LOCUS       sequence-230192-        5510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EGFP and bReaChES under a minimal CamKII Promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5510)
  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 5510)
  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-CamKII-GFP-P2A-bReaches-minWPRE
FEATURES             Location/Qualifiers
     source          1..5510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      550..559
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(556..558,559..561,562..1272)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(556..558,559..561,562..1272)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(601..622)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(862..881)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1209..1230
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1288..1344
                     /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"
     primer_bind     complement(2393..2413)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   2773..2913
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2773..2902
                     /label=AAV2 ITR
     rep_origin      2988..3443
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3075..3094)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3285..3306
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3460..3479)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3579..3601
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3639..3657)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3725..3829
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3830..3898,3899..4690)
                     /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(4048..4067)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4861..5449
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5350..5369
                     /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 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      601 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      661 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
      721 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
      781 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
      841 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
      901 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
      961 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     1021 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     1081 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1141 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     1201 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1261 gagctgtaca aaccgcgggg atccggagct actaacttca gcctgctgaa gcaagcagga
     1321 gatgttgaag aaaaccccgg gcctatggac tatggcggcg ctttgtctgc cgtcggactg
     1381 ttccagacca gctacaccct ggagaacaac ggcagcgtga tctgcatccc caacaacggc
     1441 cagtgcttct gcctggcctg gctgaagagt aacggcacca acgccgagaa gctggccgcc
     1501 aacatcctgc agtgggtggt gtttgcgctg agcgtggcgt gcctgggctg gtatgcgtat
     1561 caggcgtggc gcgcgacctg cggctgggaa gaagtgtatg tggcgctgat tgaaatgatg
     1621 aaaagcatta ttgaagcgtt tcatgaattt gatagcccgg cgaccctgtg gctgagcagc
     1681 ggcaacggcg tggtgtggat gcgctatggc tcatggctgc tgacctgccc cgtgattctg
     1741 attcatctga gcaacctgac cggcctgaaa gatgattata gcaaacgcac catgggcctg
     1801 ctggtgagcg acgtgggctg cattgtgtgg ggcgcgacca gcgcgatgtg caccggctgg
     1861 accaaaattc tgttttttct gattagcctg agctatggca tgtataccta ttttcatgcg
     1921 gccaaagtgt atattgaagc gtttcatacc gtgccgaaag gcctgtgcag acagctggtg
     1981 agagccatgg cctggctgtt cttcgtgagc tggggcatgt tccccgtgct gttcctgctg
     2041 ggccccgagg gcttcggcca tattagcccg tatggcagcg cgattggcca tagcattctg
     2101 gatctgattg cgaagaacat gtggggcgtg ctgggcaact atctgcgcgt gaaaattcat
     2161 gaacatattc tgctgtatgg cgatattcgc aaaaaacaga aaattaccat tgcgggccag
     2221 gaaatggaag tggaaaccct ggtggcggaa gaagaagatg ggtcgaccaa gagcaggatc
     2281 accagcgagg gcgagtacat ccccctggac cagatcgaca tcaacgtgta actcgagata
     2341 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2401 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     2461 tggctttcat tttctcctcc ttgtataaat cctggttagt tcttgccacg gcggaactca
     2521 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     2581 tggtgtttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatc tagctttatt
     2641 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2701 aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt
     2761 taaagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     2821 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     2881 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     2941 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     3001 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3061 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3121 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3181 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     3241 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3301 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     3361 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3421 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     3481 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3541 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3601 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     3661 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     3721 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3781 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3841 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3901 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3961 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4021 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4081 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4141 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4201 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4261 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4321 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4381 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4441 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4501 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4561 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4621 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4681 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4741 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4801 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4861 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4921 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4981 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5041 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5101 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5161 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5221 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5281 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5341 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5401 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5461 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
