pAAV-cFos2-mChilada-iPAK4

LOCUS       pAAV-cFos2-mChilada-iPAK4         6644 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 6644)
  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 6644)
  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..6644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          2065..3141
                     /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
     misc_feature    577..1337
                     /label=cfos-Luo liqun
     CDS             complement(1277..1288)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             1357..2064
                     /codon_start=1
                     /label=mChilada
                     /translation="MVSKGEEDNMAIIKEYMRFKVHMEGSVNGHEFEIEGEGEGRPFEG
                     TQTAKLKVTKGGPLPFAWHILPPQFQYGSKAYVKHPADIPDYFKLSFPEGFTWEREMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGNTASTERMYPEDGALK
                     GETKWRLKLKDGGHYEAEVKTTYKAKKPVQLPGAYNVDRKLKITYHNEDYTIVEQYERA
                     EARHSTGGMDELYK"
     regulatory      1357..1360
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2044..2090
                     /label=GFP-iPAK4-F
     primer_bind     2044..2090
                     /label=Primer 1a
     primer_bind     complement(2044..2090)
                     /label=XFP-iPAK4-R
     CDS             2077..2190
                     /codon_start=1
                     /label=INKA-box
                     /translation="EAEDWTAALLNRGRSRQPLVLGDNCFADLVHNWMELPE"
     CDS             2197..3138
                     /codon_start=1
                     /label=Pak4cat
                     /translation="PAAPAVPGPPGPRSPQREPQRVSHEQFRAALQLVVDPGDPRSYLD
                     NFIKIGEGSTGIVCIATVRSSGKLVAVKKMDLRKQQRRELLFNEVVIMRDYQHENVVEM
                     YNSYLVGDELWVVMEFLEGGALTDIVTHTRMNEEQIAAVCLAVLQALSVLHAQGVIHRD
                     IKSDSILLTHDGRVKLSDFGFCAQVSKEVPRRKSLVGTPYWMAPELISRLPYGPEVDIW
                     SLGIMVIEMVDGEPPYFNEPPLKAMKMIRDNLPPRLKNLHKVSPSLKGFLDRLLVRDPA
                     QRATAAELLKHPFLAKAGPPASIVPLMRQNRTR"
     primer_bind     complement(2290..2308)
                     /label=PAK4-R
     primer_bind     complement(3121..3163)
                     /label=HindIII-PAK4-R
     misc_feature    3154..3742
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3207..3227)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3625..3636)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3745..3761)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3746..3762)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3791..3912
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3802..3821)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3856..3875
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3894..3912)
                     /label=SV40-polyA
     repeat_region   4074..4203
                     /label=AAV2 ITR
     primer_bind     complement(4222..4239)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4222..4238)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4231..4253)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4380..4835
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4467..4486)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4677..4698
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4861..4965
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4966..5826
                     /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(5184..5203)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5997..6585
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6486..6505
                     /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 ttcgacctcg aattcgggga tccaccatgg tgagcaaggg cgaggaggac
     1381 aacatggcta tcatcaagga atacatgcgt ttcaaggttc atatggaagg ttctgttaac
     1441 ggtcatgaat tcgaaatcga aggtgaaggt gaaggtcgtc ctttcgaagg tacccagacc
     1501 gctaagctga aggttaccaa gggtggtcct ctgcctttcg cttggcatat cctgcctcct
     1561 cagttccagt acggttctaa ggcttacgtt aagcatcctg ctgacatccc tgactacttc
     1621 aagctgtctt tccctgaagg tttcacctgg gaacgtgaaa tgaacttcga agacggtggt
     1681 gttgttaccg ttacccagga ctcttctctg caggacggtg aattcatcta caaggttaag
     1741 ctgcgtggta ccaacttccc ttctgacggt cctgttatgc agaagaagac catgggtaac
     1801 accgcttcta ccgaacgtat gtaccctgaa gacggtgctc tgaagggtga aaccaagtgg
     1861 cgtctgaagc tgaaggacgg tggtcattac gaagctgaag ttaagaccac ctacaaggct
     1921 aagaagcctg ttcagctgcc tggtgcttac aacgttgacc gtaagctgaa gatcacctac
     1981 cataacgaag actacaccat cgttgaacag tacgaacgtg ctgaagctcg tcattctacc
     2041 ggtggcatgg acgagctgta caagtcaggt ggatccgaag cagaggactg gacagcagcc
     2101 ctactgaaca ggggtcgcag tcgccagccc ctggtactag gggacaattg ctttgctgac
     2161 ttggtgcaca actggatgga gctgcctgag gaattccccg ccgcccctgc tgttcctggg
     2221 ccccctggcc cccgctcacc acagcgggag ccacagcgag tatcccatga gcagttccgg
     2281 gctgccctgc agctggtggt ggacccaggc gacccccgct cctacctgga caacttcatc
     2341 aagattggcg agggctccac gggcatcgtg tgcatcgcca ccgtgcgcag ctcgggcaag
     2401 ctggtggccg tcaagaagat ggacctgcgc aagcagcaga ggcgcgagct gctcttcaac
     2461 gaggtggtaa tcatgaggga ctaccagcac gagaatgtgg tggagatgta caacagctac
     2521 ctggtggggg acgagctctg ggtggtcatg gagttcctgg aaggaggcgc cctcaccgac
     2581 atcgtcaccc acaccaggat gaacgaggag cagatcgcag ccgtgtgcct tgcagtgctg
     2641 caggccctgt cggtgctcca cgcccagggc gtcatccacc gggacatcaa gagcgactcg
     2701 atcctgctga cccatgatgg cagggtgaag ctgtcagact ttgggttctg cgcccaggtg
     2761 agcaaggaag tgccccgaag gaagtcgctg gtcggcacgc cctactggat ggccccagag
     2821 ctcatctccc gccttcccta cgggccagag gtagacatct ggtcgctggg gataatggtg
     2881 attgagatgg tggacggaga gcccccctac ttcaacgagc cacccctcaa agccatgaag
     2941 atgattcggg acaacctgcc accccgactg aagaacctgc acaaggtgtc gccatccctg
     3001 aagggcttcc tggaccgcct gctggtgcga gaccctgccc agcgggccac ggcagccgag
     3061 ctgctgaagc acccattcct ggccaaggca gggccgcctg ccagcatcgt gcccctcatg
     3121 cgccagaacc gcaccagatg aaagcttatc gataatcaac ctctggatta caaaatttgt
     3181 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3241 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3301 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     3361 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     3421 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     3481 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     3541 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     3601 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     3661 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     3721 tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gacccgggcg gccgcttcga
     3781 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3841 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     3901 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg
     3961 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatctt
     4021 cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc
     4081 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     4141 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     4201 cagccttaat taacctaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     4261 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     4321 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     4381 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4441 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4501 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4561 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4621 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4681 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4741 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4801 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     4861 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4921 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4981 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5041 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5101 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5161 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5221 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5281 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5341 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5401 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5461 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5521 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5581 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5641 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5701 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5761 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5821 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5881 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5941 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6001 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6061 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6121 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6181 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6241 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6301 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6361 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6421 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6481 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6541 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6601 gcctttttac ggttcctggc cttttgctgg ccttttgctc acat
//