LOCUS       sequence-480865-        6822 bp    DNA     circular SYN 28-JUL-2025
DEFINITION  AAV transfer plasmid for CAG-mediated co-expression of KGECO1 and 
            mGreenLantern.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6822)
  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 6822)
  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-KGECO1-P2A-mGreenLantern
FEATURES             Location/Qualifiers
     source          1..6822
                     /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"
     CDS             2293..2349
                     /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    3106..3694
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3159..3179)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3577..3588)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3737..3858
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3748..3767)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3802..3821
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4020..4149
                     /label=AAV2 ITR
     rep_origin      complement(4386..4974)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4466..4485)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5145..5936,5937..6005))
                     /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     5768..5787
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6006..6110)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6136..6591)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6273..6294)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6485..6504
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6663..6792
                     /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 ggggggggsg 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 tgggcagcgt gaagctgatc cccagcctga ccaccgtgat cctcgtgaag
     1081 tccatgctgc ggaagcggag cttcggcaac cccttcaagt ataatacgga gaccctgtac
     1141 cccgctgacg gcggcctgga aggcgcatgt gacatggccc tgaagctcgt gggcgggggc
     1201 cacctgaact gcagccttga gaccacatac agatccaaga aacccgctac gaacctcaag
     1261 atgcccggtg tctacaacgt ggaccacaga ctggaacgaa tcaaagaggc cgacgatgag
     1321 acctacgtcg agctgcacga ggtggctgtg gccagatacg tgggcctggg tggtggcgga
     1381 ggtacaggcg ggagtgtgag cgagctgatt aaggagaaca tgccaatgaa gctgtacatg
     1441 gagggcaccg tgaacaacca ccacttcaag tgcacatccg agggcgaagg caagccctac
     1501 gagggcaccc agaccatgag aatcaaggtc gtcgagggcg gccctctccc cttcgccttc
     1561 gacatcctgg ctaccagctt catgtacggc agcagaacct tcatcaagca ccctcctggc
     1621 atccccgact tctttaagca gtccttccct gagggcttca catgggagag agtcaccaca
     1681 tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc
     1741 atctacaacg tcaaggttag agggatgaac ttcccagcca acggccctgt gatgcagaag
     1801 aaaacactcg gctgggaggc cagtaatggc caactgactg aagagcagat cgcagaattt
     1861 aaagaggctt tctccctatt tgacaaggac ggggatggga cgataacaac caaggagctg
     1921 gggacggtga tgcggtctct ggggcagaac cccacagaag cggagctgca ggacatgatc
     1981 aatgaagtag atgccgacgg tgacggcaca ttcgacttcc ctgagttcct gacgatgatg
     2041 gcaagaaaaa tgagttatag agtcactgaa gaggaaatta gagaagcgtt ccgcgtgttt
     2101 gataaggacg gcaatggcta catcggcgca gcagagcttc gccacgtgat gacagacctt
     2161 ggagagaagt taacagatga ggaggttgat gaaatgatca gggtagcaga catcgatggg
     2221 gatggtcagg taaactacga agagtttgta caaatgatga cagcgaagcg accggttcac
     2281 gtgggaagtg gggctacaaa cttctcactg ctgaaacagg ccggggatgt ggaggaaaac
     2341 ccaggaccac acgtgactag tatggtctcc aagggtgaag agctgttcac tggcgtggtg
     2401 cctatcttgg tggaactcga cggtgatgtg aacggccata agttctccgt cagaggtgaa
     2461 ggcgagggtg acgctactaa cggcaagttg acactgaagt tcatctgcac tacaggtaag
     2521 ctgcctgtgc catggccaac actggtgaca accttgggct atggtgtggc ctgctttgct
     2581 agatatccag atcacatgaa gcagcatgac ttcttcaagt ccgcaatgcc tgagggatac
     2641 gtgcaagagc ggactatctc tttcaaagac gacggcacct acaagacaag ggcagaagtg
     2701 aagtttgagg gtgacactct cgtgaacaga atcgtgctga agggcatcga ctttaaagaa
     2761 gacggcaaca ttcttggcca caagctggag tataacttca acagccataa ggtgtacatc
     2821 acagctgaca agcagaagaa cggcattaag gcaaacttca agaccaggca taacgtggaa
     2881 gacggtggag tgcagctggc tgatcattac cagcagaaca cacctatcgg agatggacca
     2941 gtgcttctgc ctgataacca ctatctgagc catcagagca agttgtccaa agatccaaac
     3001 gagaagagag accacatggt gctgaaagag cgggtgactg ctgctggtat cacacacgat
     3061 atggacgagt tgtacaagtg agaattcgat atcaagctta tcgataatca acctctggat
     3121 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     3181 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3241 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     3301 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     3361 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     3421 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3481 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     3541 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     3601 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     3661 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc cgggcggccg
     3721 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     3781 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3841 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3901 gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     3961 gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa
     4021 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     4081 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     4141 agcgcgcagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4201 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4261 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4321 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4381 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4441 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4501 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4561 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4621 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4681 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4741 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4801 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4861 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4921 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4981 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5041 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5101 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5161 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5221 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5281 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5341 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5401 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5461 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5521 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5581 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5641 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5701 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5761 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5821 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5881 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5941 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6001 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6061 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6121 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     6181 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     6241 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     6301 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     6361 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     6421 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     6481 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     6541 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
     6601 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     6661 agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt cgggcgacct
     6721 ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca
     6781 ctaggggttc cttgtagtta atgattaacc cgccatgcta ct
//
