pAAV-hSyn-FLAG-iPAK4

LOCUS       pAAV-hSyn-FLAG-iPAK4                5661 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 5661)
  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 5661)
  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..5661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1058..2158
                     /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        592..1038
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     primer_bind     1010..1027
                     /label=syn-F
     CDS             1061..1084
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1094..1207
                     /codon_start=1
                     /label=INKA-box
                     /translation="EAEDWTAALLNRGRSRQPLVLGDNCFADLVHNWMELPE"
     CDS             1214..2155
                     /codon_start=1
                     /label=Pak4cat
                     /translation="PAAPAVPGPPGPRSPQREPQRVSHEQFRAALQLVVDPGDPRSYLD
                     NFIKIGEGSTGIVCIATVRSSGKLVAVKKMDLRKQQRRELLFNEVVIMRDYQHENVVEM
                     YNSYLVGDELWVVMEFLEGGALTDIVTHTRMNEEQIAAVCLAVLQALSVLHAQGVIHRD
                     IKSDSILLTHDGRVKLSDFGFCAQVSKEVPRRKSLVGTPYWMAPELISRLPYGPEVDIW
                     SLGIMVIEMVDGEPPYFNEPPLKAMKMIRDNLPPRLKNLHKVSPSLKGFLDRLLVRDPA
                     QRATAAELLKHPFLAKAGPPASIVPLMRQNRTR"
     primer_bind     complement(1307..1325)
                     /label=PAK4-R
     primer_bind     complement(2137..2159)
                     /label=NotI-PAK4-R
     primer_bind     complement(2138..2188)
                     /label=SalI-PAK4-R
     primer_bind     complement(2138..2180)
                     /label=HindIII-PAK4-R
     misc_feature    2171..2759
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2224..2244)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2642..2653)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2762..2778)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2763..2779)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2808..2929
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2819..2838)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2873..2892
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2911..2929)
                     /label=SV40-polyA
     repeat_region   3091..3220
                     /label=AAV2 ITR
     primer_bind     complement(3239..3256)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3239..3255)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3248..3270)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3397..3852
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3484..3503)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3694..3715
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3878..3982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3983..4843
                     /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(4201..4220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5014..5602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5503..5522
                     /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 ctgcagaggg ccctgcgtat gagtgcaagt
      601 gggttttagg accaggatga ggcggggtgg gggtgcctac ctgacgaccg accccaccca
      661 ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg
      721 ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct
      781 tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca
      841 ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc
      901 ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg
      961 cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg
     1021 tcgtgcctga gagcgcagtc gaattcgggg atccaccatg gactacaagg acgacgatga
     1081 taagtccgga tccgaagcag aggactggac agcagcccta ctgaacaggg gtcgcagtcg
     1141 ccagcccctg gtactagggg acaattgctt tgctgacttg gtgcacaact ggatggagct
     1201 gcctgaggaa ttccccgccg cccctgctgt tcctgggccc cctggccccc gctcaccaca
     1261 gcgggagcca cagcgagtat cccatgagca gttccgggct gccctgcagc tggtggtgga
     1321 cccaggcgac ccccgctcct acctggacaa cttcatcaag attggcgagg gctccacggg
     1381 catcgtgtgc atcgccaccg tgcgcagctc gggcaagctg gtggccgtca agaagatgga
     1441 cctgcgcaag cagcagaggc gcgagctgct cttcaacgag gtggtaatca tgagggacta
     1501 ccagcacgag aatgtggtgg agatgtacaa cagctacctg gtgggggacg agctctgggt
     1561 ggtcatggag ttcctggaag gaggcgccct caccgacatc gtcacccaca ccaggatgaa
     1621 cgaggagcag atcgcagccg tgtgccttgc agtgctgcag gccctgtcgg tgctccacgc
     1681 ccagggcgtc atccaccggg acatcaagag cgactcgatc ctgctgaccc atgatggcag
     1741 ggtgaagctg tcagactttg ggttctgcgc ccaggtgagc aaggaagtgc cccgaaggaa
     1801 gtcgctggtc ggcacgccct actggatggc cccagagctc atctcccgcc ttccctacgg
     1861 gccagaggta gacatctggt cgctggggat aatggtgatt gagatggtgg acggagagcc
     1921 cccctacttc aacgagccac ccctcaaagc catgaagatg attcgggaca acctgccacc
     1981 ccgactgaag aacctgcaca aggtgtcgcc atccctgaag ggcttcctgg accgcctgct
     2041 ggtgcgagac cctgcccagc gggccacggc agccgagctg ctgaagcacc cattcctggc
     2101 caaggcaggg ccgcctgcca gcatcgtgcc cctcatgcgc cagaaccgca ccagatgaaa
     2161 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2221 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2281 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2341 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2401 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2461 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2521 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     2581 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     2641 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     2701 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     2761 tcgataccgt cgacctcgac ccgggcggcc gcttcgagca gacatgataa gatacattga
     2821 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     2881 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     2941 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta
     3001 aaacctctac aaatgtggta aaatcgataa ggatcttcct agagcatggc tacgtagata
     3061 agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc
     3121 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     3181 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ccttaattaa cctaattcac
     3241 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     3301 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     3361 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     3421 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3481 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3541 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3601 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3661 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3721 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     3781 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     3841 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3901 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3961 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4021 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4081 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4141 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4201 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4261 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4321 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4381 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4441 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4501 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4561 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4621 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4681 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4741 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4801 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4861 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4921 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4981 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5041 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5101 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5161 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5221 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5281 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5341 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5401 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5461 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5521 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5581 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5641 ttgctggcct tttgctcaca t
//