LOCUS       Exported                6991 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses the SthK channel.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-camk2-Sthk-p2a-bpac(WT) minWPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6991)
  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 6991)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6991
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   47..176
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             1215..1226
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             1928..1984
                     /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             3074..3103
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3113..3130
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3146..3850
                     /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(3288..3306)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3556..3575
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3920..3940)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4300..4440
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4300..4429
                     /label=AAV2 ITR
     rep_origin      4515..4970
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4602..4621)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4812..4833
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4987..5006)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5106..5128
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5166..5184)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5252..5356
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5357..6217
                     /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(5575..5594)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6388..6976
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6877..6896
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg
       61 cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc
      121 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg
      181 ccgcagatgc tgcacacttg tggactaagt ttgttcgcat ccccttctcc aaccccctca
      241 gtacatcacc ctgggggaac agggtccact tgctcctggg cccacacagt cctgcagtat
      301 tgtgtatata aggccagggc aaagaggagc aggttttaaa gtgaaaggca ggcaggtgtt
      361 ggggaggcag ttaccggggc aacgggaaca gggcgtttcg gaggtggttg ccatggggac
      421 ctggatgctg acgaaggctc gcgaggctgt gagcagccac agtgccctgc tcagaagccc
      481 caagctcgtc agtcaagccg gttctccgtt tgcactcagg agcacgggca ggcgagtggc
      541 ccctagttct gggggcagcg tgaaaagacg gtatcgataa gcttgattcg agctagccat
      601 gaaaagctcc gccttctccc accccaccta caccctggtc tggaaagtcg gcattctggc
      661 tgtcactctg tattacgcta ttcgaatccc actgaccctg gtgttcccct ctctgtttag
      721 tcccctgctg cctctggata tcctggccag tctggctctg atcgcagaca ttcctctgga
      781 tttcgccttt gagtcacgaa agacaagcgg caggaaacca actctgctgg ctcctagccg
      841 actgccagat ctgctggccg ctctgccact ggacctgctg gtgttcgccc tgcacctgcc
      901 atcacccctg agcctgctgt ccctggtgag gctgctgaag ctgatctccg tccagaggtc
      961 tgctacaaga atcctgtctt acagaattaa cccagcactg ctgcggctgc tgagtctggt
     1021 gggattcatc ctgctggcag cccatgggat tgcctgcggc tggatgtcac tgcagccacc
     1081 tagcgagtcc ccagcaggaa ccagatacct gagcgccttc tactggacaa tcaccacact
     1141 gactaccatc ggctacggag atattacccc atccacaccc attcagaccg tgtacaccat
     1201 cgtcattgag ctgctgggag ctgcaatgta tggactggtc atcgggaata ttgcatctct
     1261 ggtcagtaag ctggacgccg ctaaactgct gcaccgagag aggatggaac gggtgacagc
     1321 tttcctgagt tacaagaaaa tctcacctga gctgcagagg agaattctgg aatactttga
     1381 ttatctgtgg gagactcggc gcgggtatga ggaacgcgag gtgctgaagg aactgcctca
     1441 cccactgcga ctggctgtcg caatggaaat ccatggcgac gtgattgaga aggtcccact
     1501 gttcaaaggg gccggcgaag actttatccg cgatatcatt ctgcatctgg agcccgtgat
     1561 ctacggacct ggggaatata tcattagggc tggcgagctg ggcagcgatg tctactttat
     1621 caacagaggc agcgtggagg tcctgtccgc agacgaaaag acccggtatg ccatcctgtc
     1681 tgagggccag ttctttggag aaatggcact gattctgcga gcaccacgaa cagctactgt
     1741 gagagcacgg actttctgtg acctgtacag actggataaa gaaacctttg acagaatcct
     1801 gtctcgctat cctgagattg cagcccagat tcaggaactg gctgtgcgga ggaaagaaga
     1861 actggaaggg gggacatcac ggcggggaac cggtcccggg cttaaggagc tcgcatgcgg
     1921 aagcggagct actaacttca gcctgctgaa gcaggctgga gacgtggagg agaaccctgg
     1981 acctagatct ggaattcaag gcctctcgag catgatgaag cggctggtgt acatcagcaa
     2041 gatcagcggc cacctgagcc tggaagagat ccagcggatc ggcaaggtgt ccatcaagaa
     2101 caaccagcgg gacaacatca ccggcgtgct gctgtacctg cagggcctgt tcttccagat
     2161 cctggaaggc gagaacgaga aggtggacaa gctgtacaag aaaatcctgg tggacgaccg
     2221 gcacaccaac atcctgtgcc tgaaaaccga gtacgacatc accgaccgga tgttccccaa
     2281 ctgggccatg aaaaccatca acctgaacga gaacagcgag ctgatgatcc agcccatcaa
     2341 gagcctgctg cagaccatca cccagagcca ccgggtgctg gaaaagtaca tgcccgccag
     2401 agtgatctac ctgatcaacc agggcatcaa ccccctgacc gtggagcccc agctggtcga
     2461 gaagatcatc ttcttcagcg acatcctggc cttcagcacc ctgaccgaga agctgcccgt
     2521 gaacgaggtg gtcatcctgg tcaaccggta cttcagcatc tgcacccgga tcatcagcgc
     2581 ctacggcggc gaagtgacca agttcatcgg cgactgcgtg atggccagct tcaccaaaga
     2641 gcagggcgac gccgccatcc ggaccagcct ggacatcatc tccgagctga agcagctgcg
     2701 gcaccacgtg gaggccacca accccctgca cctgctgtac accggcatcg gcctgagcta
     2761 cggccacgtg atcgagggca acatgggcag cagcctgaag atggaccaca ccctgctggg
     2821 ggacgccgtg aacgtggccg ccaggctgga agccctgaca agacagctgc cctacgccct
     2881 ggcctttaca gccggcgtga agaagtgctg ccaggcccag tggaccttca tcaacctggg
     2941 agcccaccag gtcaagggca agcaggaagc catcgaggtc tacaccgtga atgaggccca
     3001 gaagtactac gacaccctgc agatcaccca gctgatccgg cagaccctgg aaaacgacaa
     3061 gccgcggacg tacgagcaaa agctgatctc ggaggaggac ctgaagggat cgcaccacca
     3121 tcaccaccat gcagctagat ctgccgtgag caagggcgag gaggataaca tggccatcat
     3181 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
     3241 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     3301 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     3361 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     3421 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     3481 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     3541 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
     3601 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
     3661 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
     3721 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
     3781 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     3841 gctgtacaag ttaattaact cgagataatc aacctctgga ttacaaaatt tgtgaaagat
     3901 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3961 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     4021 ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     4081 ctcggctgtt gggcactgac aattccgtgg tgtttatttg tgaaatttgt gatgctattg
     4141 ctttatttgt aaccatctag ctttatttgt gaaatttgtg atgctattgc tttatttgta
     4201 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     4261 gttcaggggg agatgtggga ggttttttaa agcggccgca ggaaccccta gtgatggagt
     4321 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     4381 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     4441 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     4501 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4561 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     4621 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4681 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     4741 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4801 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     4861 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     4921 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     4981 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     5041 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     5101 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     5161 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     5221 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     5281 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5341 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5401 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5461 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5521 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5581 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5641 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5701 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5761 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5821 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5881 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5941 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     6001 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6061 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     6121 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6181 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     6241 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6301 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6361 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6421 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6481 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6541 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6601 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6661 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6721 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6781 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6841 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6901 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6961 gcggagccta tggaaaaacg ccagcaacgc g
//
