LOCUS       sequence-356893-        7280 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pAAV construct GluN1WT (shRNA resistant).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7280)
  AUTHORS   Cappelli J, Khacho P, Wang B, Sokolovski A, Bakkar W, Raymond S, 
            Ahlskog N, Pitney J, Wu J, Chudalayandi P, Wong AYC, Bergeron R
  TITLE     Glycine-induced NMDA receptor internalization provides 
            neuroprotection and preserves vasculature following ischemic stroke.
  JOURNAL   iScience. 2021 Dec 3;25(1):103539. doi: 10.1016/j.isci.2021.103539. 
            eCollection 2022 Jan 21.
  PUBMED    34977503
REFERENCE   2  (bases 1 to 7280)
  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: "iScience.";
            date: "2021-12-3"; volume: "25(1):103539. doi"; pages: " 
            10.1016/j.isci.2021.103539. eCollection 2022 Jan 21"
COMMENT     Sequence Label: pRB112
FEATURES             Location/Qualifiers
     source          1..7280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   88..217
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             708..1418
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Ng et al., 2002)"
                     /label=superecliptic pHluorin
                     /note="excited strongly by blue light at neutral pH but 
                     minimally fluorescent at acidic pH"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(735..763)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1358..1378
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4250..4270)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4630..4770
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4630..4759
                     /label=AAV2 ITR
     rep_origin      4845..5300
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4932..4951)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5142..5163
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5317..5336)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5436..5458
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5496..5514)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5582..5686
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5687..5755,5756..6547)
                     /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(5905..5924)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6718..7280,1..26)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7207..7226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
       61 ccttttgctg gccttttgct cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg
      121 aggccgcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc
      181 agagagggag tggccaactc catcactagg ggttcctgcg gccgcagatg ctgcacactt
      241 gtggactaag tttgttcgca tccccttctc caaccccctc agtacatcac cctgggggaa
      301 cagggtccac ttgctcctgg gcccacacag tcctgcagta ttgtgtatat aaggccaggg
      361 caaagaggag caggttttaa agtgaaaggc aggcaggtgt tggggaggca gttaccgggg
      421 caacgggaac agggcgtttc ggaggtggtt gccatgggga cctggatgct gacgaaggct
      481 cgcgaggctg tgagcagcca cagtgccctg ctcagaagcc ccaagctcgt cagtcaagcc
      541 ggttctccgt ttgcactcag gagcacgggc aggcgagtgg cccctagttc tgggggcagc
      601 gtgaaaagac ggtatcgata agcttgattc gagctagcat gagcaccatg cacctgctga
      661 cattcgccct gcttttttcc tgctccttcg cccgcgccgc cactaggagt aaaggagaag
      721 aacttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca
      781 aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat
      841 ttatttgcac tactggaaaa ctacctgttc cttggccaac acttgtcact actttaactt
      901 atggtgttca atgcttttca agatacccag atcatatgaa acggcatgac tttttcaaga
      961 gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat gacgggaact
     1021 acaagacacg tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa
     1081 aaggtattga ttttaaagaa gatggaaaca ttcttggaca caaattggaa tacaactata
     1141 acgatcacca ggtgtacatc atggcagaca aacaaaagaa tggaatcaaa gctaacttca
     1201 aaattagaca caacattgaa gatggaggcg ttcaactagc agaccattat caacaaaata
     1261 ctccaattgg cgatgggccc gtccttttac cagacaacca ttacctgttt acaacttcta
     1321 ctctttcgaa agatcccaac gaaaagagag accacatggt ccttcttgag tttgtaacag
     1381 ctgctgggat tacacatggc atggatgaac tatacaaagc tagttgcgac cccaagatcg
     1441 tcaacatcgg cgcggtgctg agcacgcgca agcatgaaca gatgttccgc gaggcagtaa
     1501 accaggccaa taagcgacac ggctcttgga agatacagct caacgccact tctgtcaccc
     1561 acaagcccaa cgccatacag atggccctgt cagtgtgtga ggacctcatc tctagccagg
     1621 tctacgctat cctagttagc cacccgccta ctcccaacga ccacttcact cccacccctg
     1681 tctcctacac agctggcttc tacagaatcc ctgtcctggg actgactacc cgaatgtcca
     1741 tctactctga caagagtatc cacctgagtt tccttcgcac ggtgccgccc tactcccacc
     1801 agtccagcgt ctggtttgag atgatgcgag tctacaactg gaaccacatc atcctgctgg
     1861 tcagcgacga ccacgaggga cgggcagcgc agaagcgctt ggagacgttg ctggaggaac
     1921 gggagtccaa ggcagagaag gtgctgcagt ttgacccagg aaccaagaat gtgacggctc
     1981 tgctgatgga ggcccgggaa ctggaggccc gggtcatcat cctttctgca agcgaggacg
     2041 acgctgccac agtgtaccgc gcagccgcaa tgctgaacat gacgggctct gggtacgtgt
     2101 ggctggtcgg ggaacgcgag atctctggga acgccctgcg ctacgctcct gatggcatca
     2161 tcggacttca gctcatcaat ggcaagaatg agtcagccca catcagtgac gccgtgggcg
     2221 tggtggcaca ggcagttcac gaactcctag agaaggagaa tatcactgac ccaccgcggg
     2281 gttgcgtggg caacaccaac atctggaaga caggaccatt gttcaagagg gtgctgatgt
     2341 cttctaagta tgcggacgga gtgactggcc gtgtggaatt caatgaggat ggggaccgca
     2401 aattcgctaa ttacagcatc atgaacctgc agaaccgcaa gctggtgcaa gtgggcatct
     2461 acaatggtac ccatgtcatc ccaaatgaca ggaagatcat ctggccagga ggagagacag
     2521 agaaacctcg aggataccag atgtccacca gactaaagat agtgacaatc caccaagagc
     2581 ccttcgtgta cgtcaagccc acaatgagtg atgggacatg caaagaggag ttcacagtca
     2641 atggtgaccc agtgaagaag gtgatctgta cggggcctaa tgacacgtcc ccaggcagcc
     2701 cacgccacac agtgccccag tgctgctatg gcttctgcat agacctgctc atcaagctgg
     2761 cgcggaccat gaattttacc tatgaggtgc acctggtggc agatggcaag tttggcacac
     2821 aggagcgggt aaacaacagc aacaaaaagg agtggaacgg aatgatgggc gagctactca
     2881 gtggccaagc ggacatgatt gtggcaccac tgaccatcaa caatgagcgt gcgcagtaca
     2941 tagagttctc caagcccttc aagtaccagg gcctgaccat tttggtcaag aaggagattc
     3001 ccaggagcac actggactca tttatgcagc cttttcagag cacactgtgg ttgctagtag
     3061 gactgtcagt tcatgtggtg gctgtgatgc tgtacctgct ggaccgcttc agtccctttg
     3121 gccgattcaa ggtgaacagt gaggaggagg aggaagatgc actgaccctg tcctctgcca
     3181 tgtggttttc ctggggcgtc ctgctcaact ccggcattgg ggaaggtgcc ccccggagtt
     3241 tctctgcacg tatcctaggc atggtgtggg ctggtttcgc catgatcata gtggcttcct
     3301 acactgccaa cttggcagct ttcctggtgc tggatcggcc tgaggagcgc atcacgggca
     3361 tcaatgaccc caggctcaga aacccctcag acaagttcat ctacgcaact gtaaagcaga
     3421 gctccgtgga catctacttc cggaggcagg tggagttgag taccatgtac cggcacatgg
     3481 aaaaacacaa ttacgagagc gcagctgagg ccatccaggc tgtgcgggac aacaagctgc
     3541 acgcctttat ctgggactcg gccgtgctgg agtttgaggc ttcacagaag tgcgatctgg
     3601 tgaccacggg tgagctgttc ttccgctcag gctttggcat cggcatgcgc aaggacagcc
     3661 cctggaagca gaacgtttcc ctgtccatac tcaagtccca tgagaatggc ttcatggaag
     3721 atctggataa gacatgggtt cggtatcagg aatgcgactc ccgcagcaat gctcctgcaa
     3781 ccctcacttt tgagaacatg gcaggggtct tcatgctggt ggctggaggc atcgtagctg
     3841 ggattttcct cattttcatt gagatcgcct acaagcgaca caaggatgcc cgtaggaagc
     3901 agatgcagct ggcttttgca gccgtgaacg tgtggaggaa gaacctgcag gatagaaaga
     3961 gtggtagagc agagcccgac cctaaaaaga aagccacatt tagggctatc acctccaccc
     4021 tggcctccag cttcaagaga cgtaggtcct ccaaagacac gagcaccggg ggtggacgcg
     4081 gcgctttgca aaaccaaaaa gacacagtgc tgccgcgacg cgctattgag agggaggagg
     4141 gccagctgca gctgtgttcc cgtcataggg agagctgaga taattccgct cgagataatc
     4201 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     4261 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     4321 ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg
     4381 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     4441 tgtttatttg tgaaatttgt gatgctattg ctttatttgt aaccatctag ctttatttgt
     4501 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     4561 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     4621 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     4681 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     4741 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     4801 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     4861 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4921 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4981 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     5041 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     5101 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     5161 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     5221 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     5281 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     5341 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     5401 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     5461 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     5521 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     5581 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5641 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5701 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5761 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5821 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5881 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5941 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     6001 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6061 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     6121 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6181 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6241 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6301 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6361 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6421 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6481 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6541 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6601 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6661 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6721 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6781 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6841 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6901 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6961 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7021 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7081 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7141 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7201 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7261 gcgtcgattt ttgtgatgct
//
