LOCUS       sequence-490178-        5839 bp    DNA     circular SYN 22-OCT-2025
DEFINITION  Mammalian expression vector for CitA-Tq-HR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5839)
  AUTHORS   Rosen PC, Fu P, Ferran B, Kim E, Brooks DJ, Lim DC, Diaz-Garcia CM, 
            Yellen G
  TITLE     Activity-dependent citrate dynamics in neurons.
  JOURNAL   Proc Natl Acad Sci U S A. 2025 Oct 14;122(41):e2519902122. doi: 
            10.1073/pnas.2519902122. Epub 2025 Oct 10.
  PUBMED    41071660
REFERENCE   2  (bases 1 to 5839)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2025-10-14"; volume: "122(41):e2519902122. 
            doi"; pages: " 10.1073/pnas.2519902122. Epub 2025 Oct 10"
COMMENT     Sequence Label: pAAV-CAG-CitA-Tq-HR
FEATURES             Location/Qualifiers
     source          1..5839
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(213..1004,1005..1073))
                     /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     836..855
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1074..1178)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1204..1659)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1341..1362)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1553..1572
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1731..1860
                     /label=AAV2 ITR
     promoter        2171..2446
                     /label=chicken beta-actin promoter
     primer_bind     2725..2747
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2809..2828
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(2912..2933)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3173..3192)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             3746..3994
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELY"
     misc_feature    4013..4601
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4066..4086)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4484..4495)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4644..4765
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4655..4674)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4709..4728
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4927..5056
                     /label=AAV2 ITR
     rep_origin      complement(join(5293..5839,1..42))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5373..5392)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
       61 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      121 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      181 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
      241 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
      301 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
      361 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
      421 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
      481 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
      541 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
      601 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
      661 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
      721 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
      781 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
      841 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
      901 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
      961 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1021 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1081 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1141 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1201 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     1261 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     1321 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     1381 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     1441 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     1501 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     1561 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     1621 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     1681 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctgcgcgctc
     1741 gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg
     1801 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
     1861 tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct aggaagagta
     1921 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     1981 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     2041 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     2101 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     2161 attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
     2221 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
     2281 gggggggggg sgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg
     2341 gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag
     2401 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg
     2461 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
     2521 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta
     2581 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct
     2641 ccgggagggc cctttgtgcg gggggagcgg ctcggggctg tccgcggggg gacggctgcc
     2701 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag
     2761 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
     2821 ttattgtgct gtctcatcat tttggcaaag aattggatcc accatgggag tgagcaaggg
     2881 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2941 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     3001 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     3061 gtcctggggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     3121 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     3181 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     3241 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     3301 cttcaccggc cccaaagtgg acattacaga agaacgcctg cattatcaag ttggtcagcg
     3361 cgcactgatc caggctatgc agatctctgc catgccagag ctggttgagg ctgttcagaa
     3421 acgcgacctg gcccgcatta aggccctgat cgaccctatg cgcagcttca gcgacgccac
     3481 ttacatcact gttggcgacg cctccggcca gcgtctctac cacgttaacc ctgacgagat
     3541 cggcaaatcg atggagggtg gtgacagcga cgaagctttg atcaacgctg catcctacgt
     3601 tagcgttcgc aagggagccc tgggcagctc cctgcgcggc aagtccccta tccaggacgc
     3661 caccggcaag gttatcggca tcgttagcgt tggctacacc atcgaacagc tggtggacaa
     3721 cgatttcaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     3781 gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac
     3841 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa
     3901 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctactggagt tcgtgaccgc
     3961 cgccgggatc actctcggca tggacgagct gtaccagtga aagcttatcg ataatcaacc
     4021 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     4081 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     4141 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     4201 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     4261 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     4321 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     4381 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     4441 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     4501 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     4561 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacccgg
     4621 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     4681 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     4741 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     4801 tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat
     4861 cgataaggat cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta
     4921 actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     4981 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     5041 gcgagcgagc gcgcagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5101 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5161 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5221 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5281 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5341 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5401 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5461 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5521 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5581 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5641 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5701 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5761 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5821 tggtagcggt ggttttttt
//
