LOCUS       sequence-382625-        5928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses miniSOG in the cytosol of mammalian cells, joined by a T2A
            cleavage site with dTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5928)
  AUTHORS   Sessler CD, Zhou Y, Wang W, Hartley ND, Fu Z, Graykowski D, Sheng M,
            Wang X, Liu J
  TITLE     Optogenetic polymerization and assembly of electrically functional 
            polymers for modulation of single-neuron excitability.
  JOURNAL   Sci Adv. 2022 Dec 9;8(49):eade1136. doi: 10.1126/sciadv.ade1136. 
            Epub 2022 Dec 7.
  PUBMED    36475786
REFERENCE   2  (bases 1 to 5928)
  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: "Sci Adv."; 
            date: "2022-12-9"; volume: "8(49):eade1136. doi"; pages: " 
            10.1126/sciadv.ade1136. Epub 2022 Dec 7"
COMMENT     Sequence Label: pAAV-CAG-miniSOG-T2A-dTomato
FEATURES             Location/Qualifiers
     source          1..5928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   56..185
                     /label=AAV2 ITR
     promoter        498..775
                     /label=chicken beta-actin promoter
     primer_bind     1054..1076
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1138..1157
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1190..1199
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1199..1222
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1238..1555
                     /codon_start=1
                     /product="mini singlet oxygen generator, a fluorescent 
                     flavoprotein suitable for correlated light and electron 
                     microscopy (Shu et al., 2011)"
                     /label=miniSOG
                     /translation="MEKSFVITDPRLPDNPIIFASDGFLELTEYSREEILGRNGRFLQG
                     PETDQATVQKIRDAIRDQREITVQLINYTKSGKKFWNLLHLQPMRDQKGELQYFIGVQL
                     DG"
     CDS             1643..1696
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    2427..3015
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2480..2500)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2898..2909)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3058..3179
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3069..3088)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3123..3142
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3341..3470
                     /label=AAV2 ITR
     rep_origin      3542..3997
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3629..3648)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3839..3860
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4023..4127
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4128..4196,4197..4988)
                     /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(4346..4365)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5159..5747
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5648..5667
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      121 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
      241 gatcgtacca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      301 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      361 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      421 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      481 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc
      541 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg
      601 ggcggggggg gggggggggc gcgcgccrgg sggggsgggg sggggsgrgg ggsggggsgg
      661 ggsgrggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt
      721 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt
      781 cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg
      841 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
      901 ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct
      961 tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggctgt ccgcgggggg
     1021 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1081 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1141 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attggatccg ccaccatgga
     1201 ttacaaggat gacgacgata agggcggagg tggttctatg gagaaaagtt tcgtgataac
     1261 tgatccacgg ctgccagaca atcccatcat cttcgcatcc gatggcttcc tggagctgac
     1321 cgagtattcc agagaggaga tcctgggccg caatggccgc tttctgcagg gaccagagac
     1381 agaccaggcc acagtgcaga agattcgcga tgccattaga gatcagcgcg agattaccgt
     1441 gcagctgata aactacacaa aaagcgggaa gaaattctgg aacctcctgc acctccagcc
     1501 catgagggac cagaagggtg agctccagta tttcatcgga gtgcagctgg atggatcagg
     1561 aggaggctca ggaggaggaa agctgaaccc tcctgatgag agtggccccg gctgcatgag
     1621 ctgctgtgtg ctctccagat ctgagggtcg gggctctctg ctcacatgtg gcgacgtcga
     1681 ggagaatccc ggaccggccc cggggtcgac aatggtgtct aagggtgagg aagtaattaa
     1741 ggagtttatg agattcaaag ttagaatgga gggtagcatg aacgggcacg agtttgaaat
     1801 tgaaggtgag ggtgaaggaa gaccatatga gggcacacaa accgccaagc tcaaggttac
     1861 caaaggtggc ccccttcctt tcgcctggga catcctcagc cctcaattca tgtacggtag
     1921 caaggcttac gtcaaacacc ctgctgatat acctgattac aaaaaactgt catttcctga
     1981 aggatttaag tgggaacgag taatgaattt tgaggatggt gggctggtca ctgtcacgca
     2041 agactcctct ctgcaggacg gcaccctgat ttacaaggtg aagatgcgcg gaacaaattt
     2101 tccccccgat ggccccgtga tgcagaagaa gaccatgggg tgggaggcct ctacagaacg
     2161 actgtacccc cgcgatggcg ttcttaaagg cgaaatccat caggcgctta aactcaaaga
     2221 tggcgggcac tacctggttg agttcaaaac tatatatatg gccaaaaagc cggttcagct
     2281 gcctggctac tactacgtgg acaccaagtt ggacatcacg agtcacaacg aggactacac
     2341 aatcgtcgag cagtacgaac gctcagaagg tcggcatcac ctgttcttgt atggcatgga
     2401 tgagctgtat aagtaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     2461 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     2521 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     2581 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     2641 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     2701 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     2761 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     2821 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     2881 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     2941 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3001 gggccgcctc cccgcatcga taccgtcgac ccgggcggcc gcttcgagca gacatgataa
     3061 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     3121 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     3181 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt
     3241 aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatcttcct agagcatggc
     3301 tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag
     3361 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     3421 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctggcgtaat
     3481 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     3541 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3601 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3661 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3721 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3781 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3841 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3901 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3961 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     4021 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4081 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4141 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4201 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4261 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4321 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4381 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4441 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4501 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4561 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4621 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4681 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4741 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4801 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4861 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4921 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4981 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5041 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5101 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5161 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5221 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5281 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5341 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5401 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5461 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5521 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5581 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5641 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5701 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5761 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5821 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     5881 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
