LOCUS       sequence-213588-        7695 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Anion channelrhodopsin PsuACR fused to ER export signal and 
            fluorophore mRuby2 and targeted to the neuronal soma and proximal 
            dendrites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7695)
  AUTHORS   Mardinly AR, Oldenburg IA, Pegard NC, Sridharan S, Lyall EH, Chesnov
            K, Brohawn SG, Waller L, Adesnik H
  TITLE     Precise multimodal optical control of neural ensemble activity.
  JOURNAL   Nat Neurosci. 2018 Apr 30. pii: 10.1038/s41593-018-0139-8. doi: 
            10.1038/s41593-018-0139-8.
  PUBMED    29713079
REFERENCE   2  (bases 1 to 7695)
  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"; journalName: "Nat 
            Neurosci. 2018 Apr 30. pii: 10.1038/s41593-018-0139-8. doi: 
            10.1038/s41593-018-0139-8."
COMMENT     Sequence Label: pCAG-ePsuACR-mRuby2-ST
FEATURES             Location/Qualifiers
     source          1..7695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2639..3349
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     intron          3591..3723
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3706..3725
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3732..4320
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3785..3805)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4203..4214)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4347..4364)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4353..4464
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4560..4580)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4560..4578)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4599..4615)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4717..4736)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4782..4837
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4836..4855)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5198..5214)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5222..5238
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5246..5252,5253..5270,5271..5276))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5291..5312
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5371..5566
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5417..5552
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5572..5706
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5609..5628)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5663..5682
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5774..5791)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5945..6533)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6025..6044)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6704..7495,7496..7564))
                     /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     7327..7346
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7565..7669)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcgggggggg gcgggggggg
      541 ggggggggcg gggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcggtaccga
     1741 gctcggatcc atgaccacca tcagcgaagt ctgcggcgtg tgggcactgg acaaccccga
     1801 gtgtatcgaa gtgtccggca ccaacgacaa cgtgaagatg gcccagctct gcttctgcat
     1861 ggtgtgcgtc tgccagatcc tgttcatggc cagccagtac cccaaagtcg gttgggaggc
     1921 catctacctg ccctcttgcg agtgcttcct gtacggactg gcctctagcg gaaacggctt
     1981 catccagctg tacgacggca ggctgatccc ttgggctaga tacgccgctt ggatctgcac
     2041 ctgtccctct atcctgctgc agatcaacac catccacaag tgcaagatca gccacttcaa
     2101 cctgaacacc ttcatcgtgc aggccgacct gatcatgaac atcatgggcg tgacaggcgc
     2161 cctgacaaca aacatcgcct tcaagtggat ctacttcgcc atcggctgca tcctgttcat
     2221 cttcatcgtg ctggtggtgt acgacatcat gaccagcgcc gccaaggagt ggaaggctaa
     2281 gggtgactcc aagggcaacc tggtgtccac acggctgatc ctcctccgct ggatcttcat
     2341 cgtcagttgg tgcgtgtacc cactcctctg gattctgagc cctcaggcta cttgcgcagt
     2401 gtccgaagac gtgatcagcg tggcccactt catttgcgac gccttcgcca agaacatgtt
     2461 cggcttcatc atgtggcgca ccctctggag agatctggac ggccattggg acatcagcag
     2521 acactacccc cagagcagct acgctaagga cggcaaggag gaggagcaga tgacagccat
     2581 gagccagacc gacgataccg agaagcctca cagcagccag ggagcggccg caccggtaat
     2641 ggtgtctaag ggcgaagagc tgatcaagga aaatatgcgt atgaaggtgg tcatggaagg
     2701 ttcggtcaac ggccaccaat tcaaatgcac aggtgaagga gaaggcaatc cgtacatggg
     2761 aactcaaacc atgaggatca aagtcatcga gggaggaccc ctgccatttg cctttgacat
     2821 tcttgccacg tcgttcatgt atggcagccg tacttttatc aagtacccga aaggcattcc
     2881 tgatttcttt aaacagtcct ttcctgaggg ttttacttgg gaaagagtta cgagatacga
     2941 agatggtgga gtcgtcaccg tcatgcagga caccagcctt gaggatggct gtctcgttta
     3001 ccacgtccaa gtcagagggg taaactttcc ctccaatggt cccgtgatgc agaagaagac
     3061 caagggttgg gagcctaata cagagatgat gtatccagca gatggtggtc tgaggggata
     3121 cactcatatg gcactgaaag ttgatggtgg tggccatctg tcttgctctt tcgtaacaac
     3181 ttacaggtca aaaaagaccg tcgggaacat caagatgccc ggtatccatg ccgttgatca
     3241 ccgcctggaa aggttagagg aaagtgacaa tgaaatgttc gtagtacaac gcgaacacgc
     3301 agttgccaag ttcgccgggc ttggtggtgg gatggacgag ctgtacaagg ccctagagca
     3361 gtcccagccc atcctcaaca ccaaggagat ggccccgcag agcaagcctc cagaggagct
     3421 ggagatgagc agcatgccca gccccgtggc ccctctgccc gcacgcacgg agggcgtcat
     3481 cgacatgcgg agcatgtcca gcattgacag cttcatcagc tgtgccacgg acttccctga
     3541 agccaccaga ttcttctgct acgagaacga ggtgtaactc gaggccgcag gtaagtatca
     3601 aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac
     3661 tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca
     3721 caggtgtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     3781 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     3841 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     3901 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     3961 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     4021 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     4081 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     4141 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     4201 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     4261 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     4321 ctggaattcg agctcggtac gatcagcctc gactgtgcct tctagttgcc agccatctgt
     4381 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     4441 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     4501 tggggtgggg caggacagca agggggagga ttgggaagac aatagcccag cttttgttcc
     4561 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     4621 aattgttatc cgctaattca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg
     4681 tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta
     4741 tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt
     4801 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     4861 aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat
     4921 gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc
     4981 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
     5041 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
     5101 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag
     5161 atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5221 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5281 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5341 agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc
     5401 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5461 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     5521 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt
     5581 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5641 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5701 tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5761 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5821 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5881 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5941 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6001 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6061 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6121 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6181 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6241 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6301 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6361 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6421 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6481 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6541 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6601 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6661 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6721 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6781 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6841 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6901 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6961 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7021 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7081 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7141 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7201 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7261 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7321 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7381 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7441 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7501 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7561 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7621 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7681 gaaaagtgcc acctg
//
