LOCUS       sequence-412604-        5663 bp    DNA     circular SYN 14-APR-2025
DEFINITION  For AAV production to express C2-SNAP in astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5663)
  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 5663)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-C2-SNAP
FEATURES             Location/Qualifiers
     source          1..5663
                     /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..2158
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="DKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVEV
                     PAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLKV
                     VKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLAV
                     KEWLLAHEGHRLGKPGLG"
     misc_feature    2166..2754
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2219..2239)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2637..2648)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3317..3438
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3328..3347)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3382..3401
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3541..3670
                     /label=AAV2 ITR
     primer_bind     complement(3735..3753)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3821..3925
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3926..3994,3995..4786)
                     /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      4957..5545
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5446..5465
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgcccgggc wwwgcccggg
       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 agacatggaa
     1141 cctgcgtgct tttggctggt acccccactt gggaaggctg gataatcagg gcaagatcaa
     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 cgcgtgacaa
     1621 agactgcgaa atgaagcgca ccaccctgga tagccctctg ggcaagctgg aactgtctgg
     1681 gtgcgaacag ggcctgcacc gtatcatctt cctgggcaaa ggaacatctg ccgccgacgc
     1741 cgtggaagtg cctgccccag ccgccgtgct gggcggacca gagccactga tgcaggccac
     1801 cgcctggctc aacgcctact ttcaccagcc tgaggccatc gaggagttcc ctgtgccagc
     1861 cctgcaccac ccagtgttcc agcaggagag ctttacccgc caggtgctgt ggaaactgct
     1921 gaaagtggtg aagttcggag aggtcatcag ctacagccac ctggccgccc tggccggcaa
     1981 tcccgccgcc accgccgccg tgaaaaccgc cctgagcgga aatcccgtgc ccattctgat
     2041 cccctgccac cgggtggtgc agggcgacct ggacgtgggg ggctacgagg gcgggctcgc
     2101 cgtgaaagag tggctgctgg cccacgaggg ccacagactg ggcaagcctg ggctgggtgg
     2161 atcctaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2221 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2281 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2341 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     2401 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     2461 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     2521 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     2581 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     2641 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     2701 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctgaat
     2761 tctgcatact tctttacatt ccagatatct gcatcaaaca ccattgtcac actccatcgc
     2821 gatcacgcga gccgaacgaa caaacccaac aacatgaaac tacctagcgc ttctattcag
     2881 catgatgtct ttcgtataca caaattcggt tctacagggt aacctaggct gcatgtttaa
     2941 acatacatac ttctttacat tccagatatc tgcatcaaac accattgtca cactccatcg
     3001 cgatcacgcg agccgaacga acaaacccaa caacatgaaa ctacctagcg cttctattca
     3061 gcatgatgtc tttcgtatac acaaattcgg ttctacaggg taacctaggc tgcatgttta
     3121 aacatacata cttctttaca ttccagatat ctgcatcaaa caccattgtc acactccatc
     3181 gcgatcacgc gagccgaacg aacaaaccca acaacatgaa actacctagc gcttctattc
     3241 agcatgatgt ctttcgtata cacaaattcg gttctacagg gtaacctagg ctgcaggtac
     3301 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     3361 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3421 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3481 gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcaagctt
     3541 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     3601 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     3661 gagcgcgcag agagggagtg gccaagctag cgggcgatta aggaaagggc tagatcattc
     3721 ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3781 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3841 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3901 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3961 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4021 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4081 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4141 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     4201 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4261 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4321 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4381 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4441 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4501 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4561 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4621 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4681 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4741 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4801 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4861 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4921 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4981 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5041 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5101 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5161 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5221 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5281 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5341 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5401 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5461 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5521 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5581 cttttgctgg ccttttgctc acatgtaata aacacacaca caccaacaac cgtggttggt
     5641 tgttgtgttg gtttattctc gag
//
