MS, Nguyen T, DiLeone RJ, Schafe GE
  TITLE     The neuronal PAS domain protein 4 (Npas4) is required for new and 
            reactivated fear memories.
  JOURNAL   PLoS One. 2011;6(8):e23760. doi: 10.1371/journal.pone.0023760. Epub 
            2011 Aug 22.
  PUBMED    21887312
REFERENCE   2  (bases 1 to 7089)
  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"; journalName: "PLoS One.";
            date: "2011"; volume: "6(8):e23760. doi"; pages: " 
            10.1371/journal.pone.0023760. Epub 2011 Aug 22"
COMMENT     Sequence Label: pDsRed1-CMV-Rat Npas4-DsRed1-pA
FEATURES             Location/Qualifiers
     source          1..7089
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..157
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    340..461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(377..396)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     431..450
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(468..923)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(605..626)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     817..836
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        950..1054
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1056..1413
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1056..1247
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1079..1099)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1264..1399
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1448..2242
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1448..2242
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1502..1521)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2112..2131
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2430..2449)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2474..2521
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2850..3438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3339..3358
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3613..3916
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3917..4120
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4071..4091
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6622..6631
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6628..6630,6631..6633,6634..7089,1..219)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(6811..6831)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     7029..7048
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 gacggcgtgc tgaagggcga gatccacaag gccctgaagc tgaaggacgg cggccactac
       61 ctggtggagt tcaagtccat ctacatggcc aagaagcccg tgcagctgcc cggctactac
      121 tacgtggact ccaagctgga catcacctcc cacaacgagg actacaccat cgtggagcag
      181 tacgagcgca ccgagggccg ccaccacctg ttcctgtagc ggccgcgact ctagatcata
      241 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
      301 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
      361 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
      421 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
      481 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
      541 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
      601 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
      661 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
      721 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
      781 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
      841 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
      901 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
      961 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     1021 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     1081 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     1141 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     1201 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     1261 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     1321 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     1381 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     1441 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     1501 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     1561 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     1621 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     1681 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     1741 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     1801 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     1861 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     1921 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     1981 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     2041 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     2101 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     2161 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     2221 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     2281 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     2341 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     2401 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     2461 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     2521 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     2581 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     2641 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     2701 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2761 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2821 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2881 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2941 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     3001 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3061 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3121 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3181 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3241 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3301 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3361 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3421 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3481 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3541 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     3601 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     3661 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     3721 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     3781 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     3841 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     3901 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     3961 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     4021 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     4081 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     4141 ccgctagcgc taccggactc agatctcgag atggaccgat ccaccaaggg cgcctccaag
     4201 gcgcgccgcg accagatcaa cgccgagatt cggaacctca aggaactgct gccgttggct
     4261 gaagcggaca aggtccggct gtcctacctg cacatcatga gtcttgcctg catctacact
     4321 cgcaagggtg tcttctttgc tggaggcact cctttggctg gccccacggg gcttctctct
     4381 gctcaagagc ttgaagacat agtggcagca ctacctggat ttctacttgt gttcacagct
     4441 gaggggaagt tgctatacct gtcggagagt gtgagcgagc atctgggcca ttctatggtg
     4501 gatctggttg cccagggtga cagtatttac gacatcattg accctgctga ccatctcact
     4561 gtgcgccagc agctcaccat gccctctgct ctggatgctg atcgcctttt ccgttgtcga
     4621 tttaacacat ccaagtccct ccggcgccag agtgcaggca acaaactggt gcttattcga
     4681 ggtcgattcc atgctcaccc acctggggcc tactgggcag gaaaccccgt gttcacagct
     4741 ttctgtgccc cactggagcc aagaccccgt cccggccctg gccctggccc tggccctggt
     4801 cctgcctctc tcttcctggc catgttccag agccggcatg ctaaggacct agccctactg
     4861 gacatttctg aaagtgtcct aatctacctg ggctttgagc gcagcgaact gctctgtaaa
     4921 tcatggtatg gactgctaca ccccgaggac ctggcccacg cttcttctca acactaccgc
     4981 ctgttggctg aaaatggaga tattcaggct gaaatggtgg tgagacttca agccaagcat
     5041 ggaggctgga catggattta ctgcatgcta tactcggatg gtccagaagg ccctattact
     5101 gccaataact accctatcag tgacacggaa gcctggagtc ttcgccagca gctaaactct
     5161 gaaaacaccc aggcagccta tgtcctagga accccagctg tgctaccctc attctctgag
     5221 aatgtcttct cccaggagca ctgctctaat ccactcttta caccagccct ggggactcct
     5281 agaagtgcca gcttccccag ggcccctgaa ctaggtgtga tctcaacatc agaagagctt
     5341 gcccaaccct ccaaagaact ggacttcagt tacctgccat tccctgcaag gcctgagcct
     5401 tccctccaag cagacttgag caaggatttg gtgtgtactc caccttacac accccaccag
     5461 ccaggaggct gcgccttcct cttcagcctc catgaaccct tccagactca cttgccccct
     5521 ccatccagct ctctccaaga acagctgacg ccaagcacgg tgactttctc tgaacagttg
     5581 acaccaagca gtgcaacctt cccagatcca ctaaccagtt cactacaagg acagttgact
     5641 gaaagctcag ccagaagctt tgaagaacaa ttgactccgt gcacctctac cttccctgac
     5701 cagctgcttc ccagcactgc cacgttccca gaacctctgg gtagccccac ccatgagcag
     5761 ctgactcctc ccagcacagc attccaagca catctgaaca gtcctagcca aaccttccca
     5821 gagcaactga gccctaatcc taccaagact tacttcgccc aggagggatg cagttttctc
     5881 tatgagaagt tgcccccaag tcctagcagc cctggtaatg gggactgtac actcttggcc
     5941 ctagctcaac tccggggtcc cctctctgtg gacgtccccc tggtgcctga aggcctgctc
     6001 acacctgagg cctctccagt caagcaaagt ttcttccact atacagagaa agagcagaat
     6061 gagatagatc gtctcatcca gcagatcagc cagttggctc agggcatgga caggcccttc
     6121 tcagctgagg ctggcactgg ggggctggag ccacttggag ggctggagcc cctgaacccc
     6181 aacctgtccc tgtcaggggc tggaccccct gtgcttagcc tggatcttaa accctggaaa
     6241 tgccaggagc tggacttctt ggttgaccct gataatttat tcctggaaga gacgccagtg
     6301 gaagacatct tcatggatct ttctactcca gaccccaatg gggaatgggg ttcaggggat
     6361 cctgaggcag aggtcccagg agggaccctg tcaccttgca acaacctgtc cccagaagat
     6421 cacagcttcc tggaggactt ggccacctat gaaaccgcct ttgagacagg tgtctcaaca
     6481 ttcccctatg aagggtttgc tgatgagttg catcaactcc agagccaagt tcaagacagc
     6541 ttccatgaag atggaagtgg aggggaacca acgtttcgaa ttctgcagtc gacggtaccg
     6601 cgggcccggg atccaccggt cgccaccatg gtgcgctcct ccaagaacgt catcaaggag
     6661 ttcatgcgct tcaaggtgcg catggagggc accgtgaacg gccacgagtt cgagatcgag
     6721 ggcgagggcg agggccgccc ctacgagggc cacaacaccg tgaagctgaa ggtgaccaag
     6781 ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttccagta cggctccaag
     6841 gtgtacgtga agcaccccgc cgacatcccc gactacaaga agctgtcctt ccccgagggc
     6901 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
     6961 tcctccctgc aggacggctg cttcatctac aaggtgaagt tcatcggcgt gaacttcccc
     7021 tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctccac cgagcgcctg
     7081 tacccccgc
//
