LOCUS       sequence-227800-        6581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mouse C-Fos.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6581)
  AUTHORS   Pellegrino MJ, Stork PJ
  TITLE     Sustained activation of extracellular signal-regulated kinase by 
            nerve growth factor regulates c-fos protein stabilization and 
            transactivation in PC12 cells.
  JOURNAL   J Neurochem. 2006 Dec;99(6):1480-93. doi: 
            10.1111/j.1471-4159.2006.04250.x.
  PUBMED    17223854
REFERENCE   2  (bases 1 to 6581)
  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: 
            "10.1111/j.1471-4159.2006.04250"; journalName: "J Neurochem"; date: 
            "2006-12"; volume: "99"; issue: "6"; pages: "1480-93"
COMMENT     Sequence Label: Flag-Fos S374A
FEATURES             Location/Qualifiers
     source          1..6581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        107..486
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        487..690
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     641..661
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     735..754
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        735..753
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      785..794
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             794..841
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     promoter        complement(2006..2024)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2007..2024)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2044..2061)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2050..2274
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2320..2748
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2407..2426)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2617..2638
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2757..2777)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2762..3091
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2942..3077
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3004..3023
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3158..3952
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3212..3231)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3822..3841
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4126..4247
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4163..4182)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4217..4236
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4296..4312)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4296..4312)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4309..4331)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4320..4336
                     /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(4344..4350,4351..4368,4369..4374))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4389..4410
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4527..4544)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4698..5286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4778..4797)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5457..6248,6249..6317))
                     /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     6080..6099
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6318..6422)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6497..6516)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
       61 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
      121 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      181 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      241 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      301 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      361 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      421 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      481 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      541 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      601 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      661 gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac
      721 tggcttatcg aaattaatac gactcactat agggagaccc aagcttggta ccgagctcgg
      781 atccgccacc atggactaca aggacgacga tgacaaggac tacaaggacg acgatgacaa
      841 gaattctatg atgttctcgg gtttcaacgc cgactacgag gcgtcatcct cccgctgcag
      901 tagcgcctcc ccggccgggg acagcctttc ctactaccat tccccagccg actccttctc
      961 cagcatgggc tctcctgtca acacacagga cttttgcgca gatctgtccg tctctagtgc
     1021 caactttatc cccacggtga cagccatctc caccagccca gacctgcagt ggctggtgca
     1081 gcccactctg gtctcctccg tggccccatc gcagaccaga gcgccccatc cttacggact
     1141 ccccacccag tctgctgggg cttacgccag agcgggaatg gtgaagaccg tgtcaggagg
     1201 cagagcgcag agcatcggca gaaggggcaa agtagagcag ctatctcctg aagaggaaga
     1261 gaaacggaga atccgaaggg aacggaataa gatggctgca gccaagtgcc ggaatcggag
     1321 gagggagctg acagatacac tccaagcgga gacagatcaa cttgaagatg agaagtctgc
     1381 gttgcagact gagattgcca atctgctgaa agagaaggaa aaactggagt ttattttggc
     1441 agcccaccga cctgcctgca agatccccga tgaccttggc ttcccagagg agatgtctgt
     1501 ggcctccctg gatttgactg gaggtctgcc tgaggcttcc accccagagt ctgaggaggc
     1561 cttcaccctg ccccttctca acgaccctga gcccaagcca tccttggagc cagtcaagag
     1621 catcagcaac gtggagctga aggcagaacc ctttgatgac ttcttgtttc cggcatcatc
     1681 taggcccagt ggctcagaga cctcccgctc tgtgccagat gtggacctgt ccggttcctt
     1741 ctatgcagca gactgggagc ctctgcacag caattccttg gggatggggc ccatggtcac
     1801 agagctggag cccctgtgta ctcccgtggt cacctgtact ccgggctgca ctacttacac
     1861 gtcttccttt gtcttcacct accctgaagc tgactccttc ccaagctgtg ccgctgccca
     1921 ccgaaagggc agcagcagca acgagccctc ctccgactcc ctgagcgcac ccacgctgct
     1981 ggccctgtga tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat
     2041 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2101 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2161 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2221 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     2281 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     2341 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2401 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2461 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2521 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2581 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2641 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     2701 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     2761 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2821 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2881 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2941 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3001 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     3061 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     3121 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3181 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3241 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3301 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     3361 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3421 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3481 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3541 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     3601 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     3661 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     3721 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     3781 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     3841 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     3901 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     3961 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4021 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4081 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4141 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4201 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4261 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4321 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4381 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4441 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4501 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4561 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4621 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4681 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4741 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4801 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4861 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4921 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4981 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5041 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5101 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5161 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5221 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5281 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5341 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5401 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5461 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5521 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5581 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5641 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5701 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5761 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5821 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5881 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5941 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6001 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6061 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6121 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6181 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6241 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6301 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6361 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6421 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     6481 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     6541 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa a
//
