LOCUS       sequence-379302-        5953 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AAV-vector for optogenetic silencing of noradrenergic neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5953)
  TITLE     PRSx8-Plasmids
REFERENCE   2  (bases 1 to 5953)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: PRS_8-somGtACR2-eGFP
FEATURES             Location/Qualifiers
     source          1..5953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1360..1362,1363..2073)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(1360..1362,1363..2073)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1402..1423)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1663..1682)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2010..2031
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2296..2884
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2349..2369)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2767..2778)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3053..3134
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3090..3109)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3144..3163
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3213..3353
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3213..3342
                     /label=AAV2 ITR
     rep_origin      3428..3883
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3515..3534)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3725..3746
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3900..3919)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4019..4041
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4079..4097)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4165..4269
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4270..4338,4339..5130)
                     /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(4488..4507)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5301..5889
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5790..5809
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(5951..5953,1..127)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt
       61 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
      121 ggttcctgcg gccgcacgcg tttaattaaa aacgcgtatt agcttccgct agacaaatgt
      181 gattaccccc gctagacaaa tgtgattacc cgcgctagac aaatgtgatt accccgctag
      241 acaaatgtga ttaccccccg ctagacaaat gtgattaccc ccgctagaca aatgtgatta
      301 cccgcgctag acaaatgtga ttaccccgct agacaaatgt gattaccccc gaccagggca
      361 taaatggcca ggtgggacca gagatatcgc tagcaaggat ccatggcatc acaggtcgtc
      421 tatggggaat gggcatcaac tcacaccgag tgctacaata tgtccagaat cgactccacc
      481 ttcgtctcac tgctgcagct ggtgtgggcc gtggtctcag gctgccagac aatcttcatg
      541 attagcaggg ctcccaaggt cccttgggag tccgtgtacc tgccctttgt cgaatctatc
      601 acttatgcac tggccagtac tggcaacgga accctgcaga tgagggacgg cagattcttt
      661 ccttggagcc gaatggcctc ctggctgtgc acctgtccaa tcatgctggg acagatttct
      721 aacatggctc tggtgaagta caaaagtatc ccactgaatc ccattgcaca ggccgcttcc
      781 atcattagag tggtcatggg aatcaccgct acaatttctc ccgcagagta tatgaagtgg
      841 ctgttctttt tctttggggc cacctgcctg gtgttcgagt acagtgtggt cttcacaatc
      901 tttcaagtgg ggctgtatgg ctttgaatct gtcgggacac ctctggcaca gaaggtggtg
      961 gtgaggatca agatgctgcg cctgatcttc tttattgctt ggactatgtt cccaatcgtg
     1021 tggctgattt cacccaccgg agtctgcgtg atccacgaga acgtgagcgc cattctgtac
     1081 ctgctggctg acggcctgtg caaaaataca tatggagtga tcctgtggag cactgcctgg
     1141 ggagtgctgg aagggaagtg ggaccccgca tgtctgccag ggcaggagaa acccgaagcc
     1201 gacgatcctt ttggcctgaa tcacgagaag aatgcccctc ctaacgacga agtcaatatc
     1261 agaatgtttg gaaggggacg tacggcggcc aagagcagga tcaccagcga gggcgagtac
     1321 atccccctgg accagatcga catcaacgtg gcggtacccg tgagcaaggg cgaggagctg
     1381 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1441 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1501 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     1561 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1621 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1681 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1741 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1801 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     1861 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1921 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     1981 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2041 gggatcactc tcggcatgga cgagctgtac aagaccggtc agagccagcc catcctgaac
     2101 accaaggaga tggcccccca gagcaagccc cccgaggagc tggagatgag cagcatgccc
     2161 agccccgtgg cccccctgcc cgcccgcacc gagggcgtga tcgacatgcg cagcatgagc
     2221 agcatcgaca gcttcatcag ctgcgccacc gacttccccg aggccacccg cttctgatct
     2281 agaaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2341 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2401 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2461 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     2521 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     2581 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     2641 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     2701 tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac
     2761 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     2821 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     2881 ccgcatcgat accgagcgct gctcgagaga tctgtgatag cggccatcaa gctggccgcg
     2941 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3001 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3061 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3121 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cagcttatcg
     3181 ataccgcatg cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     3241 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     3301 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     3361 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     3421 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3481 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     3541 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3601 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     3661 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3721 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     3781 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     3841 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     3901 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     3961 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4021 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     4081 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4141 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4201 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4261 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4321 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4381 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4441 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4501 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4561 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4621 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4681 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4741 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4801 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4861 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4921 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4981 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5041 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5101 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5161 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5221 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5281 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5341 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5401 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5461 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5521 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5581 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5641 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5701 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5761 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5821 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5881 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5941 gctcacatgt cct
//
