LOCUS       Exported                6845 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  FLinChR is the fusion signal sequence and transmembrane (TM) domain 
            of Neurexin 1B-delta to ChR2E123T/T159C for inversion ChR2 to an 
            optogenetic inhibitor.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-CAG-FLInChR-mVenus
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6845)
  AUTHORS   Brown J, Behnam R, Coddington L, Tervo DGR, Martin K, Proskurin M, 
            Kuleshova E, Park J, Phillips J, Bergs ACF, Gottschalk A, Dudman JT,
            Karpova AY
  TITLE     Expanding the Optogenetics Toolkit by Topological Inversion of 
            Rhodopsins.
  JOURNAL   Cell. 2018 Oct 16. pii: S0092-8674(18)31238-8. doi: 
            10.1016/j.cell.2018.09.026.
  PUBMED    30343901
REFERENCE   2  (bases 1 to 6845)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6845
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        443..720
                     /label=chicken beta-actin promoter
     primer_bind     999..1021
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1083..1102
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1318..1347
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1597..2523
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYATWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSDIGCIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             2548..3264
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2590..2611)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2851..2870)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3198..3219
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3289..3877
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3342..3362)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3760..3771)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3920..4041
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3931..3950)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3985..4004
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4203..4332
                     /label=AAV2 ITR
     primer_bind     complement(4398..4415)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4569..5157)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4649..4668)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5328..6188)
                     /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     5951..5970
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6189..6293)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6319..6774)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6456..6477)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6668..6687
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatcg taccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      241 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      301 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      361 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      421 ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc
      481 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
      541 atgggggcgg gggggggggg ggggcgcgcg ccrggsgggg sggggsgggg sgrggggsgg
      601 ggsggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      661 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      721 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
      781 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      841 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      901 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gctgtccgcg
      961 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
     1021 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
     1081 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattgg atccgccacc
     1141 atgtaccaga ggatgctccg gtgcggtgcc gagctgggat cgcccggagg cggcagtagt
     1201 ggcggcgcag gggggcgcct ggccctgctc tggatagtcc cgctcaccct cagcggcctc
     1261 ctaggagtgg cctggggggc atccagtttg ggagcgcacc acatccacca taaacttgaa
     1321 caaaagctga tttctgaaga agacctcggt gggttagcca accccacccg agtgggcggc
     1381 cgcgaaccat acccaggctc ggcagaggtg atccgggagt ctagcagtac cactggcatg
     1441 gtggtgggga ttgtcgcagc agcagctctg tgcatcctca tcctcctcta tgccatgaag
     1501 aagcgcaggg ccaagggagg ttcaggaggt tcaggaggtc tcgagcacgg aactatccca
     1561 ttcaacagaa ctcacaggtc caagagaagc tccggaatgg actatggcgg cgctttgtct
     1621 gccgtcggac gcgaactttt gttcgttact aatcctgtgg tggtgaacgg gtccgtcctg
     1681 gtccctgagg atcaatgtta ctgtgccgga tggattgaat ctcgcggcac gaacggcgct
     1741 cagaccgcgt caaatgtcct gcagtggctt gcagcaggat tcagcatttt gctgctgatg
     1801 ttctatgcct accaaacctg gaaatctaca tgcggctggg aggaaatcta tgtgtgcgcc
     1861 attgaaatgg ttaaggtgat tctggagttc ttttttgagt ttaagaatcc ctctatgctc
     1921 taccttgcca caggacaccg ggtgcagtgg ctgcgctatg caacctggct gctcacttgt
     1981 cctgtcatcc ttatccacct gagcaacctc accggcctga gcaacgacta cagcaggaga
     2041 accatgggac tccttgtctc agacatcggg tgcatcgtgt ggggggctac cagcgccatg
     2101 gcaaccggct atgttaaagt catcttcttt tgtcttggat tgtgctatgg cgcgaacaca
     2161 ttttttcacg ccgccaaagc atatatcgag ggttatcata ctgtgccaaa gggtcggtgc
     2221 cgccaggtcg tgaccggcat ggcatggctg tttttcgtga gctggggtat gttcccaatt
     2281 ctcttcattt tggggcccga aggttttggc gtcctgagcg tctatggctc caccgtaggt
     2341 cacacgatta ttgatctgat gagtaaaaat tgttgggggt tgttgggaca ctacctgcgc
     2401 gtcctgatcc acgagcacat attgattcac ggagatatcc gcaaaaccac caaactgaac
     2461 atcggcggaa cggagatcga ggtcgagact ctcgtcgaag acgaagccga ggccggagcc
     2521 gtgccaggag gttcaggcgg aaccggtgtg agcaagggcg aggagctgtt caccggggtg
     2581 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     2641 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc
     2701 aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcct gcagtgcttc
     2761 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     2821 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     2881 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     2941 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3001 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     3061 gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3121 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaagctgag caaagacccc
     3181 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3241 ggcatggacg agctgtacaa gtaagaattc gatatcaagc ttatcgataa tcaacctctg
     3301 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     3361 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     3421 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     3481 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     3541 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     3601 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     3661 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     3721 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     3781 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     3841 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg acccgggcgg
     3901 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     3961 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     4021 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     4081 ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     4141 aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta
     4201 caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     4261 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     4321 gcgagcgcgc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4381 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4441 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4501 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4561 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4621 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4681 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4741 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4801 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4861 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4921 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4981 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5041 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5101 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5161 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5221 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5281 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5341 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5401 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5461 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5521 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5581 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5641 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5701 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5761 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5821 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5881 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5941 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6001 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6061 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6121 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6181 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6241 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6301 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     6361 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     6421 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     6481 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     6541 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     6601 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     6661 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     6721 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt
     6781 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     6841 gccag
//
