LOCUS       sequence-223382-        7407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-conditional expression of the SthK channel.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7407)
  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 7407)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2018-11-5"; volume: "9(1):4611. doi"; pages: " 
            10.1038/s41467-018-07038-8"
COMMENT     Sequence Label: pAAV-Syn-FLEX-Sthk-P2A-bPAC_minWPRE
FEATURES             Location/Qualifiers
     source          1..7407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        173..620
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(635..668)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(719..752)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(828..1535)
                     /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(1106..1125)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1375..1393
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(1551..1568)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1578..1607)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(2697..2753)
                     /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             complement(3455..3466)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    4095..4128
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    4179..4212
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(4274..4294)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4670..4810
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4670..4799
                     /label=AAV2 ITR
     rep_origin      4885..5340
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4972..4991)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5182..5203
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5357..5376)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5476..5498
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5536..5554)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5622..5726
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5727..5795,5796..6587)
                     /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(5945..5964)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6758..7346
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7247..7266
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtagagg gccctgcgta tgagtgcaag
      181 tgggttttag gaccaggatg aggcggggtg ggggtgccta cctgacgacc gaccccgacc
      241 cactggacaa gcacccaacc cccattcccc aaattgcgca tcccctatca gagaggggga
      301 ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc ttcagcaccg cggacagtgc
      361 cttcgccccc gcctggcggc gcgcgccacc gccgcctcag cactgaaggc gcgctgacgt
      421 cactcgccgg tcccccgcaa actccccttc ccggccacct tggtcgcgtc cgcgccgccg
      481 ccggcccagc cggaccgcac cacgcgaggc gcgagatagg ggggcacggg cgcgaccatc
      541 tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc ggaggagtcg
      601 tgtcgtgcct gagagcgcag tcgctagtac cggtataact tcgtatagga tactttatac
      661 gaagttatgc agaatggtag ctggattgta gctgctatta gcaatatgaa acctcttaat
      721 aacttcgtat agcatacatt atacgaagtt atttcgaact cgagccagag ctctggattt
      781 aaatggcgcg cctattaatt aaggccgcga ctctagaatt cgctagctta cttgtacagc
      841 tcgtccatgc cgccggtgga gtggcggccc tcggcgcgtt cgtactgttc cacgatggtg
      901 tagtcctcgt tgtgggaggt gatgtccaac ttgatgttga cgttgtaggc gccgggcagc
      961 tgcacgggct tcttggcctt gtaggtggtc ttgacctcag cgtcgtagtg gccgccgtcc
     1021 ttcagcttca gcctctgctt gatctcgccc ttcagggcgc cgtcctcggg gtacatccgc
     1081 tcggaggagg cctcccagcc catggtcttc ttctgcatta cggggccgtc ggaggggaag
     1141 ttggtgccgc gcagcttcac cttgtagatg aactcgccgt cctgcaggga ggagtcctgg
     1201 gtcacggtca ccacgccgcc gtcctcgaag ttcatcacgc gctcccactt gaagccctcg
     1261 gggaaggaca gcttcaagta gtcggggatg tcggcggggt gcttcacgta ggccttggag
     1321 ccgtacatga actgagggga caggatgtcc caggcgaagg gcagggggcc acccttggtc
     1381 accttcagct tggcggtctg ggtgccctcg taggggcggc cctcgccctc gccctcgatc
     1441 tcgaactcgt ggccgttcac ggagccctcc atgtgcacct tgaagcgcat gaactccttg
     1501 atgatggcca tgttatcctc ctcgcccttg ctcacggcag atctagctgc atggtggtga
     1561 tggtggtgcg atcccttcag gtcctcctcc gagatcagct tttgctcgta cgtccgcggc
     1621 ttgtcgtttt ccagggtctg ccggatcagc tgggtgatct gcagggtgtc gtagtacttc
     1681 tgggcctcat tcacggtgta gacctcgatg gcttcctgct tgcccttgac ctggtgggct
     1741 cccaggttga tgaaggtcca ctgggcctgg cagcacttct tcacgccggc tgtaaaggcc
     1801 agggcgtagg gcagctgtct tgtcagggct tccagcctgg cggccacgtt cacggcgtcc
     1861 cccagcaggg tgtggtccat cttcaggctg ctgcccatgt tgccctcgat cacgtggccg
     1921 tagctcaggc cgatgccggt gtacagcagg tgcagggggt tggtggcctc cacgtggtgc
     1981 cgcagctgct tcagctcgga gatgatgtcc aggctggtcc ggatggcggc gtcgccctgc
     2041 tctttggtga agctggccat cacgcagtcg ccgatgaact tggtcacttc gccgccgtag
     2101 gcgctgatga tccgggtgca gatgctgaag taccggttga ccaggatgac cacctcgttc
     2161 acgggcagct tctcggtcag ggtgctgaag gccaggatgt cgctgaagaa gatgatcttc
     2221 tcgaccagct ggggctccac ggtcaggggg ttgatgccct ggttgatcag gtagatcact
     2281 ctggcgggca tgtacttttc cagcacccgg tggctctggg tgatggtctg cagcaggctc
     2341 ttgatgggct ggatcatcag ctcgctgttc tcgttcaggt tgatggtttt catggcccag
     2401 ttggggaaca tccggtcggt gatgtcgtac tcggttttca ggcacaggat gttggtgtgc
     2461 cggtcgtcca ccaggatttt cttgtacagc ttgtccacct tctcgttctc gccttccagg
     2521 atctggaaga acaggccctg caggtacagc agcacgccgg tgatgttgtc ccgctggttg
     2581 ttcttgatgg acaccttgcc gatccgctgg atctcttcca ggctcaggtg gccgctgatc
     2641 ttgctgatgt acaccagccg cttcatcatg ctcgagaggc cttgaattcc agatctaggt
     2701 ccagggttct cctccacgtc tccagcctgc ttcagcaggc tgaagttagt agctccgctt
     2761 ccgcatgcga gctccttaag cccgggaccg gttccccgcc gtgatgtccc cccttccagt
     2821 tcttctttcc tccgcacagc cagttcctga atctgggctg caatctcagg atagcgagac
     2881 aggattctgt caaaggtttc tttatccagt ctgtacaggt cacagaaagt ccgtgctctc
     2941 acagtagctg ttcgtggtgc tcgcagaatc agtgccattt ctccaaagaa ctggccctca
     3001 gacaggatgg cataccgggt cttttcgtct gcggacagga cctccacgct gcctctgttg
     3061 ataaagtaga catcgctgcc cagctcgcca gccctaatga tatattcccc aggtccgtag
     3121 atcacgggct ccagatgcag aatgatatcg cggataaagt cttcgccggc ccctttgaac
     3181 agtgggacct tctcaatcac gtcgccatgg atttccattg cgacagccag tcgcagtggg
     3241 tgaggcagtt ccttcagcac ctcgcgttcc tcatacccgc gccgagtctc ccacagataa
     3301 tcaaagtatt ccagaattct cctctgcagc tcaggtgaga ttttcttgta actcaggaaa
     3361 gctgtcaccc gttccatcct ctctcggtgc agcagtttag cggcgtccag cttactgacc
     3421 agagatgcaa tattcccgat gaccagtcca tacattgcag ctcccagcag ctcaatgacg
     3481 atggtgtaca cggtctgaat gggtgtggat ggggtaatat ctccgtagcc gatggtagtc
     3541 agtgtggtga ttgtccagta gaaggcgctc aggtatctgg ttcctgctgg ggactcgcta
     3601 ggtggctgca gtgacatcca gccgcaggca atcccatggg ctgccagcag gatgaatccc
     3661 accagactca gcagccgcag cagtgctggg ttaattctgt aagacaggat tcttgtagca
     3721 gacctctgga cggagatcag cttcagcagc ctcaccaggg acagcaggct caggggtgat
     3781 ggcaggtgca gggcgaacac cagcaggtcc agtggcagag cggccagcag atctggcagt
     3841 cggctaggag ccagcagagt tggtttcctg ccgcttgtct ttcgtgactc aaaggcgaaa
     3901 tccagaggaa tgtctgcgat cagagccaga ctggccagga tatccagagg cagcagggga
     3961 ctaaacagag aggggaacac cagggtcagt gggattcgaa tagcgtaata cagagtgaca
     4021 gccagaatgc cgactttcca gaccagggtg taggtggggt gggagaaggc ggagcttttc
     4081 atggtggcgg atccataact tcgtataaag tatcctatac gaagttattt gccttaaccc
     4141 agaaattatc actgttattc tttagaatgg tgcaaagaat aacttcgtat aatgtatgct
     4201 atacgaagtt atgtcgagat aatcaacctc tggattacaa aatttgtgaa agattgactg
     4261 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     4321 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttag
     4381 ttcttgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     4441 tgttgggcac tgacaattcc gtggtgttta tttgtgaaat ttgtgatgct attgctttat
     4501 ttgtaaccat tctagcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     4561 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     4621 gggggagatg tgggaggttt tttaaagcac gtgcggaccg agcggccgca ggaaccccta
     4681 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     4741 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     4801 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4861 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4921 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4981 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5041 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5101 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5161 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     5221 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     5281 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     5341 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     5401 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     5461 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     5521 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     5581 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5641 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5701 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     5761 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5821 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     5881 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     5941 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     6001 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     6061 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6121 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     6181 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6241 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     6301 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     6361 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6421 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6481 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6541 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6601 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6661 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6721 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6781 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6841 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6901 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6961 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     7021 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     7081 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     7141 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     7201 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     7261 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     7321 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     7381 ccttttgctg gccttttgct cacatgt
//
