LOCUS       Exported                6573 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Viral expression of sRGECO.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-Ef1a-sRGECO
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6573)
  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 6573)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6573
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1416..1425
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1437..1493
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     2189..2208
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2700..3288
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2753..2773)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3171..3182)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3320..3796
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3519..3538)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3836..3976
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3836..3965
                     /label=AAV2 ITR
     rep_origin      4051..4506
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4138..4157)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4348..4369
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4523..4542)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4642..4664
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4702..4720)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4788..4892
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4893..5753
                     /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(5111..5130)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5924..6512
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6413..6432
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gatccgccac catggtggac tcatcacgtc
     1441 gtaagtggaa taaggcaggt cacgcagtca gagctatagg tcggctgagc tcacccgtgg
     1501 tttccgagcg gatgtacccc gaggacggcg ccctgaagag cgagatcaag aaggggctga
     1561 ggctgaagga cggcggccac tacgccgccg aggtcaagac cacctacaag gccaagaagc
     1621 ccgtgcagct gcccggcgcc tacatcgtgg acatcaagtt ggacatcgtg tcccacaacg
     1681 aggactacac catcgtggaa cagtgcgaac gcgccgaggg ccgccactcc accggcggca
     1741 tggacgagct gtacaaggga ggtacaggcg ggagtctggt gagcaagggc gaggaggata
     1801 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     1861 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgaggcc tttcagaccg
     1921 ctaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     1981 agttcatgta cggctccaag gcctacatta agcacccagc cgacatcccc gactacttca
     2041 agctgtcctt ccccgagggc ttcaggtggg accgcgtgat gaacttcgag gacggcggca
     2101 ttattcacgt gaaccaggac tcctccctgc aggacggcgt attcatctac aaggtgaagc
     2161 tgcgcggcac caacttcccc cccgacggcc ccgtaatgca gaagaagacc atgggctggg
     2221 aggctacgcg tgacgacctg actgaagagc agatcgcaga atttaaagag gctttctccc
     2281 tatttgacaa ggacggggat gggacgataa caaccaagga gcttgggacg gtgttccgat
     2341 cgctggggca gaaccccaca gaagcagagc tgcaggacat gatcaatgaa gtagatgccg
     2401 acggtgacgg cacattcgac ttccctgagt tcctgacgat gatggcaaga aaaatgaatg
     2461 acacagacag tgaagaggaa attcgcgaag cgttccgcgt gtttgataag gacggcaatg
     2521 gctacatcgg cgcagcagag cttcgccacg tgatgacaga ccttggagag aagttaacag
     2581 atgaggaggt tgatgaaatg atcagggtag cagacatcga tggggatggt caggtaaact
     2641 acgaagagtt tgtacaaatg atgacagcga agtaggaatt cgatatcaag cttatcgata
     2701 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2761 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     2821 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     2881 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     2941 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3001 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3061 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3121 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3181 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3241 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag
     3301 cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     3361 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     3421 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     3481 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     3541 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     3601 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     3661 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     3721 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     3781 tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa
     3841 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     3901 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     3961 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4021 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     4081 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     4141 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4201 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4261 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4321 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4381 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     4441 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     4501 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     4561 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4621 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4681 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     4741 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4801 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4861 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4921 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4981 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5041 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5101 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5161 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5221 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5281 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5341 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5401 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5461 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5521 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5581 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5641 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5701 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5761 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5821 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5881 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5941 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6001 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6061 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6121 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6181 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6241 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6301 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6361 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6421 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6481 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6541 tcctggcctt ttgctggcct tttgctcaca tgt
//
