LOCUS       Exported                7259 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  expresses Flpo recombinase-dependent GtACR2-Kv4.2LL, which targets 
            GtACR2 to the somatodendritic compartment.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-EF1-alpha-FRT-FLEX-GtACR2-EYFP-Kv4.2LL
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7259)
  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 7259)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7259
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        123..1301
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          354..1292
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1249..1269
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1392..1425
                     /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(1502..2215)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1544..1565)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1893..1912
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2152..2173
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(3218..3251)
                     /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    3278..3866
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3331..3351)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3749..3760)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3898..4374
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4097..4116)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4414..4554
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4414..4543
                     /label=AAV2 ITR
     rep_origin      4629..5084
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4716..4735)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4926..4947
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5101..5120)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5220..5242
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5280..5298)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5366..5470
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5471..6331
                     /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(5689..5708)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6502..7090
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6991..7010
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   7152..22
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 aactccatca ctaggggttc ctgcggccgc acgcgtaagc tttgcaaaga tggataaagt
       61 tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa
      121 ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
      181 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
      241 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
      301 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
      361 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
      421 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
      481 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
      541 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
      601 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
      661 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
      721 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
      781 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
      841 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
      901 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
      961 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     1021 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     1081 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     1141 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     1201 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     1261 cagtggttca aagttttttt cttccatttc aggtgtcgtg aggtaccgaa gttcctattc
     1321 tatcagaagt ataggaactt cgcagaatgg tagctggatt gtagctgcta ttagcaatat
     1381 gaaacctctt agaagttcct attctctaga aagtatagga acttcgaatt cttacttgta
     1441 cagtttaaac tcacgtggtt ttctccaggc agtgaagcag gtggtggtgt tgtgtctcga
     1501 acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     1561 ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc ggactggtag ctcaggtagt
     1621 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
     1681 cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt
     1741 tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt
     1801 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     1861 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
     1921 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     1981 tcatgtggtc ggggtagcgg gcgaagcact gcaggccgta gccgaaggtg gtcacgaggg
     2041 tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt
     2101 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     2161 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcacggcgg
     2221 ccgccctgcc gaacattctg atgttgacct cgtcgttggg gggggcattc ttctcgtggt
     2281 tcagtccgaa ggggtcgtcg gcctcaggct tctcctggcc agggaggcaa gccgggtccc
     2341 actttccctc gagcactccc caggcggtgg accacaggat cacgccgtag gtgttcttgc
     2401 acaggccgtc ggcgagcaga tacaggatgg cgctcacgtt ctcgtggatc acgcacacgc
     2461 cggtggggga aatcagccac acgatgggga acatggtcca ggcgatgaag aagatcagcc
     2521 tcagcatctt gatcctcacc accaccttct gggccagagg ggtgcccacg gactcgaagc
     2581 cgtacagtcc cacctggaag atggtgaaca ccacgctgta ttcgaagacc aggcaggtag
     2641 cgccgaagaa gaagaacagc cacttcatgt actcggcggg ggagatggta gcggtaatgc
     2701 ccatcaccac cctgataatg ctggcggcct gggcgatggg gttcagaggg atgctcttgt
     2761 acttcaccag agccatgttg gagatctggc ccagcataat ggggcatgtg cacagccagc
     2821 tggccatcct ggaccaggga aagaacctgc cgtccctcat ctgcagtgtg ccattgccgg
     2881 tgctggccag ggcgtatgtg atggactcca cgaagggcag gtacacgctc tcccagggca
     2941 ccttgggggc cctgctgatc atgaagatgg tctggcatcc gctcaccacg gcccacacca
     3001 gctgcagcag gctcacgaag gtgctgtcga tcctgctcat gttgtagcac tctgtgtggg
     3061 tgctggccca ctcgccgtac acgacctggg aggccatgga tcccggctag ctacgtaggc
     3121 gcgccttaat taagaagttc ctatacttct gatagaatag gaacttcttg ccttaaccca
     3181 gaaattatca ctgttattct ttagaatggt gcaaagagaa gttcctatac tttctagaga
     3241 ataggaactt ccagaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     3301 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     3361 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     3421 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     3481 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3541 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3601 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3661 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct
     3721 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3781 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3841 gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatctacggg
     3901 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     3961 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     4021 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     4081 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     4141 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
     4201 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     4261 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     4321 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
     4381 ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     4441 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     4501 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     4561 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     4621 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4681 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     4741 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4801 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     4861 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4921 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     4981 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     5041 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     5101 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     5161 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     5221 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     5281 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     5341 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5401 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5461 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5521 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5581 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5641 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5701 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5761 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5821 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5881 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5941 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6001 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6061 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6121 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6181 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6241 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6301 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6361 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6421 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6481 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6541 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6601 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6661 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6721 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6781 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6841 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6901 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6961 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7021 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7081 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7141 tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc
     7201 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcc
//
