LOCUS       sequence-485513-        6725 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of FtACR-EYFP in cells under CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6725)
  AUTHORS   Govorunova EG, Sineshchekov OA, Li H, Gou Y, Chen H, Yang S, Wang Y,
            Mitchell S, Palmateer A, Brown LS, St-Pierre F, Xue MG, Spudich JL
  TITLE     Blue-shifted ancyromonad channelrhodopsins for multiplex 
            optogenetics.
  JOURNAL   Elife. 2025 Sep 19;14:RP106508. doi: 10.7554/eLife.106508.
  PUBMED    40970384
REFERENCE   2  (bases 1 to 6725)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2025-09-19"; volume: "14:RP106508. doi"; pages: " 
            10.7554/eLife.106508"
COMMENT     Sequence Label: pAAV-CAG(short)-FtACR-EYFP-WPRE-hGH
FEATURES             Location/Qualifiers
     source          1..6725
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        210..513
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        515..792
                     /label=chicken beta-actin promoter
     primer_bind     1122..1144
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1206..1225
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(2093..2095,2096..2809)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2135..2156)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2396..2415)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2743..2764
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2852..3440
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2905..2925)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3323..3334)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3472..3948
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3671..3690)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3988..4128
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3988..4117
                     /label=AAV2 ITR
     rep_origin      4203..4658
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4290..4309)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4500..4521
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4675..4694)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4794..4816
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4854..4872)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4940..5044
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5045..5113,5114..5905)
                     /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(5263..5282)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6076..6664
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6565..6584
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaacact agttattaat agtaatcaat tacggggtca
      181 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      241 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      301 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      361 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      421 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      481 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
      541 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
      601 ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccaggggg gggggggsgg
      661 ggsgaggggs ggggsggggs gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg
      721 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
      781 gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc
      841 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac
      901 ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg
      961 tggctgcgtg aaagccttga ggggctccgg gagcgccggc aggaaggaaa tgggcgggga
     1021 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1081 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1141 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1201 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgcggcccaa
     1261 cggtaccgcg ggcccgggat ccatggtgtc cggcgaggat aatgtgtcta tcgcctacgc
     1321 catgatggtc ctggtgtctg gcatcagcgt ggtccaggcc atctggtaca ccatccaaac
     1381 agccagaggc aagaaccaca gctggcggcc agctctggtc atcggcgctg aggcctttca
     1441 gtacatggtg cggctggccg tgtccctgac cgacagcaac atctccagcg gactgaccgt
     1501 gatgaacagc agaggcctgg aagtggatat cggcacctac ctgggctggt tcttcacctg
     1561 tcctctgctg gtggacgcca ttggcaatgt ggaggacgac atcatctaca gcgttgagct
     1621 gtaccgggcc gccatgacca acaccgaggc cgtgtacagc cctagaagag tgtggtggat
     1681 gagcgccgtg atgtctggca tgctgtgtac agtggtggtg gcctctctgg ctgaaaagac
     1741 cacccagatc gccgctgtct tttgcgtggg cggagccctg tgcatggtgc tcttcgccgg
     1801 catcggaaga gccattagac ggtgcctcga ccacggcgct gatagactga cagcatatag
     1861 cctgctgatc gtgttctaca caggctggag cctgttcccc ctgatctggg tgctttctcc
     1921 tctgggatac gacgtgctgg atgccgtgac cgccagggcc ctgttcggct tcgccgacct
     1981 gatcgccaaa aacctgctga gcgtgctgct gacaagactg accgtgatga gcgtgaacgc
     2041 caagcaggct gtgctggaac tgaagatcca gacaagcagc cacagcgtta acgtgagcaa
     2101 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     2161 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     2221 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     2281 cttcggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     2341 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     2401 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     2461 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     2521 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     2581 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     2641 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     2701 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     2761 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtgag ctagcgaatt
     2821 caagcttaaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga
     2881 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2941 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     3001 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     3061 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     3121 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     3181 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     3241 ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg
     3301 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3361 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3421 cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat
     3481 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     3541 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     3601 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     3661 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     3721 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc
     3781 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     3841 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     3901 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag
     3961 gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     4021 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     4081 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     4141 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     4201 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4261 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4321 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4381 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     4441 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4501 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     4561 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     4621 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     4681 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     4741 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4801 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     4861 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     4921 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4981 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5041 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5101 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5161 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5221 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5281 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5341 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5401 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5461 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5521 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5581 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5641 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5701 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5761 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5821 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5881 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5941 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6001 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6061 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6121 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6181 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6241 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6301 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6361 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6421 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6481 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6541 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6601 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6661 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6721 catgt
//
