LOCUS       Exported                7655 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Archaerhodopsin fused to mRuby2 and targeted to the neuronal soma 
            and proximal dendrites.
ACCESSION   .
VERSION     .
KEYWORDS    pCAG-eArch3.0-mRuby2-ST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7655)
  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 7655)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7655
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             1744..2517
                     /codon_start=1
                     /product="light-driven proton pump archaerhodopsin-3 from 
                     Halorubrum sodomense (Chow et al., 2010)"
                     /label=Arch
                     /translation="MDPIALQAGYDLLGDGRPETLWLGIGTLLMLIGTFYFLVRGWGVT
                     DKDAREYYAVTILVPGIASAAYLSMFFGIGLTEVTVGGEMLDIYYARYADWLFTTPLLL
                     LDLALLAKVDRVTIGTLVGVDALMIVTGLIGALSHTAIARYSWWLFSTICMIVVLYFLA
                     TSLRSAAKERGPEVASTFNTLTALVLVLWTAYPILWIIGTEGAGVVGLGIETLLFMVLD
                     VTAKVGFGFILLRSRAILGDTEAPEPSAGADVSAAD"
     CDS             2599..3309
                     /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          3551..3683
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3666..3685
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3692..4280
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3745..3765)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4163..4174)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4307..4324)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4313..4424
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4520..4540)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4520..4538)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4559..4575)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4677..4696)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4742..4797
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4796..4815)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5158..5174)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5182..5198
                     /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(5206..5236)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5251..5272
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5331..5526
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5377..5512
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5439..5458
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5532..5666
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5569..5588)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5623..5642
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5734..5751)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5905..6493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5985..6004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6664..7524)
                     /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     7287..7306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7525..7629)
                     /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 gccgggcggg 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 tcggtaccta
     1741 gcaatggacc ccatcgctct gcaggctggt tacgacctgc tgggtgacgg cagacctgaa
     1801 actctgtggc tgggcatcgg cactctgctg atgctgattg gaaccttcta ctttctggtc
     1861 cgcggatggg gagtcaccga taaggatgcc cgggaatatt acgctgtgac tatcctggtg
     1921 cccggaatcg catccgccgc atatctgtct atgttctttg gtatcgggct tactgaggtg
     1981 accgtcgggg gcgaaatgtt ggatatctat tatgccaggt acgccgactg gctgtttacc
     2041 accccacttc tgctgctgga tctggccctt ctcgctaagg tggatcgggt gaccatcggc
     2101 accctggtgg gtgtggacgc cctgatgatc gtcactggcc tcatcggagc cttgagccac
     2161 acggccatag ccagatacag ttggtggttg ttctctacaa tttgcatgat agtggtgctc
     2221 tattttctgg ctacatccct gcgatctgct gcaaaggagc ggggccccga ggtggcatct
     2281 acctttaaca ccctgacagc tctggtcttg gtgctgtgga ccgcttaccc tatcctgtgg
     2341 atcataggca ctgagggcgc tggcgtggtg ggcctgggca tcgaaactct gctgtttatg
     2401 gtgttggacg tgactgccaa ggtcggcttt ggctttatcc tgttgagatc ccgggctatt
     2461 ctgggcgaca ccgaggcacc agaacccagt gccggtgccg atgtcagtgc cgccgacctc
     2521 gagaagagca ggatcaccag cgagggcgag tacatccccc tggaccagat cgacatcaac
     2581 gtggcggccg caccggtaat ggtgtctaag ggcgaagagc tgatcaagga aaatatgcgt
     2641 atgaaggtgg tcatggaagg ttcggtcaac ggccaccaat tcaaatgcac aggtgaagga
     2701 gaaggcaatc cgtacatggg aactcaaacc atgaggatca aagtcatcga gggaggaccc
     2761 ctgccatttg cctttgacat tcttgccacg tcgttcatgt atggcagccg tacttttatc
     2821 aagtacccga aaggcattcc tgatttcttt aaacagtcct ttcctgaggg ttttacttgg
     2881 gaaagagtta cgagatacga agatggtgga gtcgtcaccg tcatgcagga caccagcctt
     2941 gaggatggct gtctcgttta ccacgtccaa gtcagagggg taaactttcc ctccaatggt
     3001 cccgtgatgc agaagaagac caagggttgg gagcctaata cagagatgat gtatccagca
     3061 gatggtggtc tgaggggata cactcatatg gcactgaaag ttgatggtgg tggccatctg
     3121 tcttgctctt tcgtaacaac ttacaggtca aaaaagaccg tcgggaacat caagatgccc
     3181 ggtatccatg ccgttgatca ccgcctggaa aggttagagg aaagtgacaa tgaaatgttc
     3241 gtagtacaac gcgaacacgc agttgccaag ttcgccgggc ttggtggtgg gatggacgag
     3301 ctgtacaagg ccctagagca gtcccagccc atcctcaaca ccaaggagat ggccccgcag
     3361 agcaagcctc cagaggagct ggagatgagc agcatgccca gccccgtggc ccctctgccc
     3421 gcacgcacgg agggcgtcat cgacatgcgg agcatgtcca gcattgacag cttcatcagc
     3481 tgtgccacgg acttccctga agccaccaga ttcttctgct acgagaacga ggtgtaactc
     3541 gaggccgcag gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg
     3601 cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat
     3661 ccactttgcc tttctctcca caggtgtcga caatcaacct ctggattaca aaatttgtga
     3721 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     3781 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     3841 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     3901 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     3961 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     4021 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     4081 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     4141 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     4201 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     4261 cctttgggcc gcctccccgc ctggaattcg agctcggtac gatcagcctc gactgtgcct
     4321 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     4381 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     4441 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     4501 aatagcccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     4561 catagctgtt tcctgtgtga aattgttatc cgctaattca ctcctcaggt gcaggctgcc
     4621 tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt
     4681 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     4741 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     4801 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     4861 ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat
     4921 cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga
     4981 ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt
     5041 tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct
     5101 gtccctcttc tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc
     5161 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5221 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5281 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat
     5341 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     5401 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     5461 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     5521 tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5581 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5641 tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag
     5701 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5761 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5821 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5881 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5941 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6001 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6061 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6121 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6181 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6241 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6301 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6361 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6421 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6481 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6541 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6601 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6661 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6721 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6781 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6841 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6901 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6961 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7021 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7081 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7141 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7201 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7261 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7321 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7381 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7441 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7501 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7561 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7621 ggttccgcgc acatttcccc gaaaagtgcc acctg
//
