LOCUS       sequence-478973-        7204 bp    DNA     circular SYN 17-JUL-2025
DEFINITION  expresses Flpo recombinase-dependent GtACR1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7204)
  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 7204)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2018-08-9"; volume: "7. pii: 38506. doi"; pages: " 
            10.7554/eLife.38506"
COMMENT     Sequence Label: pAAV-EF1-alpha-FRT-FLEX-GtACR1-EYFP
FEATURES             Location/Qualifiers
     source          1..7204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1270..1303
                     /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(join(1310..2023,2024..2026))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1355..1376)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1704..1723
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1963..1984
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(3041..3074)
                     /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    3101..3689
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3154..3174)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3572..3583)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3721..4197
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3920..3939)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4237..4377
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4237..4366
                     /label=AAV2 ITR
     rep_origin      4452..4907
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4539..4558)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4749..4770
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4924..4943)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5043..5065
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5103..5121)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5189..5293
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5294..5362,5363..6154)
                     /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(5512..5531)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6325..6913
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6814..6833
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6975..7104
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccgaagt tcctattcta
     1201 tcagaagtat aggaacttcg cagaatggta gctggattgt agctgctatt agcaatatga
     1261 aacctcttag aagttcctat tctctagaaa gtataggaac ttcgaattct tacttgtaca
     1321 gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat
     1381 cgcgcttctc gttggggtct ttgctcaggg cggactggta gctcaggtag tggttgtcgg
     1441 gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca
     1501 cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct
     1561 tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga
     1621 tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc
     1681 cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc
     1741 gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt
     1801 cggggtagcg ggcgaagcac tgcaggccgt agccgaaggt ggtcacgagg gtgggccagg
     1861 gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat
     1921 cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga
     1981 ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcacggcg gccgcggtct
     2041 cgccggctct ggcgtcctcg tacctctcgg tgttgccctt ctccaggtcc tgctcgtact
     2101 caggcatcag ggtgccgtcc acgttcctgc ccttcacgta atccctgtcc cacttgccgt
     2161 tcagcaggcc ccaggttgta gcccacagca ggatgccgta ggtgttcttg cacagagcgt
     2221 cgcccagcag gtacagcacg ctgctggtgt tctcgctcat gatgcaggcg ccggtgctgg
     2281 agaagctcca caggataggg taggccatcc agctcacgta gaacacgatc ctcatcagct
     2341 tcagcctctc caccaccttc agggacatgg gggagccgat ggtctggaaa tcgtggatgg
     2401 taatggcgaa aatggcgaag gccaccacca tctcgaagat gaagaagatg ctgctgatga
     2461 aacagtacag ccacacgtgc agcgggtcta gcaccacgga ggcggtgatg ccgaacacgg
     2521 tgcagatgct ggaggcggcg atcatcatag ggttgagggg gatgctcttg tacttcacga
     2581 gggccatatt gctcaccagg cccagcatga tagggcaggt gcacagccag ctggccattc
     2641 tggcccaggg caggtacttg ccattggcca ctctgatgta gccgtttccg gtgaaggcca
     2701 gggaataggt gatcatctcg gtggtgggca ggtagatggc ttcccagggc accttggggg
     2761 ctctggtcac catgaagaac acctgacagg cgctcacgac ggccatcacc agctggacgt
     2821 acttgatgcc gtccagggtg atcaggctct tctccacgca gaactggttc tccctgctcc
     2881 attcgccgta gatggcggga tcacaggtga tgctgctcat ggatcccggc tagctacgta
     2941 ggcgcgcctt aattaagaag ttcctatact tctgatagaa taggaacttc ttgccttaac
     3001 ccagaaatta tcactgttat tctttagaat ggtgcaaaga gaagttccta tactttctag
     3061 agaataggaa cttccagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa
     3121 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3181 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3241 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3301 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3361 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3421 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3481 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat
     3541 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3601 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3661 tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg agagatctac
     3721 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     3781 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     3841 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     3901 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     3961 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     4021 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     4081 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     4141 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     4201 attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     4261 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     4321 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     4381 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     4441 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4501 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     4561 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4621 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     4681 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4741 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     4801 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     4861 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     4921 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     4981 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     5041 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     5101 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     5161 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5221 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     5281 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     5341 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     5401 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     5461 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     5521 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     5581 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5641 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5701 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5761 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5821 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     5881 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     5941 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6001 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6061 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6121 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     6181 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6241 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6301 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6361 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6421 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6481 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6541 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6601 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6661 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6721 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6781 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6841 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6901 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6961 ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc
     7021 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg
     7081 ccaactccat cactaggggt tcctgcggcc gcacgcgtaa gctttgcaaa gatggataaa
     7141 gttttaaaca gagaggaatc tttgcagcta atggaccttc taggtcttga aaggagtggg
     7201 aatt
//
