LOCUS       sequence-412602-        5816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For AAV production to express C2m2-mKate in astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5816)
  AUTHORS   Dirvelyte E, Bujanauskiene D, Jankaityte E, Daugelaviciene N, 
            Kisieliute U, Nagula I, Budvytyte R, Neniskyte U
  TITLE     Genetically encoded phosphatidylserine biosensor for in vitro, ex 
            vivo and in vivo labelling.
  JOURNAL   Cell Mol Biol Lett. 2023 Jul 27;28(1):59. doi: 
            10.1186/s11658-023-00472-7.
  PUBMED    37501184
REFERENCE   2  (bases 1 to 5816)
  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: "Cell Mol 
            Biol Lett."; date: "2023-07-27"; volume: "28(1):59. doi"; pages: " 
            10.1186/s11658-023-00472-7"
COMMENT     Sequence Label: pAAV-C2m2-mKate
FEATURES             Location/Qualifiers
     source          1..5816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    836..854
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    872..890
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1616..2311
                     /codon_start=1
                     /product="monomeric red fluorescent protein optimized for 
                     use as a fusion tag (Shemiakina et al., 2012)"
                     /label=FusionRed
                     /note="mammalian codon-optimized"
                     /translation="VSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFMYGSKTFINHPQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPANGPVMQKKTLGWEASTETLYPADGGLEGACDMALK
                     LVGGGHLICNLETTYRSKKPAKNLKMPGVYNVDRRLERIKEADNETYVEQHEVAVARYS
                     TGGAGDGGK"
     misc_feature    2319..2907
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2372..2392)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2790..2801)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3470..3591
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3481..3500)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3535..3554
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3694..3823
                     /label=AAV2 ITR
     primer_bind     complement(3888..3906)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3974..4078
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4079..4147,4148..4939)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5110..5698
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5599..5618
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgcccgggc tttgcccggg
       61 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
      121 cctactagta acatatcctg gtgtggagta ggggacgctg ctctgacaga ggctcggggg
      181 cctgagctgg ctctgtgagc tggggaggag gcagacagcc aggccttgtc tgcaagcaga
      241 cctggcagca ttgggctggc cgccccccag ggcctcctct tcatgcccag tgaatgactc
      301 accttggcac agacacaatg ttcggggtgg gcacagtgcc tgcttcccgc cgcaccccag
      361 cccccctcaa atgccttccg agaagcccat tgagcagggg gcttgcattg caccccagcc
      421 tgacagcctg gcatcttggg ataaaagcag cacagccccc taggggctgc ccttgctgtg
      481 tggcgccacc ggcggtggag aacaaggctc tattcagcct gtgcccagga aaggggatca
      541 ggggatgccc aggcatggac agtgggtggc agggggggag aggagggctg tctgcttccc
      601 agaagtccaa ggacacaaat gggtgagggg agagctctcc ccatagctgg gctgcggccc
      661 aaccccaccc cctcaggcta tgccaggggg tgttgccagg ggcacccggg catcgccagt
      721 ctagcccact ccttcataaa gccctcgtat cccaggagcg agcagagcca gagcaggttg
      781 gagaggagac gcatcacctc tgctgcttgc ctaaaacagg taagtcccat taatctccct
      841 atcagtgata gagaaggtct gaagagttta ctccctatca gtgatagaga ttaattcctc
      901 tactaacctt gttcatcttt tctttttttt tctacaggtc ctgggtgatt aacagcttaa
      961 ggccgccacc atgcaggtct cccgtgtgct ggccgcgctg tgcggcatgc tactctgcgc
     1021 ctctggcctc ttcgccgcgt ctggtgacca tatgcacgga tgttctgagc ccctgggcct
     1081 gaagaataac acaattcctg acagccagat gtcagcctcc agcagctaca atacatggaa
     1141 cctgcgtgct tttggctggt acccccactt gggaaggctg gataatcagg gcaatatcaa
     1201 tgcctggacg gctcagagca acagtgccaa ggaatggctg caggttgacc tgggcactca
     1261 gaggcaagtg acaggaatca tcacccaggg ggcccgtgac tttggccaca tccagtatgt
     1321 ggcgtcctac aaggtagccc acagtgatga tggtgtgcag tggactgtat atgaggagca
     1381 aggaagcagc aaggtcttcc agggcaactt ggacaacaac tcccacaaga agaacatctt
     1441 cgagaaaccc ttcatggctc gctacgtgcg tgtccttcca gtgtcctggc ataaccgcat
     1501 caccctgcgc ctggagctgc tgggctgtaa gcttggcact ggaggctctg gaggcactgg
     1561 aggctctgga ggcaccggta gcactagcgg cggaagcggc gggacaggta cgcgtgtgag
     1621 cgagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tgaacaacca
     1681 ccacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag
     1741 aatcaaggtc gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctaccagctt
     1801 catgtacggc agcaaaacct tcatcaacca ccctcagggc atccccgact tctttaagca
     1861 gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct
     1921 gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg tcaagattag
     1981 aggggtgaac ttcccagcca acggccctgt gatgcagaag aaaacactcg gctgggaggc
     2041 ctccaccgag acgctgtacc ccgctgacgg cggcctggaa ggcgcatgtg acatggccct
     2101 gaagctcgtg ggcgggggcc acctgatctg caacttggag accacataca gatccaagaa
     2161 acccgctaag aacctcaaga tgcccggcgt ctacaacgtg gacaggagac tggaaagaat
     2221 caaggaggcc gacaatgaga cctacgtcga gcagcacgag gtggctgtgg ccagatactc
     2281 tactggtggc gctggtgatg gaggtaaatg aggatcctaa tcaacctctg gattacaaaa
     2341 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     2401 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     2461 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     2521 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     2581 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     2641 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     2701 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     2761 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     2821 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     2881 ggatctccct ttgggccgcc tccccgcctg aattctgcat acttctttac attccagata
     2941 tctgcatcaa acaccattgt cacactccat cgcgatcacg cgagccgaac gaacaaaccc
     3001 aacaacatga aactacctag cgcttctatt cagcatgatg tctttcgtat acacaaattc
     3061 ggttctacag ggtaacctag gctgcatgtt taaacataca tacttcttta cattccagat
     3121 atctgcatca aacaccattg tcacactcca tcgcgatcac gcgagccgaa cgaacaaacc
     3181 caacaacatg aaactaccta gcgcttctat tcagcatgat gtctttcgta tacacaaatt
     3241 cggttctaca gggtaaccta ggctgcatgt ttaaacatac atacttcttt acattccaga
     3301 tatctgcatc aaacaccatt gtcacactcc atcgcgatca cgcgagccga acgaacaaac
     3361 ccaacaacat gaaactacct agcgcttcta ttcagcatga tgtctttcgt atacacaaat
     3421 tcggttctac agggtaacct aggctgcagg taccttcgag cagacatgat aagatacatt
     3481 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     3541 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     3601 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag
     3661 taaaacctct acaaatgtgg taaaatcaag cttaggaacc cctagtgatg gagttggcca
     3721 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3781 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaagc
     3841 tagcgggcga ttaaggaaag ggctagatca ttcttgaaga cgaaagggcc tcgtgatacg
     3901 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     3961 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4021 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4081 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4141 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4201 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4261 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4321 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4381 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4441 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4501 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4561 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4621 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4681 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4741 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4801 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4861 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4921 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4981 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5041 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5101 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5161 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5221 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5281 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5341 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5401 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5461 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5521 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5581 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5641 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5701 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgta
     5761 ataaacacac acacaccaac aaccgtggtt ggttgttgtg ttggtttatt ctcgag
//
