LOCUS       Exported                6402 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV expression of Cre recombinase-activated jRGECO1b, a red 
            fluorescent calcium sensor protein, from the Synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.Syn.Flex.NES-jRGECO1b.WPRE.SV40
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6402)
  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 6402)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6402
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   222..351
                     /label=AAV2 ITR
     polyA_signal    513..634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(550..569)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     604..623
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    677..1265
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             783..794
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1192..1212
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    complement(1307..1340)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    complement(1411..1444)
                     /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)."
     CDS             1527..1544
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1548..1580
                     /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             1584..1607
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1635..1691
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     2387..2406
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    2884..2917
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    2980..3013
                     /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)."
     promoter        complement(3043..3490)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   3572..3701
                     /label=AAV2 ITR
     rep_origin      3773..4228
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3860..3879)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4070..4091
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4254..4358
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4359..5219
                     /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(4577..4596)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5390..5978
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5879..5898
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6266..6287
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6302..6332
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6340..6356
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6345..6367
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6364..6380
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6364..6380
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
       61 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
      121 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
      181 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctgcgcgct cgctcgctca
      241 ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
      301 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc ttgtagttaa
      361 tgattaaccc gccatgctac ttatctacgt agccatgctc taggaagatc cttatcgatt
      421 ttaccacatt tgtagaggtt ttacttgctt taaaaaacct cccacatctc cccctgaacc
      481 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
      541 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
      601 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgctcgaag cggccgcccg
      661 ggtcgacggt atcgatgcgg ggaggcggcc caaagggaga tccgactcgt ctgagggcga
      721 aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg aaggtccgct
      781 ggattgaggg ccgaagggac gtagcagaag gacgtcccgc gcagaatcca ggtggcaaca
      841 caggcgagca gccaaggaaa ggacgatgat ttccccgaca acaccacgga attgtcagtg
      901 cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg
      961 gcaataggga gggggaaagc gaaagtcccg gaaaggagct gacaggtggt ggcaatgccc
     1021 caaccagtgg gggttgcgtc agcaaacaca gtgcacacca cgccacgttg cctgacaacg
     1081 ggccacaact cctcataaag agacagcaac caggatttat acaaggagga gaaaatgaaa
     1141 gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc acatagcgta
     1201 aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta atccagaggt
     1261 tgattatcga taagcttgat atcgaattcg aacctcttcg agggacataa cttcgtataa
     1321 tgtatgctat acgaagttat gggtcgatgg tgagatctgg actagagggt cgatggtgat
     1381 gcttggatca aaataggaag aatcccaatg ataacttcgt ataaagtatc ctatacgaag
     1441 ttatactagg atccgccacc atgctgcaga acgagcttgc tcttaagttg gctggacttg
     1501 atattaacaa gactggagga ggttctcatc atcatcatca tcatggtatg gctagcatga
     1561 ctggtggaca gcaaatgggt cgggatctgt acgacgatga cgataaggat ctcgcaacaa
     1621 tggtcgactc atcacgtcgt aagtggaata aggcaggtca cgcagtcaga gctataggtc
     1681 ggctgagctc acccgtggtt tccgagcgga tgtaccccga ggacggcgcc cttaagagcg
     1741 agatcaagaa ggggctgagg ctgaaggacg gcggccacta cgccgccgag gtcaagacta
     1801 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caaggtggac atcaagttgg
     1861 acatcgtctc ccacaacgag gactacacca tcgtggaaca gtgcgaatgt gccgagggcc
     1921 gccactccac cggcggcatg gacgagctgt acaagggagg tacaggcggg agtctggtga
     1981 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     2041 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     2101 acgaggcctt tcagaccgct aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     2161 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacattaag cacccagccg
     2221 acatccccga ctacttcaag ctgtccttcc ccgagggctt caggtgggag cgcgtgatga
     2281 acttcgagga cggcggcatt attcacgtga accaggactc ctccctgcag gacggcgtat
     2341 tcatctacaa ggtgaagctg cgcggcacca acttccctcc ggacggcccc gtaatgcaga
     2401 agaagaccat gggctgggag gctacgcgtg accaactgac tgaagagcag atcgcagaat
     2461 ttaaagaggc tttctcctgc tttgacaagg acggggatgg gacgataaca accaaggagc
     2521 ttgggacggt gatgcgatcg ctggggcaga accccacaga agcagagctg caggacatga
     2581 tcaatgaagt agatgccgac ggtgacggca cattcgactt ccctgagttc ctgacgatga
     2641 tggcaagaaa aatgaatgac acagacagtg aagaggaaat tcgcgaagcg ttccgcgtgt
     2701 ttgataagga cggcaatggc tacatcggcg cagcagagct tcgccacgtg atgacagacc
     2761 ttggagagaa gttaacagat gaggaggttg atgaaatgat cagggtagca gacatcgatg
     2821 gggatggtca ggtaaactac gaagagtttg tacaaatgat gacagcgaag tgaaagctct
     2881 agtataactt cgtatagcat acattatacg aagttatggg tcgatggtga tgcttggcaa
     2941 ttcgggtcga tggtgaagca ttggtcttcc tattttgata taacttcgta taggatactt
     3001 tatacgaagt tatggggatc cttgctagca gcttgaattc gactgcgctc tcaggcacga
     3061 cacgactcct ccgctgccca ccgcagactg aggcagcgct gagtcgccgg cgccgcagcg
     3121 cagatggtcg cgcccgtgcc cccctatctc gcgcctcgcg tggtgcggtc cggctgggcc
     3181 ggcggcggcg cggacgcgac caaggtggcc gggaagggga gtttgcgggg gaccggcgag
     3241 tgacgtcagc gcgccttcag tgctgaggcg gcggtggcgc gcgccgccag gcgggggcga
     3301 aggcactgtc cgcggtgctg aagctggcag tgcgcacgcg cctcgccgca tcctgtttcc
     3361 cctccccctc tctgataggg gatgcgcaat ttggggaatg ggggttgggt gcttgtccag
     3421 tgggtcgggg tcggtcgtca ggtaggcacc cccaccccgc ctcatcctgg tcctaaaacc
     3481 cacttgcact catacgcagg gccctctgca gagatcttcc tagagcatgg ctacgtagat
     3541 aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact
     3601 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     3661 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctggcgtaa tagcgaagag
     3721 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     3781 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3841 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3901 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3961 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4021 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4081 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4141 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4201 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     4261 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4321 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4381 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4441 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4501 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4561 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4621 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4681 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4741 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4801 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4861 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4921 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4981 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5041 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5101 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5161 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5221 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5281 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5341 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5401 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5461 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5521 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5581 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5641 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5701 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5761 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5821 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5881 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5941 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6001 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     6061 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     6121 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     6181 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     6241 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     6301 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     6361 acacaggaaa cagctatgac catgattacg ccaagctttg ct
//
