LOCUS       sequence-323964-        5365 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry2 under the control of TetO promoter, basic vector 
            for AAV, pAAV-CMV backbone, multiclonig site at C terminal, pAAV-TRE
            backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5365)
  AUTHORS   Tanida I, Furuta Y, Yamaguchi J, Kakuta S, Oliva Trejo JA, Uchiyama 
            Y
  TITLE     Two-color in-resin CLEM of Epon-embedded cells using osmium 
            resistant green and red fluorescent proteins.
  JOURNAL   Sci Rep. 2020 Dec 14;10(1):21871. doi: 10.1038/s41598-020-78879-x.
  PUBMED    33318540
REFERENCE   2  (bases 1 to 5365)
  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 Rep."; 
            date: "2020-12-14"; volume: "10(1):21871. doi"; pages: " 
            10.1038/s41598-020-78879-x"
COMMENT     Sequence Label: pAT-mCherry2-G
FEATURES             Location/Qualifiers
     source          1..5365
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   15..144
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    179..197
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    216..234
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    253..271
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    290..308
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    327..345
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    364..382
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    401..419
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    438..456
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     478..502
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      608..617
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             614..1321
                     /codon_start=1
                     /product="derivative of the monomeric red fluorescent 
                     protein mCherry"
                     /label=mCherry2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFNWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQCRTMGWEASTERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVDIKLDILSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(759..777)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    1379..1435
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    1506..2094
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1559..1579)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1977..1988)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2126..2602
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2325..2344)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2642..2782
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2642..2771
                     /label=AAV2 ITR
     rep_origin      2857..3312
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2944..2963)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3154..3175
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3329..3348)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3448..3470
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3508..3526)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3594..3698
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3699..3767,3768..4559)
                     /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(3917..3936)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4730..5318
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5219..5238
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc
       61 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg
      121 ccaactccat cactaggggt tcctgcggcc gcacgcgtat caagtgccac ctgacgtctc
      181 cctatcagtg atagagaagt cgacacgtct cgagctccct atcagtgata gagaaggtac
      241 gtctagaacg tctccctatc agtgatagag aagtcgacac gtctcgagct ccctatcagt
      301 gatagagaag gtacgtctag aacgtctccc tatcagtgat agagaagtcg acacgtctcg
      361 agctccctat cagtgataga gaaggtacgt ctagaacgtc tccctatcag tgatagagaa
      421 gtcgacacgt ctcgagctcc ctatcagtga tagagaagct accccctata taagcagagc
      481 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
      541 aagacaccgg gaccgatcca gcctgattgt gctgtctcat cattttggca aagctagcct
      601 accggtcgcc accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
      661 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
      721 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
      781 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
      841 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
      901 caattgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
      961 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     1021 cgacggcccc gtaatgcagt gtcgtaccat gggctgggag gcctccactg agcggatgta
     1081 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     1141 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     1201 cgcctacaac gtcgacatca agttggacat cctttcccac aacgaggact acaccatcgt
     1261 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     1321 gtccggactc accggtggag gcggttcagg cggaggtggc tctggcggtg gcggatcgag
     1381 atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg gatccaccgg
     1441 atctagataa ctgatcaact cctcaggtgc aggctgccta tcaattcgat atcaagctta
     1501 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     1561 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     1621 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     1681 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     1741 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     1801 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     1861 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     1921 tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     1981 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     2041 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     2101 accgagcgct gctcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct
     2161 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg
     2221 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg
     2281 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag
     2341 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat
     2401 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta
     2461 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc
     2521 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac
     2581 cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg
     2641 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     2701 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     2761 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     2821 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     2881 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     2941 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3001 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3061 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3121 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3181 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     3241 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     3301 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3361 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3421 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3481 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     3541 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3601 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3661 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3721 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3781 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3841 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3901 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3961 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4021 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4081 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4141 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4201 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4261 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4321 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4381 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4441 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4501 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4561 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4621 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4681 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4741 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4801 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4861 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4921 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4981 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5041 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5101 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5161 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5221 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5281 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5341 tacggttcct ggccttttgc tggcc
//
