LOCUS       sequence-476060-        7208 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  To express a SEP tagged GluR2(Q) from a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7208)
  TITLE     Ploski Lab Plasmids 2025
REFERENCE   2  (bases 1 to 7208)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pAAV CMV-SEP-GluR2(Q)-pA
FEATURES             Location/Qualifiers
     source          1..7208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(127..837)
                     /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(167..187)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     782..810
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     complement(939..958)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     complement(1112..1136)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(1133..1336)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1162..1182)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1337..1640
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     repeat_region   1664..1793
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(1855..2443)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1935..1954)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2614..3405,3406..3474))
                     /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     3237..3256
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3475..3579)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3647..3665
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3703..3725)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3825..3844
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3861..4316)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3998..4019)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4210..4229
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   4391..4531
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4402..4531
                     /label=AAV2 ITR
     primer_bind     4570..4589
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 attgtcgata tggggtgtca gtctgaactc cgaagtggaa aactgaacca tccctacccg
       61 aaatgcactg tattcttgat cagcgcccct tggaaatagc ccccctatct gtatgctgtt
      121 gctagctttg tatagttcat ccatgccatg tgtaatccca gcagctgtta caaactcaag
      181 aaggaccatg tggtctctct tttcgttggg atctttcgaa agagtagaag ttgtaaacag
      241 gtaatggttg tctggtaaaa ggacgggccc atcgccaatt ggagtatttt gttgataatg
      301 gtctgctagt tgaacgcctc catcttcaat gttgtgtcta attttgaagt tagctttgat
      361 tccattcttt tgtttgtctg ccatgatgta cacctggtga tcgttatagt tgtattccaa
      421 tttgtgtcca agaatgtttc catcttcttt aaaatcaata ccttttaact cgattctatt
      481 aacaagggta tcaccttcaa acttgacttc agcacgtgtc ttgtagttcc cgtcatcttt
      541 gaaaaatata gttctttcct gtacataacc ttcgggcatg gcactcttga aaaagtcatg
      601 ccgtttcata tgatctgggt atcttgaaaa gcattgaaca ccataagtta aagtagtgac
      661 aagtgttggc caaggaacag gtagttttcc agtagtgcaa ataaatttaa gggtaagttt
      721 tccgtatgtt gcatcacctt caccctctcc actgacagaa aatttgtgcc cattaacatc
      781 accatctaat tcaacaagaa ttgggacaac tccagtgaaa agttcttctc ctttactcct
      841 aggagaagag acaccaaaaa tcagtcccca taaaacagga gaaaggagga cagaaatatg
      901 cataatcttt tgcatggtgg cggcaactgg gagtggacac ctgtggagag aaaggcaaag
      961 tggatgtcag taagaccaat aggtgcctat cagaaacgca agagtcttct ctgtctcgac
     1021 aagcccagtt tctattggtc tccttaaacc tgtcttgtaa ccttcggtgt cttctatgga
     1081 ggtcaaaaca gcgtggatgg cgtctccagg cgatctgacg gttcactaaa cgagctctgc
     1141 ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt
     1201 tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa
     1261 tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact
     1321 gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     1381 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     1441 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     1501 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     1561 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     1621 ttaccgtaag ttatgtaacg cggaactcca cgcgtgcggc cgcaggaacc cctagtgatg
     1681 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     1741 gcccgacgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggacatgtg
     1801 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1861 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1921 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1981 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2041 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2101 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2161 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2221 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2281 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2341 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2401 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2461 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2521 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2581 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2641 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2701 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2761 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     2821 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2881 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     2941 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3001 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3061 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3121 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3181 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3241 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3301 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3361 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3421 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3481 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3541 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3601 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     3661 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     3721 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     3781 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     3841 tgtactgaga gtgcaccata aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa
     3901 tttttgttaa atcagctcat tttttaacca atagaccgaa atcggcaaaa tcccttataa
     3961 atcaaaagaa tagcccgaga tagagttgag tgttgttcca gtttggaaca agagtccact
     4021 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     4081 actacgtgaa ccatcaccca aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     4141 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     4201 gagaaaggaa gggaagaaag cgaaaggagc gggcgctaag gcgctggcaa gtgtagcggt
     4261 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtacta
     4321 tggttgcttt gacgtatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     4381 atcaggcgcc cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag
     4441 cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag
     4501 ggagtggcca actccatcac taggggttcc tgcggccgct cggtccagtg aaaaaaatgc
     4561 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     4621 ctcgagacta gtggatccgc atgcactgct ttgggaaatt ctggagtcat aacacgtcca
     4681 ttttctccag tacttcctgt aattgacagc cttgccttgc tcctcatgac atcactcaag
     4741 gtcatcccct aaattttaac actctcgatg ccatatacgt tgtaaccttc cttataagtt
     4801 gcaaaattct gggaattctg cgaggaagat gggttaatat tctgtggatt ctttgccacc
     4861 ttcattcgtt tcgcctcggc ccttgacttg taacagaact caatcaaagc caccagcatt
     4921 gccaaaccaa ggcccccgac aaggatgtag aatactccag caacgttgct cagactgagg
     4981 gcactggtct tttccttgga atcacctccc ccgctgccgc actctccttt gtcgtaccac
     5041 catttgtttt tcaatttgtc caacaggcct tgttcattca gttttagtac tgcgaggtta
     5101 accgcatttc ctaatgagga tcctttaggt gtggcgatgc cgtagccttt ggaatccaag
     5161 tttcctccca ctttcatggt gtcacaaggc ttcctctgct cgatgtactc gttcattgtg
     5221 gactccagca agtaggcata ctttcctttg gatttccgga ctctggctac tccttctgcg
     5281 gtagtcctca caaacacaga gggctctgca ctcctcatat aagtccacat tttatcaaac
     5341 actgcgattt tagatctcct gaaaaactct ttagtggagc cagagtctaa tgttccataa
     5401 gcaatttctg tttgcttaga cagatcctca gcactttcga tgggagacac catcctctct
     5461 acagtcagga aggcagctaa gttagccgtg taggaggaga ttatgatcag ggtaaagaac
     5521 caccacacac ctccaacaat gcgcccagag agggatcttg gcgaaatatc gcatccctgc
     5581 tgcataaagg cacccaagga aaaccagaga ctattaaaaa tcccaaattc attagttgat
     5641 tcactacttt gtgtttctct tccatcttca aattcctcag tgtgccactc gtaggggcta
     5701 aatctgctga ccaggaataa aactacactg accccaatgt aggcaaacac aatgcacatc
     5761 cagatctcat aggctaaagg atcaagaaag gaaaacactc ctggtttgga cttctgaggc
     5821 ttcttgatca tgatagagat tccaagactc atgaagggct tggagaagtc aatcacctct
     5881 tctctcacga gagtgatagt taatggagca attgcaatgt cagctttccc gtagacaagc
     5941 tctccaacca taccattcca aattttggtg tcggcatccc tggccccata cttgccatcc
     6001 ccaacaatag tcagcttgta cttgaaccca cagtgtttgg caatttctgc agctaagtca
     6061 acacagtagc cctcgtaacg ctcattccct tcaagcattt catgattttt cttcatcata
     6121 acatatggag attccaatat tgtggtgacc accacagttt tgttttcaag cccagacgtg
     6181 tcatttcctg atgggagctc agttagggtg acaaccattt tatccacttc actccagtac
     6241 ccaatcttcc ggggtccatt tgttttgagc tccatgatgt taattgtgta gtttattcgt
     6301 tttccattct ggtcaaactt tatatttcca gagaggcctt caacttgaac ctgcttgagg
     6361 gccctttcta tttcgacccc ttgtccccag ggcacagctg ggttggccaa acaatcccct
     6421 gcatttcctc tccgggatat ttcaatcctc tgcttccgaa ggttacggaa tgcttcagtc
     6481 atcacttgga cagcatcata cgtcagggcc gaagtatact taattgtcgc tgtgtgtgct
     6541 ccagggtatt ctttctcttc cagtgttgac catctttcta taaatttaga caccagggaa
     6601 tcgtcgtagt ctacaatctg aaatccagag acatttgctc ctccaaactg aattttcagc
     6661 aggtccccat cagtgaatcc cagatttgca atgatataat ggtacccttt aacatgtttt
     6721 ccaatggtaa taacctggtc cacaatgtca tttactttat ccctttcaca gtccaggatt
     6781 acacgccgtt ctttttttaa ctccagatct tgaaagagcg atctgtaggt ctcatctttc
     6841 ttgtcattgt tgatgttccc cacattgata gcagtcacct gccacttctt ctctgcagca
     6901 gaatccagaa cagcttgcag tgttgataag cctctgtcac tgtcatagag gtatgcgaac
     6961 ttgtcccatt ggtagtactc aatcaagcta aggagtgctc ctttgaggtc aggtcgcatc
     7021 tggatgacaa atggatgtgt gccatctgtt gggaagctag gtgtgatgaa ggacacatgg
     7081 agtgtcccac agaatgatgt gatggtattt acagacttct tgtcataaaa tccaaaaatt
     7141 gcgtagactc ctcttgaaaa ctgggagcag aaagcattgg tgactgcgaa actgttggct
     7201 acctccaa
//
