LOCUS       sequence-490175-        5841 bp    DNA     circular SYN 22-OCT-2025
DEFINITION  Mammalian expression vector for CitA-Tq-LR (low-range).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5841)
  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 5841)
  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-LR
FEATURES             Location/Qualifiers
     source          1..5841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        258..535
                     /label=chicken beta-actin promoter
     primer_bind     814..836
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     898..917
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(1001..1022)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1262..1281)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1835..2083
                     /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    2102..2690
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2155..2175)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2573..2584)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2733..2854
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2744..2763)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2798..2817
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3016..3145
                     /label=AAV2 ITR
     rep_origin      complement(3382..3970)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3462..3481)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4141..4932,4933..5001))
                     /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     4764..4783
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5002..5106)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5132..5587)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5269..5290)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5481..5500
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5659..5788
                     /label=AAV2 ITR
ORIGIN
        1 aagagtacca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
       61 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      121 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      181 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      241 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc
      301 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg
      361 ggcggggggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg
      421 ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt
      481 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt
      541 cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg
      601 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
      661 ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct
      721 tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggctgt ccgcgggggg
      781 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
      841 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
      901 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attggatcca ccatgggagt
      961 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1021 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1081 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1141 gaccaccctg tcctggggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
     1201 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1261 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1321 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1381 ggagtacaac ttcaccggcc ccaaagtgga cattacagaa gaacgcctgc attatcaagt
     1441 tggtcagcgc gcactgatcc aggctatgca gatctctgcc atgccagagc tggttgaggc
     1501 tgttcagaaa cgcgacctgg cccgcattaa ggccctgatc gaccctatgc gcagcttcag
     1561 cgacgccact tacatcactg ttggcgacgc ctccggccag cgtctctacc acgttaaccc
     1621 tgacgagatc ggcaaatcga tggagggtgg tgacagcgac gaagctttga tcaacgcggc
     1681 atcctacgtt agcgttcgca agggatccct gggcagctcc ctgcgcggca agtcccctat
     1741 ccaggacgcc accggcaagg ttatcggcat cgttagcgtt ggctacacca tcgaacagct
     1801 ggtggacaac gatttcaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
     1861 caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca
     1921 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1981 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tactggagtt
     2041 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg taccagtgaa agcttataga
     2101 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2161 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2221 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     2281 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     2341 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     2401 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     2461 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     2521 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     2581 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     2641 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     2701 tcgacccggg cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     2761 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2821 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2881 gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     2941 tggtaaaatc gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt
     3001 taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     3061 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     3121 cctcagtgag cgagcgagcg cgcagctgca ttaatgaatc ggccaacgcg cggggagagg
     3181 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3241 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3301 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3361 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3421 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3481 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3541 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3601 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3661 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3721 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3781 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3841 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3901 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3961 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4021 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4081 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4141 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4201 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4261 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4321 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4381 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4441 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4501 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4561 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4621 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4681 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4741 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4801 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4861 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4921 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4981 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5041 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5101 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     5161 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     5221 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     5281 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     5341 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     5401 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     5461 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     5521 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     5581 ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     5641 tcttcgctat tacgccagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
     5701 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
     5761 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
     5821 tctacgtagc catgctctag g
//
