pAAV-cFos3-mChilada-iPAK4

LOCUS       pAAV-cFos3-mChilada-iPAK4              6201 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 6201)
  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 6201)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 16, 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..6201
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          577..894
                     /lab_host="."
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1622..2698
                     /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
     primer_bind     555..605
                     /label=PstI-cfos-shortF
     promoter        577..894
                     /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        577..894
                     /label=putative_fos_from34856
                     /note="/created_by=xiuyuanli"
                     /note="/modified_by=xiuyuanli"
     primer_bind     complement(872..916)
                     /label=MCS-shortCfos-R
     CDS             914..1621
                     /codon_start=1
                     /label=mChilada
                     /translation="MVSKGEEDNMAIIKEYMRFKVHMEGSVNGHEFEIEGEGEGRPFEG
                     TQTAKLKVTKGGPLPFAWHILPPQFQYGSKAYVKHPADIPDYFKLSFPEGFTWEREMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGNTASTERMYPEDGALK
                     GETKWRLKLKDGGHYEAEVKTTYKAKKPVQLPGAYNVDRKLKITYHNEDYTIVEQYERA
                     EARHSTGGMDELYK"
     regulatory      914..917
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1601..1647)
                     /label=XFP-iPAK4-R
     CDS             1634..1747
                     /codon_start=1
                     /label=INKA-box
                     /translation="EAEDWTAALLNRGRSRQPLVLGDNCFADLVHNWMELPE"
     CDS             1754..2695
                     /codon_start=1
                     /label=Pak4cat
                     /translation="PAAPAVPGPPGPRSPQREPQRVSHEQFRAALQLVVDPGDPRSYLD
                     NFIKIGEGSTGIVCIATVRSSGKLVAVKKMDLRKQQRRELLFNEVVIMRDYQHENVVEM
                     YNSYLVGDELWVVMEFLEGGALTDIVTHTRMNEEQIAAVCLAVLQALSVLHAQGVIHRD
                     IKSDSILLTHDGRVKLSDFGFCAQVSKEVPRRKSLVGTPYWMAPELISRLPYGPEVDIW
                     SLGIMVIEMVDGEPPYFNEPPLKAMKMIRDNLPPRLKNLHKVSPSLKGFLDRLLVRDPA
                     QRATAAELLKHPFLAKAGPPASIVPLMRQNRTR"
     primer_bind     complement(1847..1865)
                     /label=PAK4-R
     primer_bind     complement(2678..2720)
                     /label=HindIII-PAK4-R
     misc_feature    2711..3299
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2764..2784)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3182..3193)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3302..3318)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3303..3319)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3348..3469
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3359..3378)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3413..3432
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3451..3469)
                     /label=SV40-polyA
     primer_bind     complement(3572..3586)
                     /label=PstI-cfos-shortF
     repeat_region   3631..3760
                     /label=AAV2 ITR
     primer_bind     complement(3779..3796)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3779..3795)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3788..3810)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3937..4392
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4024..4043)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4234..4255
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4418..4522
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4523..5383
                     /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(4741..4760)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5554..6142
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6043..6062
                     /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 ctgcagggat gtccatatta ggacatctgc
      601 gtcagcaggt ttccacggcc ggtccctgtt gttctggggg ggggaccatc tccgaaatcc
      661 tacacgcgga aggtctagga gaccccctaa gatcccaaat gtgaacactc ataggtgaaa
      721 gatgtatgcc aagacggggg ttgaaagcct ggggcgtaga gttgacgaca gagcgcccgc
      781 agagggcctt ggggcgcgct tcccccccct tccagttccg cccagtgacg taggaagtcc
      841 atccattcac agcgcttcta taaaggcgcc agctgaggcg cctactactc caactcgaat
      901 tcggggatcc accatggtga gcaagggcga ggaggacaac atggctatca tcaaggaata
      961 catgcgtttc aaggttcata tggaaggttc tgttaacggt catgaattcg aaatcgaagg
     1021 tgaaggtgaa ggtcgtcctt tcgaaggtac ccagaccgct aagctgaagg ttaccaaggg
     1081 tggtcctctg cctttcgctt ggcatatcct gcctcctcag ttccagtacg gttctaaggc
     1141 ttacgttaag catcctgctg acatccctga ctacttcaag ctgtctttcc ctgaaggttt
     1201 cacctgggaa cgtgaaatga acttcgaaga cggtggtgtt gttaccgtta cccaggactc
     1261 ttctctgcag gacggtgaat tcatctacaa ggttaagctg cgtggtacca acttcccttc
     1321 tgacggtcct gttatgcaga agaagaccat gggtaacacc gcttctaccg aacgtatgta
     1381 ccctgaagac ggtgctctga agggtgaaac caagtggcgt ctgaagctga aggacggtgg
     1441 tcattacgaa gctgaagtta agaccaccta caaggctaag aagcctgttc agctgcctgg
     1501 tgcttacaac gttgaccgta agctgaagat cacctaccat aacgaagact acaccatcgt
     1561 tgaacagtac gaacgtgctg aagctcgtca ttctaccggt ggcatggacg agctgtacaa
     1621 gtcaggtgga tccgaagcag aggactggac agcagcccta ctgaacaggg gtcgcagtcg
     1681 ccagcccctg gtactagggg acaattgctt tgctgacttg gtgcacaact ggatggagct
     1741 gcctgaggaa ttccccgccg cccctgctgt tcctgggccc cctggccccc gctcaccaca
     1801 gcgggagcca cagcgagtat cccatgagca gttccgggct gccctgcagc tggtggtgga
     1861 cccaggcgac ccccgctcct acctggacaa cttcatcaag attggcgagg gctccacggg
     1921 catcgtgtgc atcgccaccg tgcgcagctc gggcaagctg gtggccgtca agaagatgga
     1981 cctgcgcaag cagcagaggc gcgagctgct cttcaacgag gtggtaatca tgagggacta
     2041 ccagcacgag aatgtggtgg agatgtacaa cagctacctg gtgggggacg agctctgggt
     2101 ggtcatggag ttcctggaag gaggcgccct caccgacatc gtcacccaca ccaggatgaa
     2161 cgaggagcag atcgcagccg tgtgccttgc agtgctgcag gccctgtcgg tgctccacgc
     2221 ccagggcgtc atccaccggg acatcaagag cgactcgatc ctgctgaccc atgatggcag
     2281 ggtgaagctg tcagactttg ggttctgcgc ccaggtgagc aaggaagtgc cccgaaggaa
     2341 gtcgctggtc ggcacgccct actggatggc cccagagctc atctcccgcc ttccctacgg
     2401 gccagaggta gacatctggt cgctggggat aatggtgatt gagatggtgg acggagagcc
     2461 cccctacttc aacgagccac ccctcaaagc catgaagatg attcgggaca acctgccacc
     2521 ccgactgaag aacctgcaca aggtgtcgcc atccctgaag ggcttcctgg accgcctgct
     2581 ggtgcgagac cctgcccagc gggccacggc agccgagctg ctgaagcacc cattcctggc
     2641 caaggcaggg ccgcctgcca gcatcgtgcc cctcatgcgc cagaaccgca ccagatgaaa
     2701 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2761 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2821 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2881 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2941 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     3001 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     3061 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     3121 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3181 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3241 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     3301 tcgataccgt cgacctcgac ccgggcggcc gcttcgagca gacatgataa gatacattga
     3361 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3421 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3481 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta
     3541 aaacctctac aaatgtggta aaatcgataa ggatcttcct agagcatggc tacgtagata
     3601 agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc
     3661 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     3721 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ccttaattaa cctaattcac
     3781 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     3841 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     3901 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     3961 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4021 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4081 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4141 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4201 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4261 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     4321 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4381 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4441 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4501 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4561 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4621 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4681 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4741 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4801 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4861 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4921 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4981 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5041 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5101 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5161 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5221 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5281 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5341 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5401 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5461 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5521 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5581 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5641 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5701 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5761 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5821 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5881 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5941 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6001 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6061 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6121 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6181 ttgctggcct tttgctcaca t
//