LOCUS       sequence-480864-        6948 bp    DNA     circular SYN 28-JUL-2025
DEFINITION  AAV transfer plasmid for CAG-mediated co-expression of jRGECO1 and 
            mGreenLantern.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6948)
  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 6948)
  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-jRGECO1-P2A-mGreenLantern
FEATURES             Location/Qualifiers
     source          1..6948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..30
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             519..575
                     /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    1332..1920
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1385..1405)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1803..1814)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1963..2084
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1974..1993)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2028..2047
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2246..2375
                     /label=AAV2 ITR
     rep_origin      complement(2612..3200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2692..2711)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3371..4162,4163..4231))
                     /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     3994..4013
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4232..4336)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4362..4817)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4499..4520)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4711..4730
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   4889..5018
                     /label=AAV2 ITR
     promoter        5329..5606
                     /label=chicken beta-actin promoter
     primer_bind     5885..5907
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     5969..5988
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             6099..6116
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6120..6152
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(6156..6164,6165..6179)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             6207..6263
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
ORIGIN
        1 cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggctacg cgtgacgacc
       61 tgactgaaga gcagatcgca gaatttaaag aggctttctc cctatttgac aaggacgggg
      121 atgggacgat aacaaccaag gagcttggga cggtgttccg atcgctgggg cagaacccca
      181 cagaagcaga gctgcaggac atgatcaatg aagtagatgc cgacggtgac ggcacattcg
      241 acttccctga gttcctgacg atgatggcaa gaaaaatgaa tgacacagac agtgaagagg
      301 aaattcgcga agcgttccgc gtgtttgata aggacggcaa tggctacatc ggcgcagcag
      361 agcttcgcca cgtgatgaca gaccttggag agaagttaac agatgaggag gttgatgaaa
      421 tgatcagggt agcagacatc gatggggatg gtcaggtaaa ctacgaagag tttgtacaaa
      481 tgatgacagc gaagcgaccg gttcacgtgg gaagtggggc tacaaacttc tcactgctga
      541 aacaggccgg ggatgtggag gaaaacccag gaccacacgt gactagtatg gtctccaagg
      601 gtgaagagct gttcactggc gtggtgccta tcttggtgga actcgacggt gatgtgaacg
      661 gccataagtt ctccgtcaga ggtgaaggcg agggtgacgc tactaacggc aagttgacac
      721 tgaagttcat ctgcactaca ggtaagctgc ctgtgccatg gccaacactg gtgacaacct
      781 tgggctatgg tgtggcctgc tttgctagat atccagatca catgaagcag catgacttct
      841 tcaagtccgc aatgcctgag ggatacgtgc aagagcggac tatctctttc aaagacgacg
      901 gcacctacaa gacaagggca gaagtgaagt ttgagggtga cactctcgtg aacagaatcg
      961 tgctgaaggg catcgacttt aaagaagacg gcaacattct tggccacaag ctggagtata
     1021 acttcaacag ccataaggtg tacatcacag ctgacaagca gaagaacggc attaaggcaa
     1081 acttcaagac caggcataac gtggaagacg gtggagtgca gctggctgat cattaccagc
     1141 agaacacacc tatcggagat ggaccagtgc ttctgcctga taaccactat ctgagccatc
     1201 agagcaagtt gtccaaagat ccaaacgaga agagagacca catggtgctg aaagagcggg
     1261 tgactgctgc tggtatcaca cacgatatgg acgagttgta caagtgagaa ttcgatatca
     1321 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     1381 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     1441 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     1501 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     1561 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     1621 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     1681 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     1741 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     1801 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     1861 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     1921 atcgataccg tcgacccggg cggccgcttc gagcagacat gataagatac attgatgagt
     1981 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     2041 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     2101 ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc
     2161 tctacaaatg tggtaaaatc gataaggatc ttcctagagc atggctacgt agataagtag
     2221 catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct
     2281 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     2341 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgca ttaatgaatc ggccaacgcg
     2401 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2461 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2521 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2581 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2641 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2701 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2761 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2821 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2881 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2941 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3001 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3061 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3121 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3181 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3241 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3301 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3361 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3421 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3481 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3541 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3601 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3661 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3721 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3781 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3841 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3901 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3961 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4021 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4081 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4141 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4201 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4261 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4321 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     4381 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4441 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4501 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4561 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4621 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4681 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4741 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4801 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     4861 ggtgcgggcc tcttcgctat tacgccagct gcgcgctcgc tcgctcactg aggccgcccg
     4921 ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg
     4981 cagagaggga gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc
     5041 atgctactta tctacgtagc catgctctag gaagagtacc attgacgtca ataatgacgt
     5101 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     5161 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     5221 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     5281 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc
     5341 ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat
     5401 ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggg cgcgcgccgg
     5461 gggggggggg gggggggggg gggsggggsg gggsgrggcg gagaggtgcg gcggcagcca
     5521 atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct
     5581 ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc
     5641 gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg
     5701 tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc
     5761 ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg
     5821 gggggagcgg ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg
     5881 gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat
     5941 gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat
     6001 tttggcaaag aattggatcc ggtaccgcca ccatgctgca gaacgagctt gctcttaagt
     6061 tggctggact tgatattaac aagactggag gaggttctca tcatcatcat catcatggta
     6121 tggctagcat gactggtgga cagcaaatgg gtcgggatct gtacgacgat gacgataagg
     6181 atctcgcaac aatggtcgac tcatcacgtc gtaagtggaa taaggcaggt cacgcagtca
     6241 gagctatagg tcggctgagc tcacccgtgg tttccgagcg gatgtacccc gaggacggcg
     6301 ccctgaagag cgagatcaag aaggggctga ggctgaagga cggcggccac tacgccgccg
     6361 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacatcgtgg
     6421 acatcaagtt ggacatcgtg tcccacaacg aggactacac catcgtggaa cagtgcgaac
     6481 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaaggga ggtacaggcg
     6541 ggagtctggt gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct
     6601 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
     6661 agggccgccc ctacgaggcc tttcagaccg ctaagctgaa ggtgaccaag ggtggccccc
     6721 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacatta
     6781 agcacccagc cgacatcccc gactacttca agctgtcctt ccccgagggc ttcaggtggg
     6841 agcgcgtgat gaacttcgag gacggcggca ttattcacgt gaaccaggac tcctccctgc
     6901 aggacggcgt attcatctac aaggtgaagc tgcgcggcac caacttcc
//
