LOCUS       Exported                7202 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  expresses Flpo recombinase-dependent GtACR2.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-EF1-alpha-FRT-FLEX-GtACR2-EYFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7202)
  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 7202)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7202
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1510..1543
                     /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(1550..2266)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1595..1616)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1944..1963
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2203..2224
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(3269..3302)
                     /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    3329..3917
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3382..3402)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3800..3811)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3949..4425
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4148..4167)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4465..4605
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4465..4594
                     /label=AAV2 ITR
     rep_origin      4680..5135
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4767..4786)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4977..4998
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5152..5171)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5271..5293
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5331..5349)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5417..5521
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5522..6382
                     /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(5740..5759)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6553..7141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7042..7061
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccgaagt tcctattcta
     1441 tcagaagtat aggaacttcg cagaatggta gctggattgt agctgctatt agcaatatga
     1501 aacctcttag aagttcctat tctctagaaa gtataggaac ttcgaattct tacttgtaca
     1561 gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat
     1621 cgcgcttctc gttggggtct ttgctcaggg cggactggta gctcaggtag tggttgtcgg
     1681 gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca
     1741 cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct
     1801 tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga
     1861 tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc
     1921 cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc
     1981 gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt
     2041 cggggtagcg ggcgaagcac tgcaggccgt agccgaaggt ggtcacgagg gtgggccagg
     2101 gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat
     2161 cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga
     2221 ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcacggcg gccgccctgc
     2281 cgaacattct gatgttgacc tcgtcgttgg ggggggcatt cttctcgtgg ttcagtccga
     2341 aggggtcgtc ggcctcaggc ttctcctggc cagggaggca agccgggtcc cactttccct
     2401 cgagcactcc ccaggcggtg gaccacagga tcacgccgta ggtgttcttg cacaggccgt
     2461 cggcgagcag atacaggatg gcgctcacgt tctcgtggat cacgcacacg ccggtggggg
     2521 aaatcagcca cacgatgggg aacatggtcc aggcgatgaa gaagatcagc ctcagcatct
     2581 tgatcctcac caccaccttc tgggccagag gggtgcccac ggactcgaag ccgtacagtc
     2641 ccacctggaa gatggtgaac accacgctgt attcgaagac caggcaggta gcgccgaaga
     2701 agaagaacag ccacttcatg tactcggcgg gggagatggt agcggtaatg cccatcacca
     2761 ccctgataat gctggcggcc tgggcgatgg ggttcagagg gatgctcttg tacttcacca
     2821 gagccatgtt ggagatctgg cccagcataa tggggcatgt gcacagccag ctggccatcc
     2881 tggaccaggg aaagaacctg ccgtccctca tctgcagtgt gccattgccg gtgctggcca
     2941 gggcgtatgt gatggactcc acgaagggca ggtacacgct ctcccagggc accttggggg
     3001 ccctgctgat catgaagatg gtctggcatc cgctcaccac ggcccacacc agctgcagca
     3061 ggctcacgaa ggtgctgtcg atcctgctca tgttgtagca ctctgtgtgg gtgctggccc
     3121 actcgccgta cacgacctgg gaggccatgg atcccggcta gctacgtagg cgcgccttaa
     3181 ttaagaagtt cctatacttc tgatagaata ggaacttctt gccttaaccc agaaattatc
     3241 actgttattc tttagaatgg tgcaaagaga agttcctata ctttctagag aataggaact
     3301 tccagaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     3361 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3421 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3481 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3541 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     3601 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     3661 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3721 gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac
     3781 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     3841 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     3901 ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc
     3961 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
     4021 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
     4081 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
     4141 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
     4201 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag
     4261 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata
     4321 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa
     4381 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta
     4441 accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     4501 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     4561 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     4621 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     4681 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4741 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4801 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4861 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     4921 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4981 actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata
     5041 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5101 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     5161 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     5221 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     5281 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     5341 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     5401 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5461 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5521 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5581 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5641 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5701 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5761 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     5821 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5881 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5941 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6001 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6061 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     6121 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6181 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6241 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6301 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6361 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6421 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6481 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6541 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6601 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6661 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     6721 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6781 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6841 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6901 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6961 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7021 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7081 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     7141 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     7201 gt
//
