LOCUS       Exported                7005 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses the SthK channel.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-CamkII-Sthk-p2a-tpPAC minWPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7005)
  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 7005)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7005
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   47..187
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   58..187
                     /label=AAV2 ITR
     CDS             1226..1237
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             1939..1995
                     /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             3088..3117
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3127..3144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3160..3864
                     /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(3302..3320)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3570..3589
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3934..3954)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4314..4454
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4314..4443
                     /label=AAV2 ITR
     rep_origin      4529..4984
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4616..4635)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4826..4847
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5001..5020)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5120..5142
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5180..5198)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5266..5370
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5371..6231
                     /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(5589..5608)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6402..6990
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6891..6910
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg
       61 cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt
      121 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
      181 ggttcctgcg gccgcagatg ctgcacactt gtggactaag tttgttcgca tccccttctc
      241 caaccccctc agtacatcac cctgggggaa cagggtccac ttgctcctgg gcccacacag
      301 tcctgcagta ttgtgtatat aaggccaggg caaagaggag caggttttaa agtgaaaggc
      361 aggcaggtgt tggggaggca gttaccgggg caacgggaac agggcgtttc ggaggtggtt
      421 gccatgggga cctggatgct gacgaaggct cgcgaggctg tgagcagcca cagtgccctg
      481 ctcagaagcc ccaagctcgt cagtcaagcc ggttctccgt ttgcactcag gagcacgggc
      541 aggcgagtgg cccctagttc tgggggcagc gtgaaaagac ggtatcgata agcttgattc
      601 gagctagcca tgaaaagctc cgccttctcc caccccacct acaccctggt ctggaaagtc
      661 ggcattctgg ctgtcactct gtattacgct attcgaatcc cactgaccct ggtgttcccc
      721 tctctgttta gtcccctgct gcctctggat atcctggcca gtctggctct gatcgcagac
      781 attcctctgg atttcgcctt tgagtcacga aagacaagcg gcaggaaacc aactctgctg
      841 gctcctagcc gactgccaga tctgctggcc gctctgccac tggacctgct ggtgttcgcc
      901 ctgcacctgc catcacccct gagcctgctg tccctggtga ggctgctgaa gctgatctcc
      961 gtccagaggt ctgctacaag aatcctgtct tacagaatta acccagcact gctgcggctg
     1021 ctgagtctgg tgggattcat cctgctggca gcccatggga ttgcctgcgg ctggatgtca
     1081 ctgcagccac ctagcgagtc cccagcagga accagatacc tgagcgcctt ctactggaca
     1141 atcaccacac tgactaccat cggctacgga gatattaccc catccacacc cattcagacc
     1201 gtgtacacca tcgtcattga gctgctggga gctgcaatgt atggactggt catcgggaat
     1261 attgcatctc tggtcagtaa gctggacgcc gctaaactgc tgcaccgaga gaggatggaa
     1321 cgggtgacag ctttcctgag ttacaagaaa atctcacctg agctgcagag gagaattctg
     1381 gaatactttg attatctgtg ggagactcgg cgcgggtatg aggaacgcga ggtgctgaag
     1441 gaactgcctc acccactgcg actggctgtc gcaatggaaa tccatggcga cgtgattgag
     1501 aaggtcccac tgttcaaagg ggccggcgaa gactttatcc gcgatatcat tctgcatctg
     1561 gagcccgtga tctacggacc tggggaatat atcattaggg ctggcgagct gggcagcgat
     1621 gtctacttta tcaacagagg cagcgtggag gtcctgtccg cagacgaaaa gacccggtat
     1681 gccatcctgt ctgagggcca gttctttgga gaaatggcac tgattctgcg agcaccacga
     1741 acagctactg tgagagcacg gactttctgt gacctgtaca gactggataa agaaaccttt
     1801 gacagaatcc tgtctcgcta tcctgagatt gcagcccaga ttcaggaact ggctgtgcgg
     1861 aggaaagaag aactggaagg ggggacatca cggcggggaa ccggtcccgg gcttaaggag
     1921 ctcgcatgcg gaagcggagc tactaacttc agcctgctga agcaggctgg agacgtggag
     1981 gagaaccctg gacctagatc tggaattcaa ggcctctcga gcatgaaacg cctgacttac
     2041 attagcagct ttacacgccc actgatggct tctgaggtcg aggagattgg agcaaggtct
     2101 gtgaaaaata acacaagaga cggcctgact ggagtgctgt tctgctttaa caatatcttc
     2161 taccagattc tggagggacc cgaagatgcc ctgaaccgat gctatgctag aatcctgaag
     2221 gacaatcggc acaaagatat tttctgtctg gaggtcgaaa acaatatcac cgtgaggcag
     2281 tttggggact gggctatgaa gacagtgaga ctggacgaga gctccgattc tctgatccgg
     2341 cctattcgca caatgctgaa tagtctggca cgcactcatt acgtcctgga gcgatatagc
     2401 ccattcgaag tgctgcaggg actgcagaag ggagaggacc ccctggcctg ggtgctgaag
     2461 actagcgaaa aaaccatcct gttctccgac atcttcagca gtaccacatt cgcagagtcc
     2521 ctgtcaagca ccgagtttga acggctgctg ggcatctact atgatattgc caaccgctca
     2581 gtcatggaaa atggcggaac cgtgagcaag ctgacagggg acggcctgat ggcatacttt
     2641 aaagcctctg atacagtcca cgctctgagt gccgctatcg agatttgcag gagactggaa
     2701 gacgtgcgaa ctagtgcccc cgcttcagat cctgctcatt acctgtatgc aggagtcgga
     2761 ctgtgcagtg gacaggtgcg cgaggggaac atcggctcag cagccaagtt cgactataca
     2821 ctgctggggg attccgtgaa ctctgctgca agactggaag gcgtgtctcg gaaagtcaat
     2881 cctctggtca tctttgacga ttccctgctg aagcacatcg acaaaccatc tactctgaag
     2941 aaactgggcc tgtaccaggc caagggaaaa accgagaacc tgagcgtgta tactgtcgat
     3001 tacccctata cctccaggaa tatcaccatc ccagacctga aagcagccat tgccaagttt
     3061 aggacagcat ccagtgcagc aacgtacgag caaaagctga tctcggagga ggacctgaag
     3121 ggatcgcacc accatcacca ccatgcagct agatctgccg tgagcaaggg cgaggaggat
     3181 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     3241 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     3301 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
     3361 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
     3421 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     3481 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
     3541 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
     3601 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     3661 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
     3721 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
     3781 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
     3841 ggcggcatgg acgagctgta caagttaatt aactcgagat aatcaacctc tggattacaa
     3901 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3961 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     4021 cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg
     4081 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttta tttgtgaaat
     4141 ttgtgatgct attgctttat ttgtaaccat ctagctttat ttgtgaaatt tgtgatgcta
     4201 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     4261 attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcggc cgcaggaacc
     4321 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     4381 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     4441 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4501 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     4561 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4621 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4681 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4741 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4801 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4861 cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4921 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     4981 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     5041 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     5101 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     5161 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5221 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5281 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5341 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5401 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5461 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5521 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5581 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5641 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5701 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5761 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5821 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5881 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5941 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6001 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6061 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6121 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6181 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6241 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6301 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6361 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6421 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6481 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6541 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6601 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6661 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6721 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6781 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6841 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6901 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6961 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcg
//
