LOCUS       sequence-265091-        6583 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Viral expression of sRGECO.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6583)
  AUTHORS   Fenno LE, Ramakrishnan C, Kim YS, Evans KE, Lo M, Vesuna S, Inoue M,
            Cheung KYM, Yuen E, Pichamoorthy N, Hong ASO, Deisseroth K
  TITLE     Comprehensive Dual- and Triple-Feature Intersectional Single-Vector 
            Delivery of Diverse Functional Payloads to Cells of Behaving 
            Mammals.
  JOURNAL   Neuron. 2020 Jun 20. pii: S0896-6273(20)30432-3. doi: 
            10.1016/j.neuron.2020.06.003.
  PUBMED    32574559
REFERENCE   2  (bases 1 to 6583)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Neuron. 
            2020 Jun 20. pii: S0896-6273(20)30432-3. doi: 
            10.1016/j.neuron.2020.06.003."
COMMENT     Sequence Label: pAAV-Ef1a-sRGECO
FEATURES             Location/Qualifiers
     source          1..6583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1426..1435
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1447..1503
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     2199..2218
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2710..3298
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2763..2783)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3181..3192)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3330..3806
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3529..3548)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3846..3986
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3846..3975
                     /label=AAV2 ITR
     rep_origin      4061..4516
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4148..4167)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4358..4379
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4533..4552)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4652..4674
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4712..4730)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4798..4902
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4903..4971,4972..5763)
                     /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(5121..5140)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5934..6522
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6423..6442
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gatccgccac catggtggac
     1441 tcatcacgtc gtaagtggaa taaggcaggt cacgcagtca gagctatagg tcggctgagc
     1501 tcacccgtgg tttccgagcg gatgtacccc gaggacggcg ccctgaagag cgagatcaag
     1561 aaggggctga ggctgaagga cggcggccac tacgccgccg aggtcaagac cacctacaag
     1621 gccaagaagc ccgtgcagct gcccggcgcc tacatcgtgg acatcaagtt ggacatcgtg
     1681 tcccacaacg aggactacac catcgtggaa cagtgcgaac gcgccgaggg ccgccactcc
     1741 accggcggca tggacgagct gtacaaggga ggtacaggcg ggagtctggt gagcaagggc
     1801 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     1861 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgaggcc
     1921 tttcagaccg ctaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     1981 ctgtcccctc agttcatgta cggctccaag gcctacatta agcacccagc cgacatcccc
     2041 gactacttca agctgtcctt ccccgagggc ttcaggtggg accgcgtgat gaacttcgag
     2101 gacggcggca ttattcacgt gaaccaggac tcctccctgc aggacggcgt attcatctac
     2161 aaggtgaagc tgcgcggcac caacttcccc cccgacggcc ccgtaatgca gaagaagacc
     2221 atgggctggg aggctacgcg tgacgacctg actgaagagc agatcgcaga atttaaagag
     2281 gctttctccc tatttgacaa ggacggggat gggacgataa caaccaagga gcttgggacg
     2341 gtgttccgat cgctggggca gaaccccaca gaagcagagc tgcaggacat gatcaatgaa
     2401 gtagatgccg acggtgacgg cacattcgac ttccctgagt tcctgacgat gatggcaaga
     2461 aaaatgaatg acacagacag tgaagaggaa attcgcgaag cgttccgcgt gtttgataag
     2521 gacggcaatg gctacatcgg cgcagcagag cttcgccacg tgatgacaga ccttggagag
     2581 aagttaacag atgaggaggt tgatgaaatg atcagggtag cagacatcga tggggatggt
     2641 caggtaaact acgaagagtt tgtacaaatg atgacagcga agtaggaatt cgatatcaag
     2701 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2761 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2821 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2881 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2941 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     3001 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     3061 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     3121 tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3181 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3241 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     3301 cgataccgag cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc
     3361 ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa
     3421 gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg
     3481 tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac
     3541 caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag
     3601 cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca
     3661 gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca
     3721 actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt
     3781 gaaccactgc tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg
     3841 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     3901 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     3961 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     4021 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     4081 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4141 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4201 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4261 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4321 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4381 acaacactca accctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     4441 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4501 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4561 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     4621 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     4681 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4741 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4801 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4861 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4921 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4981 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5041 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5101 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5161 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5221 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5281 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5341 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5401 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5461 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5521 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5581 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5641 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5701 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5761 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5821 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5881 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5941 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6001 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6061 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     6121 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6181 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6241 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6301 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6361 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6421 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6481 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6541 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
