pAAV-cFos1-mChilada-iPAK4

LOCUS       pAAV-cFos1-mChilada-iPAK4             7560 bp DNA     circular SYN 23-APR-2024
DEFINITION  Express FlicR1 preferentially in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7560)
  AUTHORS   Abdelfattah AS, Farhi SL, Zhao Y, Brinks D, Zou P, Ruangkittisakul 
            A, Platisa J, Pieribone VA, Ballanyi K, Cohen AE, Campbell RE
  TITLE     A Bright and Fast Red Fluorescent Protein Voltage Indicator That 
            Reports Neuronal Activity in Organotypic Brain Slices.
  JOURNAL   J Neurosci. 2016 Feb 24;36(8):2458-72. doi: 
            10.1523/JNEUROSCI.3484-15.2016.
  PUBMED    26911693
REFERENCE   2  (bases 1 to 7560)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1523/JNEUROSCI.3484-15"; journalName: "J Neurosci"; date: 
            "2016-02-24- 24"; volume: "36"; issue: "8"; pages: "2458-72"
FEATURES             Location/Qualifiers
     source          1..7560
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          577..2253
                     /lab_host="."
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          2981..4057
                     /mol_type="genomic DNA"
                     /organism="."
     protein_bind    229..250
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        265..295
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    303..319
                     /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     308..330
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     327..343
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     327..343
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   381..510
                     /label=AAV2 ITR
     promoter        577..2253
                     /label=putative_fos
                     /note="/created_by=xiuyuanli
                     https://www.ncbi.nlm.nih.gov/nucleotide/V00727.1?report=gen
                     bank&log$=nuclalign&blast_rank=3&RID=55DC32WT016: 1-1618"
     promoter        873..1190
                     /label=putative_fos_from34856
                     /note="/created_by=xiuyuanli"
                     /note="/modified_by=xiuyuanli"
     misc_feature    1338..1345
                     /label=Fos oncogene start
     primer_bind     complement(2230..2275)
                     /label=MCS-cfos-R
     CDS             2273..2980
                     /codon_start=1
                     /label=mChilada
                     /translation="MVSKGEEDNMAIIKEYMRFKVHMEGSVNGHEFEIEGEGEGRPFEG
                     TQTAKLKVTKGGPLPFAWHILPPQFQYGSKAYVKHPADIPDYFKLSFPEGFTWEREMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGNTASTERMYPEDGALK
                     GETKWRLKLKDGGHYEAEVKTTYKAKKPVQLPGAYNVDRKLKITYHNEDYTIVEQYERA
                     EARHSTGGMDELYK"
     regulatory      2273..2276
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2960..3006
                     /label=GFP-iPAK4-F
     primer_bind     2960..3006
                     /label=Primer 1a
     CDS             2993..3106
                     /codon_start=1
                     /label=INKA-box
                     /translation="EAEDWTAALLNRGRSRQPLVLGDNCFADLVHNWMELPE"
     CDS             3113..4054
                     /codon_start=1
                     /label=Pak4cat
                     /translation="PAAPAVPGPPGPRSPQREPQRVSHEQFRAALQLVVDPGDPRSYLD
                     NFIKIGEGSTGIVCIATVRSSGKLVAVKKMDLRKQQRRELLFNEVVIMRDYQHENVVEM
                     YNSYLVGDELWVVMEFLEGGALTDIVTHTRMNEEQIAAVCLAVLQALSVLHAQGVIHRD
                     IKSDSILLTHDGRVKLSDFGFCAQVSKEVPRRKSLVGTPYWMAPELISRLPYGPEVDIW
                     SLGIMVIEMVDGEPPYFNEPPLKAMKMIRDNLPPRLKNLHKVSPSLKGFLDRLLVRDPA
                     QRATAAELLKHPFLAKAGPPASIVPLMRQNRTR"
     primer_bind     complement(3206..3224)
                     /label=PAK4-R
     primer_bind     complement(4037..4079)
                     /label=HindIII-PAK4-R
     misc_feature    4070..4658
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4123..4143)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4541..4552)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4661..4677)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4662..4678)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    4707..4828
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4718..4737)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4772..4791
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4810..4828)
                     /label=SV40-polyA
     repeat_region   4990..5119
                     /label=AAV2 ITR
     primer_bind     complement(5138..5155)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5138..5154)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5147..5169)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5296..5751
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5383..5402)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5593..5614
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5777..5881
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5882..6742
                     /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(6100..6119)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6913..7501
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7402..7421
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
       61 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
      121 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
      181 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
      241 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
      301 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccagatt
      361 taattaaggc cttaattagg ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc
      421 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
      481 gagtggccaa ctccatcact aggggttcct tgtagttaat gattaacccg ccatgctact
      541 tatctacgta gccatgctct aggaagatct ctgcagaagc tttcctttag gaacagaggc
      601 ttcgagcctt taaggctgcg tacttgcttc tcctaatacc agagactcaa aaaaaaaaaa
      661 aaagttccag attgctggac aatgacccgg gtctcatccc ttgaccctgg gaaccgggtc
      721 cacattgaat caggtgcgaa tgttcgctcg ccttctctgc ctttcccgcc tcccctcccc
      781 cggccgcggc cccggttccc cccctgcgct gcaccctcag agttggctgc agccggcgag
      841 ctgttcccgt caatccctcc ctcctttaca caggatgtcc atattaggac atctgcgtca
      901 gcaggtttcc acggccggtc cctgttgttc tggggggggg accatctccg aaatcctaca
      961 cgcggaaggt ctaggagacc ccctaagatc ccaaatgtga acactcatag gtgaaagatg
     1021 tatgccaaga cgggggttga aagcctgggg cgtagagttg acgacagagc gcccgcagag
     1081 ggccttgggg cgcgcttccc cccccttcca gttccgccca gtgacgtagg aagtccatcc
     1141 attcacagcg cttctataaa ggcgccagct gaggcgccta ctactccaac cgcgactgca
     1201 gcgagcaact gagaagactg gatagagccg gcggttccgc gaacgagcag tgaccgcgct
     1261 cccacccagc tctgctctgc agctcccacc agtgtctacc cctggacccc ttgccgggct
     1321 ttccccaaac ttcgaccatg atgttctcgg gtttcaacgc cgactacgag gcgtcatcct
     1381 cccgctgcag tagcgcctcc ccggccgggg acagcctttc ctactaccat tccccagccg
     1441 actccttctc cagcatgggc tctcctgtca acacacaggt gagtttggct ttgtgtagcc
     1501 gccaggtccg cgctgagggt cgccgtggag gagacactgg ggtgtgactc gcaggggcgg
     1561 gggggtcttc ctttttcgct ctggagggag actggcgcgg tcagagcagc cttagcctgg
     1621 gaacccagga cttgtctgag cgcgtgcaca cttgtcatag taagacttag tgaccccttc
     1681 ccgcgcggca ggtttattct gagtggcctg cctgcattct tctctcggcc gacttgtttc
     1741 tgagatcagc cggggccaac aagtctcgag caaagagtcg ctaactagag tttgggaggc
     1801 ggcaaaccgc ggcaatcccc cctcccgggg cagcctggag cagggaggag ggaggaggga
     1861 ggagggtgct gcgggcgggt gtgtaaggca gtttcattga taaaaagcga gttcattctg
     1921 gagactccgg agcagcgcct gcgtcagcgc agacgtcagg gatatttata acaaaccccc
     1981 tttcgagcga gtgatgccga agggataacg ggaacgcagc agtaggatgg aggggaaagg
     2041 ctgcgctgcg gaattcaagg gaggatattg ggagagcttt tatctccgat gaggtgcata
     2101 caggaagaca taagcagtct ctgaccggaa tgcttctctc tccctgcttc atgcgacact
     2161 agggccactt gctccacctg tgtctggaac ctcctcgctc acctccgctt tcctcttttt
     2221 gttttgtttc aggacttttg cgcagatctc tcctcgaatt cggggatcca ccatggtgag
     2281 caagggcgag gaggacaaca tggctatcat caaggaatac atgcgtttca aggttcatat
     2341 ggaaggttct gttaacggtc atgaattcga aatcgaaggt gaaggtgaag gtcgtccttt
     2401 cgaaggtacc cagaccgcta agctgaaggt taccaagggt ggtcctctgc ctttcgcttg
     2461 gcatatcctg cctcctcagt tccagtacgg ttctaaggct tacgttaagc atcctgctga
     2521 catccctgac tacttcaagc tgtctttccc tgaaggtttc acctgggaac gtgaaatgaa
     2581 cttcgaagac ggtggtgttg ttaccgttac ccaggactct tctctgcagg acggtgaatt
     2641 catctacaag gttaagctgc gtggtaccaa cttcccttct gacggtcctg ttatgcagaa
     2701 gaagaccatg ggtaacaccg cttctaccga acgtatgtac cctgaagacg gtgctctgaa
     2761 gggtgaaacc aagtggcgtc tgaagctgaa ggacggtggt cattacgaag ctgaagttaa
     2821 gaccacctac aaggctaaga agcctgttca gctgcctggt gcttacaacg ttgaccgtaa
     2881 gctgaagatc acctaccata acgaagacta caccatcgtt gaacagtacg aacgtgctga
     2941 agctcgtcat tctaccggtg gcatggacga gctgtacaag tcaggtggat ccgaagcaga
     3001 ggactggaca gcagccctac tgaacagggg tcgcagtcgc cagcccctgg tactagggga
     3061 caattgcttt gctgacttgg tgcacaactg gatggagctg cctgaggaat tccccgccgc
     3121 ccctgctgtt cctgggcccc ctggcccccg ctcaccacag cgggagccac agcgagtatc
     3181 ccatgagcag ttccgggctg ccctgcagct ggtggtggac ccaggcgacc cccgctccta
     3241 cctggacaac ttcatcaaga ttggcgaggg ctccacgggc atcgtgtgca tcgccaccgt
     3301 gcgcagctcg ggcaagctgg tggccgtcaa gaagatggac ctgcgcaagc agcagaggcg
     3361 cgagctgctc ttcaacgagg tggtaatcat gagggactac cagcacgaga atgtggtgga
     3421 gatgtacaac agctacctgg tgggggacga gctctgggtg gtcatggagt tcctggaagg
     3481 aggcgccctc accgacatcg tcacccacac caggatgaac gaggagcaga tcgcagccgt
     3541 gtgccttgca gtgctgcagg ccctgtcggt gctccacgcc cagggcgtca tccaccggga
     3601 catcaagagc gactcgatcc tgctgaccca tgatggcagg gtgaagctgt cagactttgg
     3661 gttctgcgcc caggtgagca aggaagtgcc ccgaaggaag tcgctggtcg gcacgcccta
     3721 ctggatggcc ccagagctca tctcccgcct tccctacggg ccagaggtag acatctggtc
     3781 gctggggata atggtgattg agatggtgga cggagagccc ccctacttca acgagccacc
     3841 cctcaaagcc atgaagatga ttcgggacaa cctgccaccc cgactgaaga acctgcacaa
     3901 ggtgtcgcca tccctgaagg gcttcctgga ccgcctgctg gtgcgagacc ctgcccagcg
     3961 ggccacggca gccgagctgc tgaagcaccc attcctggcc aaggcagggc cgcctgccag
     4021 catcgtgccc ctcatgcgcc agaaccgcac cagatgaaag cttatcgata atcaacctct
     4081 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     4141 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     4201 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     4261 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     4321 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     4381 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     4441 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     4501 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     4561 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     4621 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgacc
     4681 cgggcggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     4741 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     4801 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     4861 ggttcagggg gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa
     4921 aatcgataag gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca
     4981 ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     5041 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     5101 tgagcgagcg agcgcgcagc cttaattaac ctaattcact ggccgtcgtt ttacaacgtc
     5161 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     5221 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     5281 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5341 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     5401 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5461 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     5521 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5581 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     5641 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     5701 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     5761 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5821 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5881 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5941 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     6001 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     6061 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     6121 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     6181 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6241 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6301 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6361 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6421 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     6481 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6541 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6601 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6661 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6721 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6781 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6841 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6901 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6961 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     7021 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     7081 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     7141 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     7201 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     7261 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     7321 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7381 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7441 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     7501 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
//