LOCUS       sequence-479001-        6537 bp    DNA     circular SYN 18-JUL-2025
DEFINITION  AAV-vector for optogenetic activation of noradrenergic neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6537)
  AUTHORS   Wissing C, Eschholz LS, Maheu M, Sauter K, Morellini F, Wiegert S, 
            Dieter A
  TITLE     Targeting Norepinephrine Neurons of the Locus Coeruleus: A 
            Comparison of Model Systems and Strategies
  JOURNAL   bioRxiv 2022.01.22.477348
REFERENCE   2  (bases 1 to 6537)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 18, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2022.01.22.477348"
COMMENT     Sequence Label: PRS_8-ChrimsonR-tdTomato
FEATURES             Location/Qualifiers
     source          1..6537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(365..1156,1157..1225))
                     /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     988..1007
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1226..1330)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1398..1416
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1454..1476)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1576..1595
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1612..2067)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1749..1770)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1961..1980
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2142..2282
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2153..2282
                     /label=AAV2 ITR
     polyA_signal    2290..2497
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2486..2503
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    2533..3121
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2639..2650
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3048..3068
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    3131..3155
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(4602..5651)
                     /codon_start=1
                     /product="red light-drivable channelrhodopsin from 
                     Chlamydomonas noctigama (Klapoetke et al., 2014)"
                     /label=ChrimsonR
                     /note="K176R mutant with accelerated closing kinetics"
                     /translation="MAELISSATRSLFAAGGINPWPNPYHHEDMGCGGMTPTGECFSTE
                     WWCDPSYGLSDAGYGYCFVEATGGYLVVGVEKKQAWLHSRGTPGEKIGAQVCQWIAFSI
                     AIALLTFYGFSAWKATCGWEEVYVCCVEVLFVTLEIFKEFSSPATVYLSTGNHAYCLRY
                     FEWLLSCPVILIRLSNLSGLKNDYSKRTMGLIVSCVGMIVFGMAAGLATDWLKWLLYIV
                     SCIYGGYMYFQAAKCYVEANHSVPKGHCRMVVKLMAYAYFASWGSYPILWAVGPEGLLK
                     LSPYANSIGHSICDIIAKEFWTFLAHHLRIKIHEHILIHGDIRKTTKMEIGGEEVEVEE
                     FVEEEDEDTV"
     regulatory      5648..5657
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(5658..5682)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5914..5934)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     repeat_region   5941..6081
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5941..6070
                     /label=AAV2 ITR
     rep_origin      complement(join(6143..6537,1..194))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6223..6242)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
       61 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
      121 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
      181 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
      241 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
      301 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
      361 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
      421 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
      481 gccccagtgc tgcaatgata ccgcggcttc cacgctcacc ggctccagat ttatcagcaa
      541 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
      601 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
      661 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
      721 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
      781 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
      841 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
      901 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
      961 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1021 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     1081 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     1141 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     1201 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     1261 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     1321 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     1381 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     1441 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     1501 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     1561 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat aaaattgtaa
     1621 acgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     1681 aatagaccga aatcggcaaa atcccttata aatcaaaaga atagcccgag atagagttga
     1741 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     1801 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt
     1861 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     1921 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     1981 cgggcgctaa ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2041 cgcttaatgc gccgctacag ggcgcgtact atggttgctt tgacgtatgc ggtgtgaaat
     2101 accgcacaga tgcgtaagga gaaaataccg catcaggcgc ccctgcaggc agctgcgcgc
     2161 tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc
     2221 ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc
     2281 ctgcggccct ccccagcatg cctgctattc tcttcccaat cctccccctt gctgtcctgc
     2341 cccaccccac cccccagaat agaatgacac ctactcagac aatgcgatgc aatttcctca
     2401 ttttattagg aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag
     2461 gggcaaacaa cagatggctg gcaactagaa ggcacagtcg aggctgatca gcgagctcta
     2521 ggaattcccg atgcggggag gcggcccaaa gggagatccg actcgtctga gggcgaaggc
     2581 gaagacgcgg aagaggccgc agagccggca gcaggccgcg ggaaggaagg tccgctggat
     2641 tgagggccga agggacgtag cagaaggacg tcccgcgcag aatccaggtg gcaacacagg
     2701 cgagcagcca tggaaaggac gtcagcttcc ccgacaacac cacggaattg tcagtgccca
     2761 acagccgagc ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa
     2821 tagggagggg gaaagcgaaa gtcccggaaa ggagctgaca ggtggtggca atgccccaac
     2881 cagtgggggt tgcgtcagca aacacagtgc acaccacgcc acgttgcctg acaacgggcc
     2941 acaactcctc ataaagagac agcaaccagg atttatacaa ggaggagaaa atgaaagcca
     3001 tacgggaagc aatagcatga tacaaaggca ttaaagcagc gtatccacat agcgtaaaag
     3061 gagcaacata gttaagaata ccagtcaatc tttcacaaat tttgtaatcc agaggttgat
     3121 tatcggaatt cccactttgt acaagaaagc tgggtttact tatacagctc atccatgccg
     3181 tacagaaaca gatggtggcg tccttcagac ctctcatact gttccacaat ggtgtagtct
     3241 tcattgtggg atgtgatatc cagcttggtg tccacatagt aatagccggg cagctgcact
     3301 ggtttcttgg ccatgtagat ggttttgaac tccaccaggt agtggccgcc gtctttcagc
     3361 ttcagggcct ggtggatttc gcccttcagc actccgtctc taggatacag ccgttcagtg
     3421 ctggcctccc accccattgt tttcttctgc atcacgggcc catcgggtgg aaaattggtg
     3481 ccgcgcattt tgactttgta gatcagagtg ccatcctgca gagaggagtc ctgtgtcacg
     3541 gtcaccaggc ctccatcttc aaaattcatc actctctccc acttaaagcc ctctgggaaa
     3601 gacagtttct tgtaatcagg aatgtcagcg ggatgcttga cataggcctt acttccgtac
     3661 ataaactgag gagacaggat atcccaagcg aaaggcagtg gtcctccctt ggtcactttc
     3721 agcttggcag tctgtgtgcc ctcgtaaggc ctcccttcgc cctctccttc aatctcaaat
     3781 tcgtgcccat tcatgctgcc ctccatgcgc accttaaacc tcatgaactc tttgatcaca
     3841 gccatattgt tatcctcgct tgaggcggtt ccactagacc ctgagccggt acttccagtc
     3901 ccatggccca ggaacagatg gtgtctcccc tcgctccgtt catactgctc cacaatagtg
     3961 tagtcctcgt tgtgtgaagt gatatccagt tttgtgtcca cgtaatagta gcctggcagc
     4021 tgcacaggtt tcttggccat gtagattgtc ttgaactcca ccaggtaatg tcccccatct
     4081 ttcagcttca gagcctggtg gatttctcct ttcagcaccc cgtcgcgtgg atacagcctt
     4141 tcggtggagg cctcccatcc catagttttc ttctgcatca caggtccatc agggggaaaa
     4201 tttgtccctc tcattttcac cttgtagatc agggtcccat cctgcaggga gctgtcctgg
     4261 gtcacagtca ccaggcctcc gtcctcgaag ttcatcactc tttcccactt aaacccctcg
     4321 gggaagctca gtttcttata atcgggaatg tcagctggat gtttcacata ggccttggag
     4381 ccgtacataa actgtgggct caggatgtcc caagcgaatg gcagaggccc gccctttgtc
     4441 actttcagct tggctgtctg ggtgccctcg tagggccgtc cctctccttc tccctcaatt
     4501 tcgaactcgt ggccgttcat gcttccttcc attctcacct taaaccgcat gaactctttg
     4561 atcacttcct cgcccttact cactgctact accggtgccg ccactgtgtc ctcgtcctcc
     4621 tcctccacga actcttccac ttccacctcc tcgcctccga tctccatctt ggtggtcttc
     4681 cggatgtcgc cgtggatcag gatgtgctcg tggatcttga tcctcaggtg gtgggccagg
     4741 aaggtccaaa actccttggc gatgatgtcg cagatgctgt ggccgatgct gttggcgtaa
     4801 gggctcagct tcagcagtcc ttctggtccc actgcccaga ggattgggta gctgccccaa
     4861 gaggcgaagt aagcgtaggc catcagcttc acgaccatgc ggcaatggcc tttaggcacg
     4921 ctgtggttgg cttccacgta gcacttggcg gcctggaagt acatgtagcc gccgtagatg
     4981 caagacacga tatacagcag ccacttgagc caatcggtag ccagtcctgc ggccatgccg
     5041 aacacgatca ttcccacgca agacacgatc aggcccatgg tccgcttgct gtagtcgttc
     5101 ttcaggccgc tcaggttgct cagtctgatc aggatcacgg ggcaagacag cagccactcg
     5161 aagtagcgca ggcaataggc gtggttgccg gtagacaggt acactgtggc ggggctgctg
     5221 aactccttga agatctccag ggtcacgaac agcacctcga cgcaacagac gtagacctcc
     5281 tcccaaccgc aagtggcctt ccaggcgctg aagccgtaga atgtcagcag ggcgatggcg
     5341 atgctgaaag caatccactg gcagacctgg gcgccgatct tttctcctgg tgtgcctctg
     5401 ctgtgcagcc aagcctgctt cttctccact cccacgacca ggtagccgcc tgtggcctcc
     5461 acgaagcaat atccgtatcc ggcgtcgctc agtccgtaag aagggtcaca ccaccactcg
     5521 gtgctgaagc actcgcctgt aggtgtcatt cctccacagc ccatgtcctc gtggtggtag
     5581 gggttaggcc aagggttgat gcctccggcg gcaaacagag atctggtggc gctgctgatc
     5641 agctcagcca tggtggcagc ctgctttttt gtacaaactt gtctggtccc acctggccat
     5701 ttatgccctg gtcgggggta atcacatttg tctagcgggg taatcacatt tgtctagcgc
     5761 gggtaatcac atttgtctag cgggggtaat cacatttgtc tagcgggggg taatcacatt
     5821 tgtctagcgg ggtaatcaca tttgtctagc gcgggtaatc acatttgtct agcgggggta
     5881 atcacatttg tctagcggaa gctaatacgc gttcaacttt tctatacaaa gttgtctaga
     5941 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     6001 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     6061 gagcgcgcag ctgcctgcag gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6121 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6181 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6241 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6301 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6361 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6421 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6481 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtg
//
