LOCUS       sequence-338898-        5441 bp    DNA     circular SYN 14-APR-2025
DEFINITION  AAV transfer plasmid for BREP (bioluminescent red protein).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5441)
  AUTHORS   Tian X, Zhang Y, Li X, Xiong Y, Wu T, Ai HW
  TITLE     A luciferase prosubstrate and a red bioluminescent calcium indicator
            for imaging neuronal activity in mice.
  JOURNAL   Nat Commun. 2022 Jul 8;13(1):3967. doi: 10.1038/s41467-022-31673-x.
  PUBMED    35803917
REFERENCE   2  (bases 1 to 5441)
  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: "Nat 
            Commun."; date: "2022-07-8"; volume: "13(1):3967. doi"; pages: " 
            10.1038/s41467-022-31673-x"
COMMENT     Sequence Label: pAAV-hSyn-BREP
FEATURES             Location/Qualifiers
     source          1..5441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   76..205
                     /label=AAV2 ITR
     promoter        281..728
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      757..766
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(896..914)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1164..1183
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1954..2542
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2007..2027)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2425..2436)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2545..2561)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2546..2562)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2591..2712
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2602..2621)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2656..2675
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2874..3003
                     /label=AAV2 ITR
     rep_origin      complement(3240..3828)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3320..3339)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3999..4790,4791..4859))
                     /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     4622..4641
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4860..4964)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(4990..5441,1..4))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5127..5148)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5339..5358
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
       61 tcgctattac gccagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg
      121 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
      181 gccaactcca tcactagggg ttccttgtag ttaatgatta acccgccatg ctacttatct
      241 acgtagccat gctctaggaa gatccagggc cctgcgtatg agtgcaagtg ggttttagga
      301 ccaggatgag gcggggtggg ggtgcctacc tgacgaccgg ccccgaccca ctggacaagc
      361 acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga
      421 tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc
      481 ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc
      541 ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg
      601 gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg
      661 ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga
      721 gagcgcagtc caattcaagc tgctagcaag gatcccgcca ccatggtgag caagggcgag
      781 gcagtgatca aggagttcat gcggttcaag gtgcacatgg agggctccat gaacggccac
      841 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
      901 ctgaaggtga ccaagggtgg ccccctgccc ttctcctggg acatcctgtc ccctcagttc
      961 atgtacggct ccagggcctt catcaagcac cccgccgaca tccccgacta ctataagcag
     1021 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgccgtg
     1081 accgtgaccc aggacacctc cctggaggac ggcaccctga tctacaaggt gaagctccgc
     1141 ggcaccaact tccctcctga cggccccgta atgcagaaga agacaatggg ctgggaagcg
     1201 tccaccgagc ggttgtaccc cgaggacggc gtgctgaagg gcgacattaa gatggccctg
     1261 cgcctgaagg acggcggccg ctacctggcg gacttcaaga ccacctacaa ggccaagaag
     1321 cccgtgcaga tgcccggcgc ctacaacgtc gaccgcaagt tggacatcac ctcccacaac
     1381 gaggactaca ccgtggtgga acagtacgaa cgctccgagg gccgccatct tgatacactc
     1441 gaagatttcg ttggggactg gcgacagaca gccggctaca acttgagtca agtccttgaa
     1501 cagggaggtg tgtccagttt gtttcagaat ctcggggtgt ccgtaactcc gatccaaagg
     1561 attgtcctga gcggtgaaaa tgggctgaag atcgacatcc atgtcatcat cccgtatgaa
     1621 ggtctgagcg gcgaccaaat gggccagatc gaaaaaattt ttaaggtggt gtaccctgtg
     1681 gataatcatc actttaaggt gatcctgcac tatggcacac tggtaatcga cggggttacg
     1741 ccgaacatga tcgactattt cggacggccg tatgaaggca tcgccgtgtt cgacggcaaa
     1801 aagatcactg taacagggac cctgtggaac ggcaacaaaa ttatcgacga gcgcctgatc
     1861 aaccccgacg gctccctgct gttccgagta accatcaacg gagtgaccgg ctggcgtctg
     1921 catgaacgca ttctggcgta aaagcttatc gataatcaac ctctggatta caaaatttgt
     1981 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     2041 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     2101 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     2161 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     2221 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     2281 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     2341 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     2401 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     2461 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     2521 tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gacccgggcg gccgcttcga
     2581 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     2641 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     2701 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg
     2761 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatctt
     2821 cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc
     2881 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     2941 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     3001 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3061 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3121 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3181 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3241 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3301 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3361 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3421 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3481 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3541 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3601 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3661 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3721 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3781 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3841 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3901 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3961 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4021 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4081 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4141 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4201 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4261 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4321 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4381 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4441 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4501 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4561 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4621 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4681 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4741 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4801 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4861 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4921 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4981 gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     5041 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     5101 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     5161 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     5221 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     5281 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     5341 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     5401 taaccaccac acccgccgcg cttaatgcgc cgctacaggg c
//
