LOCUS       Exported                6406 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Viral expression of ChRmine-mScarlet.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-nEF-ChRmine-mScarlet
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6406)
  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 6406)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6406
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        177..388
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             401..669
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      709..718
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1729..1785
                     /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"
     CDS             1786..2481
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1919..1937)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2187..2206
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2533..3121
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2586..2606)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3004..3015)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3153..3629
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3352..3371)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3669..3809
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3669..3798
                     /label=AAV2 ITR
     rep_origin      3884..4339
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3971..3990)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4181..4202
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4356..4375)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4475..4497
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4535..4553)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4621..4725
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4726..5586
                     /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(4944..4963)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5757..6345
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6246..6265
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgmcckwy
       61 gsycgggcgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaaggat ctgcgatcgc tccggtgccc gtcagtgggc
      181 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg
      241 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
      301 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
      361 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc
      421 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc
      481 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc
      541 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac
      601 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg
      661 ttacagatcc aagctgtgac cggcgcctac ggatccccgg gtaccggtgc caccatggca
      721 cacgcaccag gcaccgacca gatgttctac gtgggcacaa tggacggctg gtatctggat
      781 accaagctga actccgtggc catcggcgcc cactggtctt gctttatcgt gctgacaatc
      841 accacattct acctgggcta tgagtcctgg acatctcggg gccctagcaa gagaacctcc
      901 ttttacgccg gctatcagga ggagcagaac ctggccctgt ttgtgaattt ctttgccatg
      961 ctgagctact tcggcaagat cgtggccgac acactgggcc acaacttcgg cgatgtgggc
     1021 cccttcatca tcggcttcgg caattacagg tatgccgact acatgctgac ctgcccaatg
     1081 ctggtgtacg atctgctgta tcagctgagg gccccctatc gcgtgtcttg tagcgccatc
     1141 atctttgcca tcctgatgtc tggcgtgctg gccgagttct acgcagaggg cgaccctagg
     1201 ctgaggaatg gcgcctacgc ctggtatggc tttggctgtt tctggtttat cttcgcctac
     1261 tctatcgtga tgagcatcgt ggccaagcag tatagccggc tggcccagct ggcccaggat
     1321 acaggcgccg agcacagcct gcacgtgctg aagttcgccg tgtttacctt ctccatgctg
     1381 tggattctgt ttcccctggt gtgggccatc tgccctagag gcttcggctg gatcgacgat
     1441 aactggacag aggtggccca ctgcgtgtgc gacatcgtgg ccaagtcctg ttacggcttt
     1501 gccctggccc ggttcagaaa gacctatgat gaggagctgt ttcggctgct ggagcagctg
     1561 ggacacgacg aggatgagtt ccagaagctg gagctggata tgaggctgag cagcaatggc
     1621 gagcgcctgc ggagactgtc tgcggccgcc aagagcagga tcaccagcga gggcgagtac
     1681 atccccctgg accagatcga catcaacgtg ggcgcgcccg gctccggagc cacgaacttc
     1741 tctctgttaa agcaagcagg agacgtggaa gaaaaccccg gtcccatggt gagcaagggc
     1801 gaggcagtga tcaaggagtt catgcggttc aaggtgcaca tggagggctc catgaacggc
     1861 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     1921 aagctgaagg tgaccaaggg tggccccctg cccttctcct gggacatcct gtcccctcag
     1981 ttcatgtacg gctccagggc cttcaccaag caccccgccg acatccccga ctactataag
     2041 cagtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgcc
     2101 gtgaccgtga cccaggacac ctccctggag gacggcaccc tgatctacaa ggtgaagctc
     2161 cgcggcacca acttccctcc tgacggcccc gtaatgcaga agaagacaat gggctgggaa
     2221 gcgtccaccg agcggttgta ccccgaggac ggcgtgctga agggcgacat taagatggcc
     2281 ctgcgcctga aggacggcgg gcgctacctg gcggacttca agaccaccta caaggccaag
     2341 aagcccgtgc agatgcccgg cgcctacaac gtggaccgca agttggacat cacctcccac
     2401 aacgaggact acaccgtggt ggaacagtac gaacgctccg agggccgcca ctccaccggc
     2461 ggcatggacg agctgtacaa gttctgctac gagaacgagg tgtaatgaga attcgatatc
     2521 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2581 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2641 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     2701 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     2761 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     2821 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     2881 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     2941 ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     3001 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     3061 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     3121 catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac ccctccccag
     3181 tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat
     3241 taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg
     3301 tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg
     3361 aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc
     3421 aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc
     3481 tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct
     3541 ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg
     3601 cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg tgcggaccga
     3661 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     3721 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     3781 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     3841 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     3901 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3961 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4021 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4081 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4141 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4201 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     4261 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     4321 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4381 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4441 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4501 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     4561 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     4621 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4681 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4741 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4801 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4861 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4921 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4981 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5041 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5101 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5161 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5221 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5281 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5341 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5401 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5461 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5521 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5581 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5641 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5701 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5761 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5821 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5881 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5941 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6001 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6061 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6121 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6181 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6241 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6301 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6361 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
