LOCUS       sequence-385072-        8154 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of AMPA GluA1 with N terminal HaloTag fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8154)
  AUTHORS   Kim D, Park P, Li X, Wong Campos JD, Tian H, Moult EM, Grimm JB, 
            Lavis L, Cohen AE
  TITLE     Mapping memories: pulse-chase labeling reveals AMPA receptor 
            dynamics during memory formation.
  JOURNAL   bioRxiv. 2023 May 29:2023.05.26.541296. doi: 
            10.1101/2023.05.26.541296. Preprint.
  PUBMED    37292614
REFERENCE   2  (bases 1 to 8154)
  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: "bioRxiv. 
            2023 May 29:2023.05.26.541296. doi: 10.1101/2023.05.26.541296. 
            Preprint."
COMMENT     Sequence Label: TL028_AAV_hSyn:HaloTag-GluA1
FEATURES             Location/Qualifiers
     source          1..8154
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        167..614
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(652..685)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(736..769)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4493..4526
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    4577..4610
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4635..5223
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4688..4708)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5106..5117)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5277..5296)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5331..5350
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5409..5549
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5409..5538
                     /label=AAV2 ITR
     rep_origin      5624..6079
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5711..5730)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5921..5942
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6096..6115)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6215..6237
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6275..6293)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6361..6465
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6466..6534,6535..7326)
                     /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(6684..6703)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7497..8085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7986..8005
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(8147..8154,1..122)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc
       61 ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc
      121 ctgcggccgc acgcgtgtgt ctagactgca gagggccctg cgtatgagtg caagtgggtt
      181 ttaggaccag gatgaggcgg ggtgggggtg cctacctgac gaccgacccc gacccactgg
      241 acaagcaccc aacccccatt ccccaaattg cgcatcccct atcagagagg gggaggggaa
      301 acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca gtgccttcgc
      361 ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg acgtcactcg
      421 ccggtccccc gcaaactccc cttcccggcc accttggtcg cgtccgcgcc gccgccggcc
      481 cagccggacc gcaccacgcg aggcgcgaga taggggggca cgggcgcgac catctgcgct
      541 gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt
      601 gcctgagagc gcagtcgaga aggtaccgga tcctctagag tcgactccgg aataacttcg
      661 tataggatac tttatacgaa gttatgcaga atggtagctg gattgtagct gctattagca
      721 atatgaaacc tcttaataac ttcgtatagc atacattata cgaagttatg gcgcgccctt
      781 acaatcctgt ggctcccaag ggcatccctg aactgtgact catgcaggga atggattgca
      841 tggacttggg gaagtcctgg ctgaccaccc ggccattctc tccactgccg cctcctccgc
      901 tggctcctgc cccactgttc cgggggaggg tcgatgtccg tatggcttca ttgatggatt
      961 gctgtgggat caaacagaaa cccttcatcc gcttcgactc gctacgggat ttgtagcaga
     1021 actcgattaa ggcaaccagc atggccagtc ccagccctcc aatcaggatg tagaacacgc
     1081 ctgccacatt gctgaggctc aaagcgctgg tcttgtcctt ggagtcacct cccccgctgc
     1141 cgcactcgcc cttgtcgtac caccatttgt ttttcaattt gtccaaaagc ccctgctcgt
     1201 tcagttttaa cactgccagg tttactggat ttctcagggc ggaccccttg ggtgtcgcaa
     1261 tgccatagcc tttggaatcc aagttacctc ccactttcat ggtgtcacag ggctttcgtt
     1321 gctcaatata ctcattcatg gtggactcca ggaggtaggc gtatttgcct ttagattttc
     1381 tcactctgat catgccttcc tctgtggtcc gaacaaacac ggatggttct gcagacttca
     1441 tgtatgtcca catcttctca aacacagcga ttttagatct cctgaagaac tccttagtgg
     1501 agcctgcttc caatgtccca taagcaattt ccgtctgctt tgccaggtcc tctgcactct
     1561 caatgggaga caccatcctc tccacagtca ggaaggcagc caggttggct gtgtacgagg
     1621 agatgatgat caaagtgaag aaccaccaga cgccgccgac gatgcgtccg gacagggacc
     1681 tgggggaaat gtcacatcct tgctgcatga aggcccccag ggagaaccac aggctgttga
     1741 atatgccaaa ctcatttgac tggtcactgg ttgtctggtc tcgtccctct tcaaactctt
     1801 cgctgtgcca ttcgtagggg ctgaaacggc tgaccaggaa gaggacgacg ctcactccaa
     1861 tgtaggcaaa cactatacac atccagatct cataggccaa agggtcaaga aaggagaaga
     1921 cacctggctt ggacttctgt ggcttcttaa tcataatgga gattcccaaa ctcatgaatg
     1981 gcttggagaa gtcgatgact tcctcccgga ccaaggttat ggtcaaggga gccacagcca
     2041 cgtctgctct tccatagacc agttctccca ccatgccatt ccaagccttt gtgtcgggat
     2101 cccgggctcc atatttgccg tcgctgacaa tctcaagtcg gtaggagtag cccacgtgct
     2161 tggcgatctc tgcagccagc tccacacagt agccctcata gcggtcattg ccctcaaact
     2221 ggttggcatt ctttttaagc atcacgtaag gatcttcgag gatagtagtg acgatgtagg
     2281 tcctattctg gacgcttgag ttgtcccctc cagcctgagc gtcggtggct gcggggacaa
     2341 atttatcgtc ttcattccag taaccaatct ttcggattcc atcatgtttc atttcgatca
     2401 cgtggagggt gtaattggtc cggcgccctt tctcgttgaa ctgcacattt cctgtcaaac
     2461 cttcgaagcg cacctgctgc agggctctct ggatgtcgat cccttgaccc cagggcacag
     2521 ctgggttagc cagacagtcc ccagcattcc cccggcggga tatgtcaatc ctctgcctcc
     2581 gcaggctttg gaaggcctca gccatcacct tgacaccatc ataggtgaga gcagaagtgt
     2641 actttggcct cttccagtcc accctggtat ggtctcggga gtcacttgtc ctccattgct
     2701 gcatgattct ggctgggatc gtgtctgtgt agttcaccag ctggaaacct gtcacattgg
     2761 ctccgctctc cttgaactta tttaagtcaa tgtccatgaa gcccagattg gcgaggatgt
     2821 agtggtaccc gatgccattc ttttctagct tcacgatctg gcccaggatg gcgttgaggc
     2881 gttctgattc acagtcaacc accaccagcc tctccttttt cttctccagg tcctgaaaga
     2941 gcatccggta tccttcctcg gtggttgtca gaatgttgac agccgttacc tgccagttct
     3001 tctctgcggc tgtatccaag actctctgca ggactgacag gccccggtca gcatcataaa
     3061 tgtagacaaa ggtttgccac ttgtaatggt cgataatgct aatgagagct tcctgtagtt
     3121 ccgggcgtag ctgaaggaca aattgattgg atgtgtcaac aggaaaactt ggagtaatga
     3181 agcacacatg gagggcccca cagaaggagg tcagcatgtt gacagtcctt cgttcataaa
     3241 atccaaagat ggcatagact cctttggaga actgggaaca gaaacggtaa gtcatctcaa
     3301 agctgtcgct gatgttcaca atatcgatct ggggaagcag cttggggggc tccgtgagtt
     3361 gtgacaaagc aaacctaaaa gccgcatgtt cctgtgattg ttggtttgga aataaccccc
     3421 ctatctggat attgttgggg aagcgcgaac ccccgccacc tccgccccct ccacctccac
     3481 gcgtagtggt tggctcgccg gaaatctcga gcgtcgacag ccagcgcgcg atctcgctgc
     3541 cgatcaggtc cgggttgtct tcttgcagca gattcagacc cgggccgatg tccacagcct
     3601 tgcagttagg caggcttttg gccaggcgag cggcttcggc cggtgggatc agaacgcctg
     3661 gggtgcccca gaacagcagc ttcgggacag gggactggtg cagccagtcc atgtattctt
     3721 cgaccagcgc gacgatgttc gctggctcac cggcgattgg cagctcgttt gggaagcgcc
     3781 acagtggctc gcggtcaaca ggattcagga acggctcgcg gtaatggtcc atctcgactt
     3841 cagtcagcgg gcggacgaca cccatcggca gcgtaccctc gataaaaacg ttctgatcga
     3901 tgatcagctt gcggccgacg tcggtggtgc ggaaggcctg gaaggtctcg cgggcaaatt
     3961 ctggccattc gtcccaggtc gggatagggc ggatgaactc cataaatgca atacctttga
     4021 cgcgctctgg attgcgcttg gcccagtgga aacccagagc ggagccccag tcgtgaatga
     4081 ccaggacgac ctcttccaga cccagggctt cgatgaaggc atccatgaag cggacgtggt
     4141 cgtcgaagaa ataacccagg tctggtttgt cggatttgcc cataccgatc aggtctggag
     4201 caatgcagcg atgggtcggt gcaacatgcg ggatgatgtt gcgccacacg taggaggagg
     4261 tcgggttacc gtgcaggaac agcacagggg tgccatcgcg cggaccaaca tcgacgtagt
     4321 gcatgcgctc gcccaggact tccacataat gggggtcgaa tggaaagcca gtaccgatac
     4381 gcgttcctcc accccctccc ccaccgcctc cacctcgcgc attggcaccc acaaccgcac
     4441 ctagaaaacc ggtgcagaaa aaggcaaaga tgtacggcat ggtggcgcta gcataacttc
     4501 gtataaagta tcctatacga agttatttgc cttaacccag aaattatcac tgttattctt
     4561 tagaatggtg caaagaataa cttcgtataa tgtatgctat acgaagttat gaattcgata
     4621 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     4681 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     4741 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     4801 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     4861 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     4921 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     4981 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     5041 ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     5101 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     5161 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     5221 cgcatcgata ccgtcgaccc gggcggccgc ttcgagcaga catgataaga tacattgatg
     5281 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     5341 atgctattgc tttatttgta accattataa gctgcaataa actaaccacg tgcggaccga
     5401 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     5461 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     5521 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     5581 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     5641 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5701 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5761 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     5821 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5881 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5941 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     6001 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     6061 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     6121 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     6181 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     6241 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     6301 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     6361 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     6421 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     6481 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6541 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     6601 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     6661 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     6721 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6781 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6841 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6901 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6961 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     7021 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     7081 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     7141 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     7201 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     7261 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     7321 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     7381 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     7441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7621 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     7681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     8041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     8101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcctg cagg
//
