LOCUS       sequence-480866-        6852 bp    DNA     circular SYN 28-JUL-2025
DEFINITION  AAV transfer plasmid for CAG-mediated co-expression of FRCaMPi and 
            mGreenLantern.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6852)
  AUTHORS   Zhou S, Zhu Q, Eom M, Fang S, Subach OM, Ran C, Alvarado JS, 
            Sunkavalli PS, Dong Y, Wang Y, Hu J, Zhang H, Wang Z, Sun X, Yang T,
            Mu Y, Yoon YG, Guo ZV, Subach FV, Piatkevich KD
  TITLE     A sensitive soma-localized red fluorescent calcium indicator for in 
            vivo imaging of neuronal populations at single-cell resolution.
  JOURNAL   PLoS Biol. 2025 Apr 29;23(4):e3003048. doi: 
            10.1371/journal.pbio.3003048. eCollection 2025 Apr.
  PUBMED    40299972
REFERENCE   2  (bases 1 to 6852)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 28, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS 
            Biol."; date: "2025-04-29"; volume: "23(4):e3003048. doi"; pages: " 
            10.1371/journal.pbio.3003048. eCollection 2025 Apr"
COMMENT     Sequence Label: pAAV-CAG-NES-FRCaMPi-P2A-mGreenLantern
FEATURES             Location/Qualifiers
     source          1..6852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     837..859
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     921..940
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             994..1020
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     1443..1462
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2323..2379
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     misc_feature    3136..3724
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3189..3209)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3607..3618)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3767..3888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3778..3797)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3832..3851
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4050..4179
                     /label=AAV2 ITR
     rep_origin      complement(4416..5004)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4496..4515)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5175..5966,5967..6035))
                     /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     5798..5817
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6036..6140)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6166..6621)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6303..6324)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6515..6534
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6693..6822
                     /label=AAV2 ITR
ORIGIN
        1 tatctacgta gccatgctct aggaagagta ccattgacgt caataatgac gtatgttccc
       61 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      121 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      181 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      241 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc
      301 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
      361 taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc gggggggggg
      421 gggggggggg gggggggggg sggggsgrgg cggrgrggkg cggcggcagc caatcagagc
      481 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
      541 cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc
      601 gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg
      661 cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct
      721 tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc
      781 ggctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt
      841 cggcttctgg cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt
      901 ctttttccta cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa
      961 agaattggat ccggtaccgc caccatgctt caacttcctc ctcttgaacg tcttactctt
     1021 tcgagatcta tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
     1081 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     1141 ggcgagggcc gcccctacga ggccttccag accgctaggc tgaaggtgac caagggtggc
     1201 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     1261 attaagcacc cagccgacat ccccgactac ttcaagctgt ccttccccga gggcttcagg
     1321 tgggagcgcg tgatgagctt cgaggacggc ggcattattc acgttaatca ggactcctcc
     1381 ctgcaggacg gcgtattcat ctacaaggtg aagctgctcg gcaccaactt cccccccgac
     1441 ggccccgtaa tgcagaagaa gaccatgggc tgggaggctt cctacgagat gactacccgt
     1501 aaccttacag atgagcagat tgcggagttc cgtgaggcct tttcgctgct tgatcgtgat
     1561 caggatggaa atatcacgtc caatgaattg ggtgtggtta tgaggtcgtt aggtctatcg
     1621 cctactgccg ccgaattaca agatatgatt aatgaggtcg atgccgatgg taatggcaca
     1681 attgatttta ccgaattttt gacaatgatg gcccggaaaa tgaaggatac cgacgacgaa
     1741 gaggaagttc gcgaagcctt taaagtcttc gataaagatg gaagtggata cattacagtc
     1801 gaggagctga ctcatgttct tacaagtctc ggtgaacgtt tgtctcgaga agaagtagcc
     1861 gatgtgatac gtgaagccga ctccgatggc gatggtgtaa tcaactacga agaattttct
     1921 cgtgtcattt cttccaaggg cggttcagga ggcagcgggt caggttcaat gaggaaaccg
     1981 ttccgtggcg cgggcaacgc tgtgcgtgct ttcagcactt ggaaaaagct agtgccgcgg
     2041 gtttccgagt ggatgtaccc cgaggacggc gccctgaaga gcgagatcag gaaggggctg
     2101 aggctgaagg acggcggcca ctacgccgcc gaggtcaaga ccacctacaa ggccaagaag
     2161 cccgtgcagc tgcccggcgc ctacatcgtc gacatcgagt tggacatctt gtcccacaac
     2221 gaggactaca ccatcgtgga acagtgcgaa cgcgccgtgg gccgccaccc caccggtggc
     2281 acggacgcgc tgtacaagcg accggttcac gtgggaagtg gggctacaaa cttctcactg
     2341 ctgaaacagg ccggggatgt ggaggaaaac ccaggaccac acgtgactag tatggtctcc
     2401 aagggtgaag agctgttcac tggcgtggtg cctatcttgg tggaactcga cggtgatgtg
     2461 aacggccata agttctccgt cagaggtgaa ggcgagggtg acgctactaa cggcaagttg
     2521 acactgaagt tcatctgcac tacaggtaag ctgcctgtgc catggccaac actggtgaca
     2581 accttgggct atggtgtggc ctgctttgct agatatccag atcacatgaa gcagcatgac
     2641 ttcttcaagt ccgcaatgcc tgagggatac gtgcaagagc ggactatctc tttcaaagac
     2701 gacggcacct acaagacaag ggcagaagtg aagtttgagg gtgacactct cgtgaacaga
     2761 atcgtgctga agggcatcga ctttaaagaa gacggcaaca ttcttggcca caagctggag
     2821 tataacttca acagccataa ggtgtacatc acagctgaca agcagaagaa cggcattaag
     2881 gcaaacttca agaccaggca taacgtggaa gacggtggag tgcagctggc tgatcattac
     2941 cagcagaaca cacctatcgg agatggacca gtgcttctgc ctgataacca ctatctgagc
     3001 catcagagca agttgtccaa agatccaaac gagaagagag accacatggt gctgaaagag
     3061 cgggtgactg ctgctggtat cacacacgat atggacgagt tgtacaagtg agaattcgat
     3121 atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     3181 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     3241 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     3301 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     3361 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     3421 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     3481 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     3541 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     3601 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3661 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     3721 ccgcatcgat accgtcgacc cgggcggccg cttcgagcag acatgataag atacattgat
     3781 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     3841 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     3901 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa
     3961 aacctctaca aatgtggtaa aatcgataag gatcttccta gagcatggct acgtagataa
     4021 gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc
     4081 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     4141 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcattaatg aatcggccaa
     4201 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4261 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4321 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4381 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4441 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4501 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4561 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4621 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4681 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4741 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4801 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4861 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4921 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4981 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5041 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5101 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5161 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5221 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5281 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5341 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5401 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5461 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5521 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5581 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5641 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5701 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5761 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5821 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5881 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5941 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6001 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6061 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6121 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     6181 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     6241 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     6301 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     6361 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     6421 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     6481 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     6541 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     6601 atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc
     6661 gatcggtgcg ggcctcttcg ctattacgcc agctgcgcgc tcgctcgctc actgaggccg
     6721 cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag
     6781 cgcgcagaga gggagtggcc aactccatca ctaggggttc cttgtagtta atgattaacc
     6841 cgccatgcta ct
//
