pAAV-NFkB-mGreenLantern-iPAK4

LOCUS       pAAV-NFkB-mGreenLantern-iPAK4             6176 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 6176)
  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 6176)
  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..6176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1597..2673
                     /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     549..607
                     /label=NFKB-F
     misc_feature    571..724
                     /label=Transcription blocker
                     /label=Transcription\blocker
     polyA_signal    571..619
                     /note="synthetic polyadenylation signal"
     misc_feature    633..724
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     misc_feature    758..767
                     /label=nfkb motif
     misc_feature    768..777
                     /label=nfkb motif
     misc_feature    778..787
                     /label=nfkb motif
     misc_feature    788..797
                     /label=nfkb motif
     promoter        810..841
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     misc_feature    811..841
                     /label=Minimal promoter
     misc_feature    817..823
                     /label=TATATAA
     primer_bind     859..897
                     /label=NFKB4-mGreenLantern-F
                     /note="for mScarlet3/I3 cloning"
     regulatory      874..883
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             880..1596
                     /codon_start=1
                     /label=mGreenLantern
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLGYGVACFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIVLKGIDFKEDGNILGHKLEYNFNSHKVYITADKQKNGIK
                     ANFKTRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLK
                     ERVTAAGITHDMDELYK"
     primer_bind     complement(1581..1622)
                     /label=INKA-mGreenLantern-R
     CDS             1609..1722
                     /codon_start=1
                     /label=INKA-box
                     /translation="EAEDWTAALLNRGRSRQPLVLGDNCFADLVHNWMELPE"
     CDS             1729..2670
                     /codon_start=1
                     /label=Pak4cat
                     /translation="PAAPAVPGPPGPRSPQREPQRVSHEQFRAALQLVVDPGDPRSYLD
                     NFIKIGEGSTGIVCIATVRSSGKLVAVKKMDLRKQQRRELLFNEVVIMRDYQHENVVEM
                     YNSYLVGDELWVVMEFLEGGALTDIVTHTRMNEEQIAAVCLAVLQALSVLHAQGVIHRD
                     IKSDSILLTHDGRVKLSDFGFCAQVSKEVPRRKSLVGTPYWMAPELISRLPYGPEVDIW
                     SLGIMVIEMVDGEPPYFNEPPLKAMKMIRDNLPPRLKNLHKVSPSLKGFLDRLLVRDPA
                     QRATAAELLKHPFLAKAGPPASIVPLMRQNRTR"
     primer_bind     complement(1822..1840)
                     /label=PAK4-R
     primer_bind     complement(2653..2695)
                     /label=HindIII-PAK4-R
     misc_feature    2686..3274
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2739..2759)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3157..3168)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3277..3293)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3278..3294)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3323..3444
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3334..3353)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3388..3407
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3426..3444)
                     /label=SV40-polyA
     primer_bind     complement(3547..3567)
                     /label=NFKB-F
     repeat_region   3606..3735
                     /label=AAV2 ITR
     primer_bind     complement(3754..3771)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3754..3770)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3763..3785)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3912..4367
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3999..4018)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4209..4230
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4393..4497
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4498..5358
                     /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(4716..4735)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5529..6117
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6018..6037
                     /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 aataaaatat ctttattttc attacatctg
      601 tgtgttggtt ttttgtgtga atcgatagta ctaacatacg ctctccatca aaacaaaacg
      661 aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt
      721 ctctatcgat aggtaccgag ctcttacgcg tgctagcggg aatttccggg aatttccggg
      781 aatttccggg aatttccaga tctagactct agagggtata taatggaagc tcgaattcca
      841 gcttggcatt ccggtactgt tggtaaaaag cttgccacca tggtgagcaa gggcgaggag
      901 ctgttcaccg gcgtggtgcc tatcctggtg gagctggacg gcgacgtgaa cggccacaag
      961 ttcagcgtga ggggcgaggg cgagggcgac gccaccaacg gcaagctgac cctgaagttc
     1021 atctgcacca ccggcaagct gcctgtgcct tggcctaccc tggtgaccac cctgggctac
     1081 ggcgtggcct gcttcgccag gtaccctgac cacatgaagc agcacgactt cttcaagagc
     1141 gccatgcctg agggctacgt gcaggagagg accatcagct tcaaggacga cggcacctac
     1201 aagaccaggg ccgaggtgaa gttcgagggc gacaccctgg tgaacaggat cgtgctgaag
     1261 ggcatcgact tcaaggagga cggcaacatc ctgggccaca agctggagta caacttcaac
     1321 agccacaagg tgtacatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     1381 accaggcaca acgtggagga cggcggcgtg cagctggccg accactacca gcagaacacc
     1441 cctatcggcg acggccctgt gctgctgcct gacaaccact acctgagcca ccagagcaag
     1501 ctgagcaagg accctaacga gaagagggac cacatggtgc tgaaggagag ggtgaccgcc
     1561 gccggcatca cccacgacat ggacgagctg tacaagtcag gtggatccga agcagaggac
     1621 tggacagcag ccctactgaa caggggtcgc agtcgccagc ccctggtact aggggacaat
     1681 tgctttgctg acttggtgca caactggatg gagctgcctg aggaattccc cgccgcccct
     1741 gctgttcctg ggccccctgg cccccgctca ccacagcggg agccacagcg agtatcccat
     1801 gagcagttcc gggctgccct gcagctggtg gtggacccag gcgacccccg ctcctacctg
     1861 gacaacttca tcaagattgg cgagggctcc acgggcatcg tgtgcatcgc caccgtgcgc
     1921 agctcgggca agctggtggc cgtcaagaag atggacctgc gcaagcagca gaggcgcgag
     1981 ctgctcttca acgaggtggt aatcatgagg gactaccagc acgagaatgt ggtggagatg
     2041 tacaacagct acctggtggg ggacgagctc tgggtggtca tggagttcct ggaaggaggc
     2101 gccctcaccg acatcgtcac ccacaccagg atgaacgagg agcagatcgc agccgtgtgc
     2161 cttgcagtgc tgcaggccct gtcggtgctc cacgcccagg gcgtcatcca ccgggacatc
     2221 aagagcgact cgatcctgct gacccatgat ggcagggtga agctgtcaga ctttgggttc
     2281 tgcgcccagg tgagcaagga agtgccccga aggaagtcgc tggtcggcac gccctactgg
     2341 atggccccag agctcatctc ccgccttccc tacgggccag aggtagacat ctggtcgctg
     2401 gggataatgg tgattgagat ggtggacgga gagcccccct acttcaacga gccacccctc
     2461 aaagccatga agatgattcg ggacaacctg ccaccccgac tgaagaacct gcacaaggtg
     2521 tcgccatccc tgaagggctt cctggaccgc ctgctggtgc gagaccctgc ccagcgggcc
     2581 acggcagccg agctgctgaa gcacccattc ctggccaagg cagggccgcc tgccagcatc
     2641 gtgcccctca tgcgccagaa ccgcaccaga tgaaagctta tcgataatca acctctggat
     2701 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     2761 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     2821 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     2881 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     2941 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     3001 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3061 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     3121 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     3181 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     3241 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgacccggg
     3301 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     3361 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     3421 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     3481 cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
     3541 gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa
     3601 ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     3661 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     3721 cgagcgagcg cgcagcctta attaacctaa ttcactggcc gtcgttttac aacgtcgtga
     3781 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     3841 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     3901 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3961 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4021 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4081 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4141 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4201 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4261 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4321 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     4381 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4441 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4501 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4561 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4621 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4681 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4741 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4801 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4861 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4921 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4981 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5041 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5101 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5161 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5221 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5281 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5341 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5401 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5461 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5521 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5581 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5641 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5701 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5761 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5821 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5881 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5941 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6001 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6061 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6121 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacat
//