LOCUS       sequence-356891-        7280 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pAAV construct GluN1A714L (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: pRB131
FEATURES             Location/Qualifiers
     source          1..7280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   14..143
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             634..1344
                     /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(661..689)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1284..1304
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4176..4196)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4556..4696
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4556..4685
                     /label=AAV2 ITR
     rep_origin      4771..5226
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4858..4877)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5068..5089
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5243..5262)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5362..5384
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5422..5440)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5508..5612
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5613..5681,5682..6473)
                     /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(5831..5850)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6644..7232
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7133..7152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg
       61 tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc
      121 caactccatc actaggggtt cctgcggccg cagatgctgc acacttgtgg actaagtttg
      181 ttcgcatccc cttctccaac cccctcagta catcaccctg ggggaacagg gtccacttgc
      241 tcctgggccc acacagtcct gcagtattgt gtatataagg ccagggcaaa gaggagcagg
      301 ttttaaagtg aaaggcaggc aggtgttggg gaggcagtta ccggggcaac gggaacaggg
      361 cgtttcggag gtggttgcca tggggacctg gatgctgacg aaggctcgcg aggctgtgag
      421 cagccacagt gccctgctca gaagccccaa gctcgtcagt caagccggtt ctccgtttgc
      481 actcaggagc acgggcaggc gagtggcccc tagttctggg ggcagcgtga aaagacggta
      541 tcgataagct tgattcgagc tagcatgagc accatgcacc tgctgacatt cgccctgctt
      601 ttttcctgct ccttcgcccg cgccgccact aggagtaaag gagaagaact tttcactgga
      661 gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtcagt
      721 ggagagggtg aaggtgatgc aacatacgga aaacttaccc ttaaatttat ttgcactact
      781 ggaaaactac ctgttccttg gccaacactt gtcactactt taacttatgg tgttcaatgc
      841 ttttcaagat acccagatca tatgaaacgg catgactttt tcaagagtgc catgcccgaa
      901 ggttatgtac aggaaagaac tatatttttc aaagatgacg ggaactacaa gacacgtgct
      961 gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg tattgatttt
     1021 aaagaagatg gaaacattct tggacacaaa ttggaataca actataacga tcaccaggtg
     1081 tacatcatgg cagacaaaca aaagaatgga atcaaagcta acttcaaaat tagacacaac
     1141 attgaagatg gaggcgttca actagcagac cattatcaac aaaatactcc aattggcgat
     1201 gggcccgtcc ttttaccaga caaccattac ctgtttacaa cttctactct ttcgaaagat
     1261 cccaacgaaa agagagacca catggtcctt cttgagtttg taacagctgc tgggattaca
     1321 catggcatgg atgaactata caaagctagt tgcgacccca agatcgtcaa catcggcgcg
     1381 gtgctgagca cgcgcaagca tgaacagatg ttccgcgagg cagtaaacca ggccaataag
     1441 cgacacggct cttggaagat acagctcaac gccacttctg tcacccacaa gcccaacgcc
     1501 atacagatgg ccctgtcagt gtgtgaggac ctcatctcta gccaggtcta cgctatccta
     1561 gttagccacc cgcctactcc caacgaccac ttcactccca cccctgtctc ctacacagct
     1621 ggcttctaca gaatccctgt cctgggactg actacccgaa tgtccatcta ctctgacaag
     1681 agtatccacc tgagtttcct tcgcacggtg ccgccctact cccaccagtc cagcgtctgg
     1741 tttgagatga tgcgagtcta caactggaac cacatcatcc tgctggtcag cgacgaccac
     1801 gagggacggg cagcgcagaa gcgcttggag acgttgctgg aggaacggga gtccaaggca
     1861 gagaaggtgc tgcagtttga cccaggaacc aagaatgtga cggctctgct gatggaggcc
     1921 cgggaactgg aggcccgggt catcatcctt tctgcaagcg aggacgacgc tgccacagtg
     1981 taccgcgcag ccgcaatgct gaacatgacg ggctctgggt acgtgtggct ggtcggggaa
     2041 cgcgagatct ctgggaacgc cctgcgctac gctcctgatg gcatcatcgg acttcagctc
     2101 atcaatggca agaatgagtc agcccacatc agtgacgccg tgggcgtggt ggcacaggca
     2161 gttcacgaac tcctagagaa ggagaatatc actgacccac cgcggggttg cgtgggcaac
     2221 accaacatct ggaagacagg accattgttc aagagggtgc tgatgtcttc taagtatgcg
     2281 gacggagtga ctggccgtgt ggaattcaat gaggatgggg accgcaaatt cgctaattac
     2341 agcatcatga acctgcagaa ccgcaagctg gtgcaagtgg gcatctacaa tggtacccat
     2401 gtcatcccaa atgacaggaa gatcatctgg ccaggaggag agacagagaa acctcgagga
     2461 taccagatgt ccaccagact aaagatagtg acaatccacc aagagccctt cgtgtacgtc
     2521 aagcccacaa tgagtgatgg gacatgcaaa gaggagttca cagtcaatgg tgacccagtg
     2581 aagaaggtga tctgtacggg gcctaatgac acgtccccag gcagcccacg ccacacagtg
     2641 ccccagtgct gctatggctt ctgcatagac ctgctcatca agctggcgcg gaccatgaat
     2701 tttacctatg aggtgcacct ggtggcagat ggcaagtttg gcacacagga gcgggtaaac
     2761 aacagcaaca aaaaggagtg gaacggaatg atgggcgagc tactcagtgg ccaagcggac
     2821 atgattgtgg caccactgac catcaacaat gagcgtgcgc agtacataga gttctccaag
     2881 cccttcaagt accagggcct gaccattttg gtcaagaagg agattcccag gagcacactg
     2941 gactcattta tgcagccttt tcagagcaca ctgtggttgc tagtaggact gtcagttcat
     3001 gtggtggctg tgatgctgta cctgctggac cgcttcagtc cctttggccg attcaaggtg
     3061 aacagtgagg aggaggagga agatgcactg accctgtcct ctgccatgtg gttttcctgg
     3121 ggcgtcctgc tcaactccgg cattggggaa ggtgcccccc ggagtttctc tgcacgtatc
     3181 ctaggcatgg tgtgggctgg tttcgccatg atcatagtgg cttcctacac tgccaacttg
     3241 gcagctttcc tggtgctgga tcggcctgag gagcgcatca cgggcatcaa tgaccccagg
     3301 ctcagaaacc cctcagacaa gttcatctac gcaactgtaa agcagagctc cgtggacatc
     3361 tacttccgga ggcaggtgga gttgagtacc atgtaccggc acatggaaaa acacaattac
     3421 gagagcctgg ctgaggccat ccaggctgtg cgggacaaca agctgcacgc ctttatctgg
     3481 gactcggccg tgctggagtt tgaggcttca cagaagtgcg atctggtgac cacgggtgag
     3541 ctgttcttcc gctcaggctt tggcatcggc atgcgcaagg acagcccctg gaagcagaac
     3601 gtttccctgt ccatactcaa gtcccatgag aatggcttca tggaagatct ggataagaca
     3661 tgggttcggt atcaggaatg cgactcccgc agcaatgctc ctgcaaccct cacttttgag
     3721 aacatggcag gggtcttcat gctggtggct ggaggcatcg tagctgggat tttcctcatt
     3781 ttcattgaga tcgcctacaa gcgacacaag gatgcccgta ggaagcagat gcagctggct
     3841 tttgcagccg tgaacgtgtg gaggaagaac ctgcaggata gaaagagtgg tagagcagag
     3901 cccgacccta aaaagaaagc cacatttagg gctatcacct ccaccctggc ctccagcttc
     3961 aagagacgta ggtcctccaa agacacgagc accgggggtg gacgcggcgc tttgcaaaac
     4021 caaaaagaca cagtgctgcc gcgacgcgct attgagaggg aggagggcca gctgcagctg
     4081 tgttcccgtc atagggagag ctgagataat tccgctcgag ataatcaacc tctggattac
     4141 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     4201 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     4261 tccttgtata aatcctggtt agttcttgcc acggcggaac tcatcgccgc ctgccttgcc
     4321 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt tatttgtgaa
     4381 atttgtgatg ctattgcttt atttgtaacc atctagcttt atttgtgaaa tttgtgatgc
     4441 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     4501 tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagcg gccgcaggaa
     4561 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     4621 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     4681 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4741 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     4801 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     4861 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4921 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4981 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5041 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     5101 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     5161 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     5221 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     5281 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     5341 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     5401 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     5461 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5521 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5581 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5641 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5701 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5761 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5821 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5881 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5941 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6001 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     6061 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6121 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6181 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6241 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6301 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6361 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6421 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6481 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6541 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6601 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6661 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6721 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6781 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6841 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6901 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6961 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7021 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7081 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7141 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7201 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7261 tcctggcctt ttgctggcct
//
