LOCUS       sequence-478903-        6035 bp    DNA     circular SYN 17-JUL-2025
DEFINITION  An AAV genome that ubiquitously expresses the fluorescent protein 
            cyOFP from the CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6035)
  AUTHORS   Haziza S, Chrapkiewicz R, Zhang Y, Kruzhilin V, Li J, Li J, Delamare
            G, Swanson R, Buzsaki G, Kannan M, Vasan G, Lin MZ, Zeng H, Daigle 
            TL, Schnitzer MJ
  TITLE     Imaging high-frequency voltage dynamics in multiple neuron classes 
            of behaving mammals.
  JOURNAL   Cell, 188, 1-23, August 7, 2025. doi: 10.1016/j.cell.2025.06.028
  PUBMED    39185175
REFERENCE   2  (bases 1 to 6035)
  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: "Cell, 188, 
            1-23, August 7, 2025. doi: 10.1016/j.cell.2025.06.028"
COMMENT     Sequence Label: pAAV-CAG-cyOFP-WPRE
FEATURES             Location/Qualifiers
     source          1..6035
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        436..714
                     /label=chicken beta-actin promoter
     primer_bind     993..1015
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1077..1096
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1130..1831
                     /codon_start=1
                     /product="orange-red fluorescent protein that is excitable 
                     by cyan light and suitable as an acceptor for blue-emitting
                     luciferase (Chu et al., 2016)"
                     /label=CyOFP1
                     /translation="MVSKGEELIKENMRSKLYLEGSVNGHQFKCTHEGEGKPYEGKQTN
                     RIKVVEGGPLPFAFDILATHFMYGSKVFIKYPADLPDYFKQSFPEGFTWERVMVFEDGG
                     VLTATQDTSLQDGELIYNVKVRGVNFPANGPVMQKKTLGWEPSTETMYPADGGLEGRCD
                     KALKLVGGGHLHVNFKTTYKSKKPVKMPGVHYVDRRLERIKEADNETYVEQYEHAVARY
                     SNLGGGMDELYK"
     misc_feature    1855..2443
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1908..1928)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2326..2337)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2470..2487)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2476..2700
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(2910..3239)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2924..3059
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3224..3244
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3485..4073)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3565..3584)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4244..5035,5036..5104))
                     /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     4867..4886
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5105..5209)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5277..5295
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5333..5355)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5455..5474
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5688..5707
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5997..6015
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5997..6014
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggtaccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      241 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      301 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      361 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      421 tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc
      481 cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg
      541 cggggggggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg
      601 gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
      661 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
      721 gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg
      781 ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc
      841 tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt
      901 gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggctgtc cgcgggggga
      961 cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg
     1021 ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa
     1081 cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttggatccga tggtgagcaa
     1141 gggcgaggag ctgatcaagg agaacatgag aagcaagctg tacctggaag gcagcgtgaa
     1201 cggccaccag ttcaagtgca cccacgaagg ggagggcaag ccctacgagg gcaagcagac
     1261 caacaggatc aaggtggtgg agggaggccc cctgccgttc gcattcgaca tcctggccac
     1321 ccactttatg tacgggagca aggtgttcat caagtacccc gccgacctcc ccgattattt
     1381 taagcagtcc ttccctgagg gcttcacatg ggagagagtc atggtgttcg aagacggggg
     1441 cgtgctgacc gccacccagg acaccagcct ccaggacggc gagctcatct acaacgtcaa
     1501 ggtcagaggg gtgaacttcc cagccaacgg ccccgtgatg cagaagaaaa cactgggctg
     1561 ggagcccagc accgagacca tgtaccccgc tgacggcggc ctggaaggca gatgcgacaa
     1621 ggccctgaag ctcgtgggcg ggggccacct gcacgtcaac ttcaagacca catacaagtc
     1681 caagaaaccc gtgaagatgc ccggcgtcca ctacgtggac cgcagactgg aaagaatcaa
     1741 ggaggccgac aacgagacct acgtcgagca gtacgagcac gctgtggcca gatactccaa
     1801 cctgggcgga ggcatggacg agctgtacaa gtaatgaatt cgaagcttat cgataatcaa
     1861 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     1921 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     1981 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     2041 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     2101 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     2161 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2221 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     2281 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     2341 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     2401 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgactc
     2461 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2521 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     2581 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     2641 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     2701 cttctgaggc ggaaagaacc agctggggct cgactagagc atggctacgt agataagtag
     2761 catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct
     2821 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcggc
     2881 ctcagtgagc gagcgagcgc gcagagcttt ttgcaaaagc ctaggcctcc aaaaaagcct
     2941 cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa
     3001 aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg
     3061 ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt
     3121 ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg
     3181 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca
     3241 ttccacagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3301 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3361 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3421 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3481 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3541 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3601 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3661 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3721 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3781 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3841 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3901 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3961 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4021 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4081 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4141 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4201 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4261 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4321 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4381 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4441 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4501 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4561 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4621 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4681 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4741 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4801 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4861 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4921 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4981 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5041 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5101 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5161 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5221 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     5281 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     5341 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     5401 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     5461 cagagcagat tgtactgaga gtgcaccatt cgacgctctc ccttatgcga ctcctgcatt
     5521 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     5581 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     5641 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     5701 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     5761 tagaggatct ggctagcgat gaccctgctg attggttcgc tgaccatttc cgggtgcggg
     5821 acggcgttac cagaaactca gaaggttcgt ccaaccaaac cgactctgac ggcagtttac
     5881 gagagagatg atagggtctg cttcagtaag ccagatgcta cacaattagg cttgtacata
     5941 ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca catacgattt
     6001 aggtgacact atagaataca cggaattaat tctag
//
