LOCUS       Exported                5580 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses bPAC(WT)-mCherry under a minimal CamKII Promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-CamkII-bPac(WT)-mCherry-minWPRE.sbd
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5580)
  AUTHORS   Bernal Sierra YA, Rost BR, Pofahl M, Fernandes AM, Kopton RA, Moser 
            S, Holtkamp D, Masala N, Beed P, Tukker JJ, Oldani S, Bonigk W, Kohl
            P, Baier H, Schneider-Warme F, Hegemann P, Beck H, Seifert R, 
            Schmitz D
  TITLE     Potassium channel-based optogenetic silencing.
  JOURNAL   Nat Commun. 2018 Nov 5;9(1):4611. doi: 10.1038/s41467-018-07038-8.
  PUBMED    30397200
REFERENCE   2  (bases 1 to 5580)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5580
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             1614..1643
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1653..1670
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1686..2393
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1828..1846)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2096..2115
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2463..2483)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   2843..2983
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2843..2972
                     /label=AAV2 ITR
     rep_origin      3058..3513
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3145..3164)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3355..3376
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3530..3549)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3649..3671
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3709..3727)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3795..3899
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3900..4760
                     /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(4118..4137)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4931..5519
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5420..5439
                     /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 gcggccgcag atgctgcaca cttgtggact aagtttgttc gcatcccctt
      181 ctccaacccc ctcagtacat caccctgggg gaacagggtc cacttgctcc tgggcccaca
      241 cagtcctgca gtattgtgta tataaggcca gggcaaagag gagcaggttt taaagtgaaa
      301 ggcaggcagg tgttggggag gcagttaccg gggcaacggg aacagggcgt ttcggaggtg
      361 gttgccatgg ggacctggat gctgacgaag gctcgcgagg ctgtgagcag ccacagtgcc
      421 ctgctcagaa gccccaagct cgtcagtcaa gccggttctc cgtttgcact caggagcacg
      481 ggcaggcgag tggcccctag ttctgggggc agcgtgaaaa gacggtatcg ataagcttga
      541 ttcgagctag catgatgaag cggctggtgt acatcagcaa gatcagcggc cacctgagcc
      601 tggaagagat ccagcggatc ggcaaggtgt ccatcaagaa caaccagcgg gacaacatca
      661 ccggcgtgct gctgtacctg cagggcctgt tcttccagat cctggaaggc gagaacgaga
      721 aggtggacaa gctgtacaag aaaatcctgg tggacgaccg gcacaccaac atcctgtgcc
      781 tgaaaaccga gtacgacatc accgaccgga tgttccccaa ctgggccatg aaaaccatca
      841 acctgaacga gaacagcgag ctgatgatcc agcccatcaa gagcctgctg cagaccatca
      901 cccagagcca ccgggtgctg gaaaagtaca tgcccgccag agtgatctac ctgatcaacc
      961 agggcatcaa ccccctgacc gtggagcccc agctggtcga gaagatcatc ttcttcagcg
     1021 acatcctggc cttcagcacc ctgaccgaga agctgcccgt gaacgaggtg gtcatcctgg
     1081 tcaaccggta cttcagcatc tgcacccgga tcatcagcgc ctacggcggc gaagtgacca
     1141 agttcatcgg cgactgcgtg atggccagct tcaccaaaga gcagggcgac gccgccatcc
     1201 ggaccagcct ggacatcatc tccgagctga agcagctgcg gcaccacgtg gaggccacca
     1261 accccctgca cctgctgtac accggcatcg gcctgagcta cggccacgtg atcgagggca
     1321 acatgggcag cagcctgaag atggaccaca ccctgctggg ggacgccgtg aacgtggccg
     1381 ccaggctgga agccctgaca agacagctgc cctacgccct ggcctttaca gccggcgtga
     1441 agaagtgctg ccaggcccag tggaccttca tcaacctggg agcccaccag gtcaagggca
     1501 agcaggaagc catcgaggtc tacaccgtga atgaggccca gaagtactac gacaccctgc
     1561 agatcaccca gctgatccgg cagaccctgg aaaacgacaa gccgcggacg tacgagcaaa
     1621 agctgatctc ggaggaggac ctgaagggat cgcaccacca tcaccaccat gcagctagat
     1681 ctgccgtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     1741 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     1801 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     1861 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     1921 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     1981 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     2041 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     2101 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     2161 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     2221 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     2281 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     2341 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag taattaatta
     2401 actcgagata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2461 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2521 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttagt tcttgccacg
     2581 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2641 gacaattccg tggtgtttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatc
     2701 tagctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2761 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     2821 ggaggttttt taaagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     2881 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     2941 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     3001 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     3061 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3121 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3181 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3241 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     3301 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3361 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     3421 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3481 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     3541 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     3601 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     3661 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     3721 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     3781 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3841 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3901 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3961 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4021 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4081 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4141 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4201 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4261 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4321 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4381 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4441 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4501 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4561 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4621 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4681 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4741 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4801 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4861 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4921 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4981 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5041 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5101 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5161 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5221 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5281 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5341 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5401 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5461 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5521 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
