LOCUS       sequence-255093-        7011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid encoding mouse ACY1 with C-terminal flag tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7011)
  AUTHORS   Kim JT, Li VL, Terrell SM, Fischer CR, Long JZ
  TITLE     Family-wide Annotation of Enzymatic Pathways by Parallel In Vivo 
            Metabolomics.
  JOURNAL   Cell Chem Biol. 2019 Nov 21;26(11):1623-1629.e3. doi: 
            10.1016/j.chembiol.2019.09.009. Epub 2019 Oct 3.
  PUBMED    31587987
REFERENCE   2  (bases 1 to 7011)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.chembiol.2019.09.009"; journalName: "Cell Chem Biol"; 
            date: "2019-11-21- 21"; volume: "26"; issue: "11"; pages: 
            "1623-1629.e3"
COMMENT     Sequence Label: pENN.AAV.CB7.CI.ACY1-flag.WPRE.rBG
FEATURES             Location/Qualifiers
     source          1..7011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     enhancer        204..583
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        585..862
                     /label=chicken beta-actin promoter
     intron          863..1879
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1803..1825
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1887..1906
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3180..3203
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    3234..3822
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3287..3307)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3705..3716)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3848..3867)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3913..3968
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   4061..4190
                     /label=AAV2 ITR
     primer_bind     complement(4209..4226)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4209..4225)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4218..4240)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4367..4822
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4454..4473)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4664..4685
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4848..4952
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4953..5021,5022..5813)
                     /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(5171..5190)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5984..6572
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6473..6492
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6860..6881
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6896..6901,6902..6919,6920..6926)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6934..6950
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6939..6961
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6958..6974
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6958..6974
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctaccag ggtaatgggg
      181 atcctctaga actatagcta gtcgacattg attattgact agttattaat agtaatcaat
      241 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      301 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      361 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      421 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      481 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      541 tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac
      601 gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat
      661 tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccaggcgg
      721 ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca
      781 gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa
      841 aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc
      901 cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag
      961 cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt
     1021 ttcttttctg tggctgcgtg aaagccttga ggggctccgg gagggccctt tgtgcggggg
     1081 gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc
     1141 gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt
     1201 gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa
     1261 caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt
     1321 cgggctgcaa ccccccctgc acccccctcc ccgagttgct gagcacggcc cggcttcggg
     1381 tgcggggctc cgtacggggc gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca
     1441 ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg gagggctcgg gggaggggcg
     1501 cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc attgcctttt
     1561 atggtaatcg tgcgagaggg cgcagggact tcctttgtcc caaatctgtg cggagccgaa
     1621 atctgggagg cgccgccgca ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg
     1681 caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctccc
     1741 tctccagcct cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc
     1801 ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc
     1861 cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt
     1921 tggcaaagaa ttctgcagat atcgccacca tgaccaccaa ggatcccgag tctgagcacc
     1981 cctccgtgac gctcttccgc cagtacctgc gcatctgcac tgtgcaaccc aatccagact
     2041 atggaggggc catcaccttc ctggaggaga gagcccgcca gctgggtctg agctgtcaga
     2101 aaatagaggt ggtgcctggc tttgtgatca ctgtgctgac ctggccaggc accaacccgt
     2161 cactcccctc catcttgcta aactcccaca cagatgtggt acctgtcttc aaggaacatt
     2221 ggcatcacga cccctttgag gccttcaagg attccgaggg ctacatctat gctaggggct
     2281 cccaggacat gaagagtgtc agcatccagt acctggaagc tgtgaggagg ctgaagagcg
     2341 agggccaccg attccctagg accatccaca tgacctttgt gcctgatgag gaggtcggag
     2401 gtcacaaggg gatggagttg tttgtgaaga ggcctgagtt ccaggctctg cgggcaggct
     2461 tcgctctgga cgagggcctg gccaacccca cggatgcctt tactgtcttt tatagtgagc
     2521 ggagtccttg gtgggtacgg gttaccagca ctgggaagcc aggccatgcc tcacgcttca
     2581 ttgaggatac agctgcagag aagttgcata aggttataag ctccatcttg gcattccggg
     2641 agaaggaaag gcagaggctg caggcgaacc ctcacctgaa ggaaggcgct gtgacttcgg
     2701 tgaatctgac caagcttgaa ggtggcgtgg cctataatgt ggtacctgcc actatgagtg
     2761 ccagctttga cttccgagtg gcaccagatg tagacatgaa ggccttcgag aagcagctgc
     2821 agagatggtg ccaggaagcg ggagaggggg tcacctttga gtttgctcag aagtttacag
     2881 agcctcgaat gacacccacg gatgattcag atccctggtg ggcagctttc agcggagcct
     2941 gcaaggcaat gaacctcact ctggagccag agatctttcc tgctgccact gacagccgct
     3001 atatccgagc ggtggggatc ccagctctgg gcttctcgcc catgaaccgc acacctgtcc
     3061 tgctacatga ccataatgaa cggctgcatg aggatatatt tctccgtggg gttgacatat
     3121 atacaggcct cctctctgca cttgctagtg tgcccaccct gcctagtgaa agcggaggag
     3181 actacaagga cgatgacgac aagtagtaat gactcgagcc taagcttatc gataatcaac
     3241 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     3301 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     3361 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     3421 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     3481 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     3541 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     3601 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     3661 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     3721 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     3781 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgatgag
     3841 gatccgatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc
     3901 tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
     3961 gtctctcact cggaagcaat tcgttgatct gaatttcgac cacccataat acccattacc
     4021 ctggtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag
     4081 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     4141 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ccttaattaa
     4201 cctaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     4261 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     4321 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     4381 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4441 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4501 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4561 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4621 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4681 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4741 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     4801 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4861 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4921 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4981 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5041 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5101 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5161 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5221 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5281 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5341 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5401 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5461 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5521 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5581 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5641 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5701 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5761 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5821 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5881 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5941 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6001 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6061 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6121 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6181 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6241 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6301 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6361 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6421 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6481 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6541 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6601 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6661 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     6721 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     6781 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     6841 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     6901 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     6961 gaaacagcta tgaccatgat tacgccagat ttaattaagg ccttaattag g
//
