LOCUS       Exported                6026 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Viral expression of oScarlet.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-Ef1a-oScarlet
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6026)
  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 6026)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6026
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1432..2127
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWDRVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1565..1583)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1833..1852
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2153..2741
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2206..2226)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2624..2635)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2773..3249
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2972..2991)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3289..3429
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3289..3418
                     /label=AAV2 ITR
     rep_origin      3504..3959
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3591..3610)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3801..3822
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3976..3995)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4095..4117
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4155..4173)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4241..4345
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4346..5206
                     /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(4564..4583)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5377..5965
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5866..5885
                     /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 catggtgagc
     1441 aagggcgagg cagtgatcaa ggagttcatg cggttcaagg tgcacatgga gggctccatg
     1501 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     1561 accgccaagc tgaaggtgac caagggtggc cccctgccct tctcctggga catcctgtcc
     1621 cctcagttca tgtacggctc cagggccttc accaagcacc ccgccgacat ccccgactac
     1681 tataagcagt ccttccccga gggcttcaag tgggaccgcg tgatgaactt cgaggacggc
     1741 ggcgccgtga ccgtgaccca ggacacctcc ctggaggacg gcaccctgat ctacaaggtg
     1801 aagctccgcg gcaccaactt ccctcctgac ggccccgtaa tgcagaagaa gacaatgggc
     1861 tgggaagcgt ccaccgagcg gttgtacccc gaggacggcg tgctgaaggg cgacattaag
     1921 atggccctgc gcctgaagga cggcgggcgc tacctggcgg acttcaagac cacctacaag
     1981 gccaagaagc ccgtgcagat gcccggcgcc tacaacgtgg accgcaagtt ggacatcacc
     2041 tcccacaacg aggactacac cgtggtggaa cagtacgaac gctccgaggg ccgccactcc
     2101 accggcggca tggacgagct gtacaagtga attcgatatc aagcttatcg ataatcaacc
     2161 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     2221 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     2281 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     2341 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     2401 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     2461 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2521 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt
     2581 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     2641 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     2701 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct
     2761 cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa
     2821 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct
     2881 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa
     2941 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg
     3001 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag
     3061 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt
     3121 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg
     3181 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc
     3241 cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag
     3301 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     3361 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     3421 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3481 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3541 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3601 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3661 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     3721 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3781 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3841 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3901 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     3961 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4021 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4081 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4141 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4201 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4261 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4321 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4381 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4441 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4501 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4561 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4621 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4681 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4741 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4801 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4861 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4921 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4981 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5041 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5101 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5161 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5221 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5281 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5341 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5401 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5461 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5521 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5581 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5641 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5701 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5761 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5821 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5881 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5941 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6001 cttttgctgg ccttttgctc acatgt
//
