LOCUS       Exported                6181 bp ds-DNA     linear   SYN 23-MAY-2021
DEFINITION  AAV expression of Cre recombinase-activated jRGECO1b, a red 
            fluorescent calcium sensor protein, from the CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.CAG.Flex.NES-jRGECO1b.WPRE.SV40
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6181)
  AUTHORS   Dana H, Mohar B, Sun Y, Narayan S, Gordus A, Hasseman JP, Tsegaye G,
            Holt GT, Hu A, Walpita D, Patel R, Macklin JJ, Bargmann CI, Ahrens 
            MB, Schreiter ER, Jayaraman V, Looger LL, Svoboda K, Kim DS
  TITLE     Sensitive red protein calcium indicators for imaging neural 
            activity.
  JOURNAL   Elife. 2016 Mar 24;5. pii: e12727. doi: 10.7554/eLife.12727.
  PUBMED    27011354
REFERENCE   2  (bases 1 to 6181)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 23, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6181
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     251..273
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     335..354
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(389..422)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    complement(485..518)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     complement(996..1015)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(1711..1767)
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     CDS             complement(1795..1818)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             complement(1822..1854)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             complement(1858..1875)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    1958..1991
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    2062..2095
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2137..2725
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2190..2210)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2608..2619)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2768..2889
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2779..2798)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2833..2852
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3051..3180
                     /label=AAV2 ITR
     primer_bind     complement(3246..3263)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3417..4005)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3497..3516)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4176..5036)
                     /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     4799..4818
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5037..5141)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5167..5622)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5304..5325)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5516..5535
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5694..5823
                     /label=AAV2 ITR
ORIGIN
        1 tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc
       61 cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat
      121 gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg gagggccctt
      181 tgtgcggggg gagcggctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg
      241 ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat
      301 gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct
      361 catcattttg gcaaagaatt ggatccccat aacttcgtat aaagtatcct atacgaagtt
      421 atatcaaaat aggaagacca atgcttcacc atcgacccga attgccaagc atcaccatcg
      481 acccataact tcgtataatg tatgctatac gaagttatac tagagctttc acttcgctgt
      541 catcatttgt acaaactctt cgtagtttac ctgaccatcc ccatcgatgt ctgctaccct
      601 gatcatttca tcaacctcct catctgttaa cttctctcca aggtctgtca tcacgtggcg
      661 aagctctgct gcgccgatgt agccattgcc gtccttatca aacacgcgga acgcttcgcg
      721 aatttcctct tcactgtctg tgtcattcat ttttcttgcc atcatcgtca ggaactcagg
      781 gaagtcgaat gtgccgtcac cgtcggcatc tacttcattg atcatgtcct gcagctctgc
      841 ttctgtgggg ttctgcccca gcgatcgcat caccgtccca agctccttgg ttgttatcgt
      901 cccatccccg tccttgtcaa agcaggagaa agcctcttta aattctgcga tctgctcttc
      961 agtcagttgg tcacgcgtag cctcccagcc catggtcttc ttctgcatta cggggccgtc
     1021 cggagggaag ttggtgccgc gcagcttcac cttgtagatg aatacgccgt cctgcaggga
     1081 ggagtcctgg ttcacgtgaa taatgccgcc gtcctcgaag ttcatcacgc gctcccacct
     1141 gaagccctcg gggaaggaca gcttgaagta gtcggggatg tcggctgggt gcttaatgta
     1201 ggccttggag ccgtacatga actgagggga caggatgtcc caggcgaagg gcagggggcc
     1261 acccttggtc accttcagct tagcggtctg aaaggcctcg taggggcggc cctcgccctc
     1321 gccctcgatc tcgaactcgt ggccgttcac ggagccctcc atgtgcacct tgaagcgcat
     1381 gaactccttg atgatggcca tgttatcctc ctcgcccttg ctcaccagac tcccgcctgt
     1441 acctcccttg tacagctcgt ccatgccgcc ggtggagtgg cggccctcgg cacattcgca
     1501 ctgttccacg atggtgtagt cctcgttgtg ggagacgatg tccaacttga tgtccacctt
     1561 gtaggcgccg ggcagctgca cgggcttctt ggccttgtag gtagtcttga cctcggcggc
     1621 gtagtggccg ccgtccttca gcctcagccc cttcttgatc tcgctcttaa gggcgccgtc
     1681 ctcggggtac atccgctcgg aaaccacggg tgagctcagc cgacctatag ctctgactgc
     1741 gtgacctgcc ttattccact tacgacgtga tgagtcgacc attgttgcga gatccttatc
     1801 gtcatcgtcg tacagatccc gacccatttg ctgtccacca gtcatgctag ccataccatg
     1861 atgatgatga tgatgagaac ctcctccagt cttgttaata tcaagtccag ccaacttaag
     1921 agcaagctcg ttctgcagca tggtggcgga tcctagtata acttcgtata ggatacttta
     1981 tacgaagtta tcattgggat tcttcctatt ttgatccaag catcaccatc gaccctctag
     2041 tccagatctc accatcgacc cataacttcg tatagcatac attatacgaa gttatgtccc
     2101 tcgaagaggt tcgaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt
     2161 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     2221 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     2281 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     2341 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     2401 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     2461 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     2521 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     2581 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     2641 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     2701 atctcccttt gggccgcctc cccgcatcga taccgtcgac ccgggcggcc gcttcgagca
     2761 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     2821 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     2881 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
     2941 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatcttcct
     3001 agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct
     3061 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     3121 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     3181 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3241 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3301 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3361 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3421 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3481 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3541 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3601 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3661 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3721 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3781 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3841 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3901 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3961 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4021 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4081 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4141 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4201 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4261 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4321 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4381 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4441 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4501 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4561 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4621 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4681 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4741 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4801 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4861 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4921 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4981 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5041 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5101 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5161 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     5221 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     5281 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     5341 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     5401 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     5461 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     5521 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     5581 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     5641 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctgcgcg
     5701 ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc
     5761 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
     5821 ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc tctaggaaga
     5881 gtaccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     5941 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     6001 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     6061 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     6121 gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
     6181 t
//
