LOCUS       sequence-426779-        7260 bp    DNA     circular SYN 14-APR-2025
DEFINITION  AAV packaging plasmid for GRAB PGD2-1.1 fluorescent sensor under 
            EF1alpha promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7260)
  AUTHORS   Sang D, Lin K, Yang Y, Ran G, Li B, Chen C, Li Q, Ma Y, Lu L, Cui 
            XY, Liu Z, Lv SQ, Luo M, Liu Q, Li Y, Zhang EE
  TITLE     Prolonged sleep deprivation induces a cytokine-storm-like syndrome 
            in mammals.
  JOURNAL   Cell. 2023 Dec 7;186(25):5500-5516.e21. doi: 
            10.1016/j.cell.2023.10.025. Epub 2023 Nov 27.
  PUBMED    38016470
REFERENCE   2  (bases 1 to 7260)
  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"; doi: 
            "10.1016/j.cell.2023.10.025"; journalName: "Cell"; date: "2023-12-7-
            7"; volume: "186"; issue: "25"; pages: "5500-5516.e21"
COMMENT     Sequence Label: pAAV-EF1a-PGD2-1.1
FEATURES             Location/Qualifiers
     source          1..7260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        101..1279
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          332..1270
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1227..1247
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     sig_peptide     1304..1366
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     protein_bind    1367..1391
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     2383..2404
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2510..2531)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2771..2790)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3257..3845
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3310..3330)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3728..3739)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3877..4353
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4076..4095)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4393..4533
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4393..4522
                     /label=AAV2 ITR
     rep_origin      4608..5063
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4695..4714)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4905..4926
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5080..5099)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5199..5221
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5259..5277)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5345..5449
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5450..5518,5519..6310)
                     /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(5668..5687)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6481..7069
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6970..6989
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   7131..7260
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga ggaatctttg
       61 cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg
      121 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
      181 cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
      241 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
      301 ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc
      361 tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct ggctgcagta
      421 cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc
      481 ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg
      541 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat
      601 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc
      661 gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg
      721 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg
      781 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat
      841 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg
      901 gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg
      961 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg
     1021 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag
     1081 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg
     1141 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct
     1201 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct
     1261 tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gacatggaga cagacacact
     1321 cctgctatgg gtactgctgc tctgggttcc aggttccact ggtgacacaa gtttgtacaa
     1381 aaaagttggc accaccggta agcagatgtt ctaccgctgc caaaacacca cctcggtgga
     1441 gaaagatcac tcggcgacga tgggctgggt gctcttcagc gccggcctcc tgggcaacct
     1501 gctggcgctg gggctgctgg cgcgctcggg gctggggtcg tgcccgctgc gctcgccacg
     1561 ccaggcgcct tcggtcttct gtgtgcttgt gtgtggcctg accatcaccg acttgcttgg
     1621 caagtgcctc ttgagccccg tcgtgctagc agcctacgcg cagaaccgga gcctgtgggg
     1681 actggtgtcc acgccgggca gctctctgtg ccaagccttc gcctttagta tgtctttctt
     1741 cggcctcgcc tcgacgctgc agctcctggc catggccctg gagtgttggc tatccctggg
     1801 acaccccttc ttctaccgac agtacatcac cctccgccgg ggcgtgctgg tggcgcccgt
     1861 cgtgagcacc ttctccttgg ctttctgcgc gctgcccttc gcaggcttcg gaaagttcgt
     1921 gcagtactgc cccggcactt ggtgcttcat ccagatggcc cacgaggagc gctcgccgtc
     1981 ggtgctgggc tactcagtgc tgtatgccag catcctggcg ctgctggtcc tcgccattgt
     2041 gctgtgcaac ctgagcgcca tgcgcaagct ctaccgcatg caccgccgat atcggagatg
     2101 gctcaacggc tggcctggcg agcccgcgcc ggccgggccg cgcgacaccg acgcgctgga
     2161 cctggaggag ggaggaaacg tctatatcaa ggccgacaag cagaagaacg gcatcaaggc
     2221 gaacttccag atccgccaca acatcgagga cggcggcgtg cagctcgcct accactacca
     2281 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcgt
     2341 gcagtccaaa ctttcgaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     2401 cgtgaccgcc gccgggatca ctgccggcat ggacgagctg tacaagggcg gtaccggagg
     2461 gagcatggtg agaaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2521 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggtgagg gcgatgccac
     2581 cgagggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2641 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     2701 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacatccagg agcgcaccat
     2761 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2821 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2881 gcacaagctg gagtacaaca ccggagcagc agcacgcatg aaggagcaca ttgaccacct
     2941 cattctcctg gctatcatga ccgtgctctt cgtgatgtgc tccctgcctt taacttatcg
     3001 tgcttactat ggagcattta aagctgtcac ccaggatgga acctctgaag aagtggaaga
     3061 ccttcgcgcc ttgcgtttta tatctgtgat ctcaattgtg gacccttgga ttttcatcat
     3121 tttcagaacc tctgtttttc ggatgttttt ccacaagatt ttcatcaggc ctctgaggta
     3181 tagaaattgg catagcaatc cctgccaaaa tgacatggaa tccagcctgt aagaattcga
     3241 tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     3301 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     3361 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     3421 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     3481 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     3541 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     3601 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     3661 ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta
     3721 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     3781 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     3841 cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg tgacccctcc
     3901 ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata
     3961 aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg
     4021 ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat
     4081 tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg
     4141 gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc
     4201 aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg
     4261 gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta
     4321 caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac cacgtgcgga
     4381 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     4441 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     4501 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     4561 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     4621 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4681 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4741 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4801 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     4861 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4921 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     4981 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     5041 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     5101 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     5161 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     5221 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     5281 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     5341 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5401 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     5461 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5521 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     5581 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5641 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5701 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5761 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5821 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5881 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5941 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6001 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6061 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6121 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     6181 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     6241 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6301 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     6361 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6421 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6481 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6541 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6601 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6661 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6721 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6781 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6841 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6901 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6961 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7021 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7081 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
     7141 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
     7201 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
//
