LOCUS       sequence-379291-        7537 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-vector for bidirectional optogenetic manipulation of 
            tetTag-positive neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7537)
  AUTHORS   Wiegert Lab
  TITLE     Wiegert Lab Plasmids 2022
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7537)
  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: 
            "Unpublished"
COMMENT     Sequence Label: TRE3GV-somBiPOLES-mCerulean
FEATURES             Location/Qualifiers
     source          1..7537
                     /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        2680..3029
                     /label=TRE3GV promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, optimized for 
                     retroviral and lentiviral vectors"
     protein_bind    2688..2706
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2724..2742
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2760..2778
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2796..2814
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2832..2850
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2868..2886
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2904..2922
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2968..2992
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(4052..4073)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4313..4332)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4660..4681
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5081..6124
                     /codon_start=1
                     /product="red light-drivable channelrhodopsin from 
                     Chlamydomonas noctigama (Klapoetke et al., 2014)"
                     /label=Chrimson
                     /translation="ELISSATRSLFAAGGINPWPNPYHHEDMGCGGMTPTGECFSTEWW
                     CDPSYGLSDAGYGYCFVEATGGYLVVGVEKKQAWLHSRGTPGEKIGAQVCQWIAFSIAI
                     ALLTFYGFSAWKATCGWEEVYVCCVEVLFVTLEIFKEFSSPATVYLSTGNHAYCLRYFE
                     WLLSCPVILIKLSNLSGLKNDYSKRTMGLIVSCVGMIVFGMAAGLATDWLKWLLYIVSC
                     IYGGYMYFQAAKCYVEANHSVPKGHCRMVVKLMAYAYFASWGSYPILWAVGPEGLLKLS
                     PYANSIGHSICDIIAKEFWTFLAHHLRIKIHEHILIHGDIRKTTKMEIGGEEVEVEEFV
                     EEEDEDTV"
     misc_feature    6347..6935
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6400..6420)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6818..6829)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6976..7183
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7353..7482
                     /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 ggttcgatga ggccctttcg tcttcactcg agtttactcc ctatcagtga
     2701 tagagaacgt atgaagagtt tactccctat cagtgataga gaacgtatgc agactttact
     2761 ccctatcagt gatagagaac gtataaggag tttactccct atcagtgata gagaacgtat
     2821 gaccagttta ctccctatca gtgatagaga acgtatctac agtttactcc ctatcagtga
     2881 tagagaacgt atatccagtt tactccctat cagtgataga gaacgtatgt cgaggtaggc
     2941 gtgtacggtg ggcgcctata aaagcagagc tcgtttagtg aaccgtcaga tcgcctggag
     3001 caattccaca acacttttgt cttatacttg gatccgggcc cgcggccgat ccatggcatc
     3061 acaggtcgtc tatggggaat gggcatcaac tcacaccgag tgctacaata tgtccagaat
     3121 cgactccacc ttcgtctcac tgctgcagct ggtgtgggcc gtggtctcag gctgccagac
     3181 aatcttcatg attagcaggg ctcccaaggt cccttgggag tccgtgtacc tgccctttgt
     3241 cgaatctatc acttatgcac tggccagtac tggcaacgga accctgcaga tgagggacgg
     3301 cagattcttt ccttggagcc gaatggcctc ctggctgtgc acctgtccaa tcatgctggg
     3361 acagatttct aacatggctc tggtgaagta caaaagtatc ccactgaatc ccattgcaca
     3421 ggccgcttcc atcattagag tggtcatggg aatcaccgct acaatttctc ccgcagagta
     3481 tatgaagtgg ctgttctttt tctttggggc cacctgcctg gtgttcgagt acagtgtggt
     3541 cttcacaatc tttcaagtgg ggctgtatgg ctttgaatct gtcgggacac ctctggcaca
     3601 gaaggtggtg gtgaggatca agatgctgcg cctgatcttc tttattgctt ggactatgtt
     3661 cccaatcgtg tggctgattt cacccaccgg agtctgcgtg atccacgaga acgtgagcgc
     3721 cattctgtac ctgctggctg acggcctgtg caaaaataca tatggagtga tcctgtggag
     3781 cactgcctgg ggagtgctgg aagggaagtg ggaccccgca tgtctgccag ggcaggagaa
     3841 acccgaagcc gacgatcctt ttggcctgaa tcacgagaag aatgcccctc ctaacgacga
     3901 agtcaatatc agaatgtttg gaaggggacg tacggcggcc aagagcagga tcaccagcga
     3961 gggcgagtac atccccctgg accagatcga catcaacgtg gcggtacccg tgagcaaggg
     4021 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     4081 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     4141 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     4201 gagctggggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     4261 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     4321 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     4381 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     4441 cgccatccac ggcaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa
     4501 cttcggcctc aactgcaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     4561 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     4621 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     4681 gaccgccgcc gggatcactc tcggcatgga cgagctgtat accggtgcag ccctgcagga
     4741 gaagaagtca tgcagccagc gcatggccga attccggcaa tactgttgga acccggacac
     4801 tgggcagatg ctgggccgca ccccagcccg gtgggtgtgg atcagcctgt actatgcagc
     4861 tttctacgtg gtcatgactg ggctctttgc cttgtgcatc tatgtgctga tgcagaccat
     4921 tgatccctac acccccgact accaggacca gttaaagtca ccgggggtaa ccttgagacc
     4981 ggatgtgtat ggggaaagag ggctgcagat ttcctacaac atctctgaaa acagctctag
     5041 acaggcccag atcaccggac gtccggaggg agctgtacaa gagctgatca gcagcgccac
     5101 cagatctctg tttgccgccg gaggcatcaa cccttggcct aacccctacc accacgagga
     5161 catgggctgt ggaggaatga cacctacagg cgagtgcttc agcaccgagt ggtggtgtga
     5221 cccttcttac ggactgagcg acgccggata cggatattgc ttcgtggagg ccacaggcgg
     5281 ctacctggtc gtgggagtgg agaagaagca ggcttggctg cacagcagag gcacaccagg
     5341 agaaaagatc ggcgcccagg tctgccagtg gattgctttc agcatcgcca tcgccctgct
     5401 gacattctac ggcttcagcg cttggaaggc cacttgcggt tgggaggagg tctacgtctg
     5461 ttgcgtcgag gtgctgttcg tgaccctgga gatcttcaag gagttcagca gccccgccac
     5521 agtgtacctg tctaccggca accacgccta ttgcctgcgc tacttcgagt ggctgctgtc
     5581 ttgccccgtg atcctgatca agctgagcaa cctgagcggc ctgaagaacg actacagcaa
     5641 gcggaccatg ggcctgatcg tgtcttgcgt gggaatgatc gtgttcggca tggccgcagg
     5701 actggctacc gattggctca agtggctgct gtatatcgtg tcttgcatct acggcggcta
     5761 catgtacttc caggccgcca agtgctacgt ggaagccaac cacagcgtgc ctaaaggcca
     5821 ttgccgcatg gtcgtgaagc tgatggccta cgcttacttc gcctcttggg gcagctaccc
     5881 aatcctctgg gcagtgggac cagaaggact gctgaagctg agcccttacg ccaacagcat
     5941 cggccacagc atctgcgaca tcatcgccaa ggagttttgg accttcctgg cccaccacct
     6001 gaggatcaag atccacgagc acatcctgat ccacggcgac atccggaaga ccaccaagat
     6061 ggagatcgga ggcgaggagg tggaagtgga agagttcgtg gaggaggagg acgaggacac
     6121 agtgctcgag cagagccagc ccatcctgaa caccaaggag atggcccccc agagcaagcc
     6181 ccccgaggag ctggagatga gcagcatgcc cagccccgtg gcccccctgc ccgcccgcac
     6241 cgagggcgtg atcgacatgc gcagcatgag cagcatcgac agcttcatca gctgcgccac
     6301 cgacttcccc gaggccaccc gcttctgatc tagaaagctt atcgataatc aacctctgga
     6361 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     6421 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     6481 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     6541 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     6601 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     6661 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     6721 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     6781 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     6841 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     6901 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac ccgggcggcc
     6961 gctcataaag ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     7021 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     7081 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     7141 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtc cggactgtac
     7201 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag
     7261 taaaacctct acaaatgtgg taaaatcgat aaggatcttc ctagagcatg gctacgtaga
     7321 taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac
     7381 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     7441 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcatta atgaatcggc
     7501 caacgcgcgg ggagaggcgg tttgcgtatt gggcgct
//
