LOCUS       sequence-389374-        7569 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-vector for bidirectional optogenetic manipulation of neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7569)
  AUTHORS   Vierock J, Rodriguez-Rozada S, Dieter A, Pieper F, Sims R, Tenedini 
            F, Bergs ACF, Bendifallah I, Zhou F, Zeitzschel N, Ahlbeck J, 
            Augustin S, Sauter K, Papagiakoumou E, Gottschalk A, Soba P, 
            Emiliani V, Engel AK, Hegemann P, Wiegert JS
  TITLE     BiPOLES is an optogenetic tool developed for bidirectional 
            dual-color control of neurons.
  JOURNAL   Nat Commun. 2021 Jul 26;12(1):4527. doi: 10.1038/s41467-021-24759-5.
  PUBMED    34312384
REFERENCE   2  (bases 1 to 7569)
  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: "2021-07-26"; volume: "12(1):4527. doi"; pages: " 
            10.1038/s41467-021-24759-5"
COMMENT     Sequence Label: hSyn-GtACR2-ChRmine-tandem-eYFP
FEATURES             Location/Qualifiers
     source          1..7569
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2069..2090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2281..2300
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2459..2588
                     /label=AAV2 ITR
     promoter        2688..3135
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             join(5449..5451,5452..6162)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5491..5512)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5752..5771)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6099..6120
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6379..6967
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6432..6452)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6850..6861)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    7008..7215
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7385..7514
                     /label=AAV2 ITR
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
     2641 gaagatcgta ggttaattaa tctagactgc agagggccct gcgtatgagt gcaagtgggt
     2701 tttaggacca ggatgaggcg gggtgggggt gcctacctga cgaccgaccc cgacccactg
     2761 gacaagcacc caacccccat tccccaaatt gcgcatcccc tatcagagag ggggagggga
     2821 aacaggatgc ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac agtgccttcg
     2881 cccccgcctg gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc
     2941 gccggtcccc cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc cgccgccggc
     3001 ccagccggac cgcaccacgc gaggcgcgag ataggggggc acgggcgcga ccatctgcgc
     3061 tgcggcgccg gcgactcagc gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg
     3121 tgcctgagag cgcagtcgag aggatccatg gcatcacagg tcgtctatgg ggaatgggca
     3181 tcaactcaca ccgagtgcta caatatgtcc agaatcgact ccaccttcgt ctcactgctg
     3241 cagctggtgt gggccgtggt ctcaggctgc cagacaatct tcatgattag cagggctccc
     3301 aaggtccctt gggagtccgt gtacctgccc tttgtcgaat ctatcactta tgcactggcc
     3361 agtactggca acggaaccct gcagatgagg gacggcagat tctttccttg gagccgaatg
     3421 gcctcctggc tgtgcacctg tccaatcatg ctgggacaga tttctaacat ggctctggtg
     3481 aagtacaaaa gtatcccact gaatcccatt gcacaggccg cttccatcat tagagtggtc
     3541 atgggaatca ccgctacaat ttctcccgca gagtatatga agtggctgtt ctttttcttt
     3601 ggggccacct gcctggtgtt cgagtacagt gtggtcttca caatctttca agtggggctg
     3661 tatggctttg aatctgtcgg gacacctctg gcacagaagg tggtggtgag gatcaagatg
     3721 ctgcgcctga tcttctttat tgcttggact atgttcccaa tcgtgtggct gatttcaccc
     3781 accggagtct gcgtgatcca cgagaacgtg agcgccattc tgtacctgct ggctgacggc
     3841 ctgtgcaaaa atacatatgg agtgatcctg tggagcactg cctggggagt gctggaaggg
     3901 aagtgggacc ccgcatgtct gccagggcag gagaaacccg aagccgacga tccttttggc
     3961 ctgaatcacg agaagaatgc ccctcctaac gacgaagtca atatcagaat gtttggaagg
     4021 ggacgtacgg cggccaagag caggatcacc agcgagggcg agtacatccc cctggaccag
     4081 atcgacatca acgtggcggt agcagccctg caggagaaga agtcatgcag ccagcgcatg
     4141 gccgaattcc ggcaatactg ttggaacccg gacactgggc agatgctggg ccgcacccca
     4201 gcccggtggg tgtggatcag cctgtactat gcagctttct acgtggtcat gactgggctc
     4261 tttgccttgt gcatctatgt gctgatgcag accattgatc cctacacccc cgactaccag
     4321 gaccagttaa agtcaccggg ggtaaccttg agaccggatg tgtatgggga aagagggctg
     4381 cagatttcct acaacatctc tgaaaacagc tctagacagg cccagatcac cggacgtccg
     4441 gagggagctg taatggcaca cgcaccaggc accgaccaga tgttctacgt gggcacaatg
     4501 gacggctggt atctggatac caagctgaac tccgtggcca tcggcgccca ctggtcttgc
     4561 tttatcgtgc tgacaatcac cacattctac ctgggctatg agtcctggac atctcggggc
     4621 cctagcaaga gaacctcctt ttacgccggc tatcaggagg agcagaacct ggccctgttt
     4681 gtgaatttct ttgccatgct gagctacttc ggcaagatcg tggccgacac actgggccac
     4741 aacttcggcg atgtgggccc cttcatcatc ggcttcggca attacaggta tgccgactac
     4801 atgctgacct gcccaatgct ggtgtacgat ctgctgtatc agctgagggc cccctatcgc
     4861 gtgtcttgta gcgccatcat ctttgccatc ctgatgtctg gcgtgctggc cgagttctac
     4921 gcagagggcg accctaggct gaggaatggc gcctacgcct ggtatggctt tggctgtttc
     4981 tggtttatct tcgcctactc tatcgtgatg agcatcgtgg ccaagcagta tagccggctg
     5041 gcccagctgg cccaggatac aggcgccgag cacagcctgc acgtgctgaa gttcgccgtg
     5101 tttaccttct ccatgctgtg gattctgttt cccctggtgt gggccatctg ccctagaggc
     5161 ttcggctgga tcgacgataa ctggacagag gtggcccact gcgtgtgcga catcgtggcc
     5221 aagtcctgtt acggctttgc cctggcccgg ttcagaaaga cctatgatga ggagctgttt
     5281 cggctgctgg agcagctggg acacgacgag gatgagttcc agaagctgga gctggatatg
     5341 aggctgagca gcaatggcga gcgcctgcgg agactgtctg cggccgccaa gagcaggatc
     5401 accagcgagg gcgagtacat ccccctggac cagatcgaca tcaacgtggt gagcaagggc
     5461 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     5521 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     5581 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc
     5641 ggctacggcc tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     5701 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     5761 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     5821 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     5881 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     5941 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     6001 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     6061 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     6121 accgccgccg ggatcactct cggcatggac gagctgtaca agcagagcca gcccatcctg
     6181 aacaccaagg agatggcccc ccagagcaag ccccccgagg agctggagat gagcagcatg
     6241 cccagccccg tggcccccct gcccgcccgc accgagggcg tgatcgacat gcgcagcatg
     6301 agcagcatcg acagcttcat cagctgcgcc accgacttcc ccgaggccac ccgcttctga
     6361 tctagaaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     6421 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     6481 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     6541 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     6601 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     6661 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     6721 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     6781 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     6841 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     6901 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     6961 tccccgcatc gataccgtcg acccgggcgg ccgctcataa agctcgactg tgccttctag
     7021 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     7081 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     7141 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     7201 caggcatgct ggggatgcgg tccggactgt acaattgcat tcattttatg tttcaggttc
     7261 agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg
     7321 ataaggatct tcctagagca tggctacgta gataagtagc atggcgggtt aatcattaac
     7381 tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     7441 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     7501 gagcgagcgc gcagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     7561 ttkggcgct
//
