pCAG-RFP-p2a-postASAP

LOCUS       pCAG-RFP-p2a-postASAP        7829 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses voltage sensor targeted to dendrites and spines along with
            RFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7829)
  AUTHORS   Cornejo VH, Ofer N, Yuste R
  TITLE     Voltage compartmentalization in dendritic spines in vivo.
  JOURNAL   Science. 2022 Jan 7;375(6576):82-86. doi: 10.1126/science.abg0501. 
            Epub 2021 Nov 11.
  PUBMED    34762487
REFERENCE   2  (bases 1 to 7829)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1126/science.abg0501"; journalName: "Science"; date: "2022-01-7-
            7"; volume: "375"; issue: "6576"; pages: "82-86"
COMMENT     Sequence Label: pCAG-RFP-p2a-postASAP
FEATURES             Location/Qualifiers
     source          1..7829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..660
                     /label=chicken beta-actin promoter
     intron          661..1677
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1601..1623
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1685..1704
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1765..1774
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1771..2481
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             2506..2562
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     regulatory      2839..2848
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    4150..4738
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4203..4223)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4621..4632)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4851..4870)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4916..4971
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4970..4989)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5332..5348)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5332..5348)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5345..5367)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5356..5372
                     /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)."
     promoter        complement(join(5380..5386,5387..5404,5405..5410))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5425..5446
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5505..5700
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5551..5686
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5706..5840
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5743..5762)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5797..5816
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5908..5925)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6079..6667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6159..6178)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6838..7629,7630..7698))
                     /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     7461..7480
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7699..7803)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg ggggcgcgcg ccaggcgggg cggggsgggg sgrggggsgg
      541 ggsggggsgr ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
      721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
      901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
      961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
     1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
     1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
     1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
     1261 gggccgcctc gggccggggr gggctcgggg grggggcgcg gcggcccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
     1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
     1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
     1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     1621 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1681 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt tctagagcgg
     1741 taccggatcc ccgggtaccg gtaggccacc atggtgtcta agggcgaaga gctgattaag
     1801 gagaacatgc acatgaagct gtacatggag ggcaccgtga acaaccacca cttcaagtgc
     1861 acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggtggtc
     1921 gagggcggcc ctctcccctt cgccttcgac atcctggcta ccagcttcat gtacggcagc
     1981 agaaccttca tcaaccacac ccagggcatc cccgacttct ttaagcagtc cttccctgag
     2041 ggcttcacat gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag
     2101 gacaccagcc tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc
     2161 ccatccaacg gccctgtgat gcagaagaaa acactcggct gggaggccaa caccgagatg
     2221 ctgtaccccg ctgacggcgg cctggaaggc agaagcgaca tggccctgaa gctcgtgggc
     2281 gggggccacc tgatctgcaa cttcaagacc acatacagat ccaagaaacc cgctaagaac
     2341 ctcaagatgc ccggcgtcta ctatgtggac cacagactgg aaagaatcaa ggaggccgac
     2401 aaagagacct acgtcgagca gcacgaggtg gctgtggcca gatactgcga cctccctagc
     2461 aaactggggc acaaacttaa tggcgcgccc gcgcccggct ccggagccac gaacttctct
     2521 ctgttaaagc aagcaggaga cgtggaagaa aaccccggtc ccatgctcga agtcaaggaa
     2581 gcatcaccaa ccagcatcca gatcagctgg gtgctccact tgcgccacgt tcgctactac
     2641 cgcatcacct acggtgaaac tggtggcaat agccctgtcc aggaattcac cgtgcctggc
     2701 agcaagtcca ctgctaccat cagcggcctg aaacctggtg tcgactatac catcacggtg
     2761 tacgccgtca cgatcttcag cgcctaccgc tccgcctggc cgccgatctc catcaactac
     2821 cgcaccggaa ccctagccgc caccatggag acgactgtga ggtatgaaca ggggtcagag
     2881 ctcactaaaa cttcgagctc tccaacagca gatgagccca cgataaagat tgatgatggt
     2941 cgtgatgagg gtaatgaaca agacagctgt tccaatacca ttaggagaaa aatttccccg
     3001 tttgtgatgt catttggatt cagagtattt ggagttgtgc ttatcattgt agacatcata
     3061 gtggtgattg tggatctggc catcagtgag aagaaaagag gcattagaga gattcttgaa
     3121 ggtgtttccc tggctatagc actcttcttc cttgttgatg ttctcatgag agtgtttgtt
     3181 gaaggcttca agaactattt ccggtccaaa ctgaatactt tggatgcagt catagtagtg
     3241 ggcactctgc taattaatat gacctactcc ttctctgacg gtacctttag gggcgataac
     3301 gtgtatatta ccgcggataa acagaaaaac ggcattaaag cgaactttac cgtgcgccat
     3361 aacgtggaag atggcagcgt gcagctggcg gatcattatc agcagaacac cccgattggc
     3421 gatggcccgg tgctgctgcc ggataaccat tatctgagca cccagaccgt gctgagcaaa
     3481 gatccgaacg aaaaacgcga tcacatggtg ctgctggaat ttgtgaccgc agcgggcatt
     3541 acacacggca tggatgaact gtatggcggc accggcggca gcgcgagcca gggcgaagaa
     3601 ctgtttaccg gcgtggtgcc gattctggtg gaactggatg gcgatgtgaa cggccataaa
     3661 tttagcgtgc gcggcgaagg cgaaggcgat gcgaccattg gcaaactgac cctgaaattt
     3721 atttgcacca ccggcaaact accggtgccg tggccgaccc tggtgaccac cttaacctat
     3781 ggcgtgcagt gctttagccg ctatccggat catatgaaac gccatgattt ttttaaaagc
     3841 gcgatgccgg aaggctatgt gcaggaacgc accattagct ttaaagatga tggcaaatat
     3901 aaaacccgcg cggtggtgaa atttgaaggc gataccctgg tgaaccgcat tgaactgaaa
     3961 ggcaccgatt ttaaagaaga tggcaacatt ctggggcata aactggaata taacacagat
     4021 cagatgccgc agatggttcg tcttcttcga gttctgagaa ttgttatctt aataagaata
     4081 tttcgcctgg cttcacagaa gaaacaactt gaagtggtaa cctaggaatt cgatatcaag
     4141 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     4201 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     4261 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     4321 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     4381 acccccactg gttggggaat tgccaccacc tgtcagctcc tttccggaac tttcgctttc
     4441 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     4501 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     4561 tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     4621 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     4681 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     4741 cgataccgag cgctgctcga gagatctggc cgcactcctc aggtgcaggc tgcctatcag
     4801 aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc
     4861 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
     4921 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     4981 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     5041 acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat
     5101 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
     5161 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
     5221 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
     5281 cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat ggtcatagct
     5341 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5401 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5461 actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca
     5521 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5581 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     5641 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5701 aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5761 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5821 tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5881 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5941 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6001 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6061 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6121 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6181 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6241 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6301 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6361 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6421 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6481 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6541 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6601 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6661 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6721 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6781 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6841 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6901 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6961 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7021 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7081 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7141 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7201 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7261 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7321 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7381 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7441 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7501 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7561 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7621 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7681 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7741 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7801 gcgcacattt ccccgaaaag tgccacctg
//