LOCUS       Exported                7487 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  expresses Flpo recombinase-dependent GtACR2-Kv2.1C-linker-TlcnC, 
            which targets GtACR2 to the somatodendritic compartment. Messier et 
            al found this hybrid construct was the most effective at targeting.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-EF1-alpha-FRT-FLEX-GtACR2-EYFP-Kv2.1C-linker-TlcnC
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7487)
  AUTHORS   Messier JE, Chen H, Cai ZL, Xue M
  TITLE     Targeting light-gated chloride channels to neuronal somatodendritic 
            domain reduces their excitatory effect in the axon.
  JOURNAL   Elife. 2018 Aug 9;7. pii: 38506. doi: 10.7554/eLife.38506.
  PUBMED    30091701
REFERENCE   2  (bases 1 to 7487)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7487
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   115..244
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        345..1523
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          576..1514
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1471..1491
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1614..1647
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(1952..2665)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1994..2015)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2343..2362
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2602..2623
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(3668..3701)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    3728..4316
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3781..3801)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4199..4210)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4348..4824
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4547..4566)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4864..5004
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4864..4993
                     /label=AAV2 ITR
     rep_origin      5079..5534
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5166..5185)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5376..5397
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5551..5570)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5670..5692
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5730..5748)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5816..5920
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5921..6781
                     /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(6139..6158)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6952..53
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7441..7460
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
       61 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtcctgca
      121 ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc
      181 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt
      241 tcctgcggcc gcacgcgtaa gctttgcaaa gatggataaa gttttaaaca gagaggaatc
      301 tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
      361 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
      421 gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
      481 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
      541 ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg
      601 ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc acctggctgc
      661 agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt
      721 gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc
      781 gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag
      841 ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa
      901 atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg
      961 cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa
     1021 tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt
     1081 gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa
     1141 gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag
     1201 agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt
     1261 catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc tcgagctttt
     1321 ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg
     1381 agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg
     1441 ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt
     1501 ttcttccatt tcaggtgtcg tgaggtaccg aagttcctat tctatcagaa gtataggaac
     1561 ttcgcagaat ggtagctgga ttgtagctgc tattagcaat atgaaacctc ttagaagttc
     1621 ctattctcta gaaagtatag gaacttcgaa ttcttacttg tacaggcgcg cctcaggaag
     1681 atgtcagctg gatagcgaaa acctcgccat ctgcaggtga ctctgcagag cctgatcctg
     1741 aaccgctacc gcttccaaac cttgtagctt cggggaagtc ggtggcgcag gaaatgaagc
     1801 tatcgatact agacatgctt ctcatgtcaa tcacgccctc ggtcctagct ggcagaggag
     1861 ccactggaga tggcatggag ctcatttcca gttcctcagg gggtttactc tgaggggcca
     1921 tctccttagt gttcaggatg ggctgagact gcttgtacag ctcgtccatg ccgagagtga
     1981 tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt
     2041 tgctcagggc ggactggtag ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc
     2101 cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt
     2161 ggcggatctt gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt
     2221 tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt
     2281 cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc
     2341 gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg
     2401 gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact
     2461 gcaggccgta gccgaaggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg
     2521 tgcagatgaa cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca
     2581 cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg
     2641 tgaacagctc ctcgcccttg ctcacggcgg ccgccctgcc gaacattctg atgttgacct
     2701 cgtcgttggg gggggcattc ttctcgtggt tcagtccgaa ggggtcgtcg gcctcaggct
     2761 tctcctggcc agggaggcaa gccgggtccc actttccctc gagcactccc caggcggtgg
     2821 accacaggat cacgccgtag gtgttcttgc acaggccgtc ggcgagcaga tacaggatgg
     2881 cgctcacgtt ctcgtggatc acgcacacgc cggtggggga aatcagccac acgatgggga
     2941 acatggtcca ggcgatgaag aagatcagcc tcagcatctt gatcctcacc accaccttct
     3001 gggccagagg ggtgcccacg gactcgaagc cgtacagtcc cacctggaag atggtgaaca
     3061 ccacgctgta ttcgaagacc aggcaggtag cgccgaagaa gaagaacagc cacttcatgt
     3121 actcggcggg ggagatggta gcggtaatgc ccatcaccac cctgataatg ctggcggcct
     3181 gggcgatggg gttcagaggg atgctcttgt acttcaccag agccatgttg gagatctggc
     3241 ccagcataat ggggcatgtg cacagccagc tggccatcct ggaccaggga aagaacctgc
     3301 cgtccctcat ctgcagtgtg ccattgccgg tgctggccag ggcgtatgtg atggactcca
     3361 cgaagggcag gtacacgctc tcccagggca ccttgggggc cctgctgatc atgaagatgg
     3421 tctggcatcc gctcaccacg gcccacacca gctgcagcag gctcacgaag gtgctgtcga
     3481 tcctgctcat gttgtagcac tctgtgtggg tgctggccca ctcgccgtac acgacctggg
     3541 aggccatgga tcccggctag ctacgtaggc gcgccttaat taagaagttc ctatacttct
     3601 gatagaatag gaacttcttg ccttaaccca gaaattatca ctgttattct ttagaatggt
     3661 gcaaagagaa gttcctatac tttctagaga ataggaactt ccagaattcg atatcaagct
     3721 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3781 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3841 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3901 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3961 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4021 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4081 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     4141 gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4201 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4261 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     4321 ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct
     4381 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
     4441 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
     4501 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
     4561 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
     4621 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
     4681 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
     4741 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
     4801 accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc
     4861 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     4921 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     4981 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     5041 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     5101 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     5161 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     5221 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     5281 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     5341 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     5401 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     5461 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     5521 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5581 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5641 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5701 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5761 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5821 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5881 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5941 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     6001 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     6061 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     6121 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     6181 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     6241 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6301 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     6361 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6421 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6481 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6541 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6601 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6661 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6721 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6781 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6841 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6901 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6961 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     7021 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     7081 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     7141 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     7201 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     7261 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7321 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     7381 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7441 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctct
//
