LOCUS       sequence-255096-        7074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid encoding mouse PM20D2 with C-terminal flag tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7074)
  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 7074)
  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.PM20D2-flag.WPRE.rBG
FEATURES             Location/Qualifiers
     source          1..7074
                     /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"
     regulatory      1944..1953
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3243..3266
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    3297..3885
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3350..3370)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3768..3779)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3911..3930)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3976..4031
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   4124..4253
                     /label=AAV2 ITR
     primer_bind     complement(4272..4289)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4272..4288)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4281..4303)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4430..4885
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4517..4536)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4727..4748
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4911..5015
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5016..5084,5085..5876)
                     /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(5234..5253)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6047..6635
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6536..6555
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6923..6944
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6959..6964,6965..6982,6983..6989)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6997..7013
                     /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     7002..7024
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7021..7037
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7021..7037
                     /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 tgggtccggt ggtggagcgg ccggccgagc
     1981 ccggcacgag cagcgccgcc gagctggagc tgctgaaacg gcgagcggcc gagcgcatcg
     2041 acgaggcggc cgagcgcctg ggcgcgctga gccgcgccat ctggagcgcg cccgagctgg
     2101 cctacgaaga gcaccgcgcg cacggcgagc tcacgcgctt cttcgagtgc gagccgccgg
     2161 ccgcctcgtg ggcggtgcag ccgcacttcg ggctccccac cgccttccgt gccgagtggg
     2221 cgccgccgga gtccgcggcg gggccccgcg cgctgcaggt ggccttcctg tgcgagtacg
     2281 acgcgctgcc agccctgggc cacgcctgcg gccacaacct catcgccgag gtcggggtgg
     2341 ctgcggcgct cggcctgcgg gctgcgctgg agagcatcgc tgcgccgccg ccggtcaagg
     2401 tgattgtcct gggaacccct gcagaagaag atggtggtgg caaaattgat ctaattgaag
     2461 caggtgcctt cgaaaacctt gatgttgttt ttatggctca tccctctcag gagaacgctg
     2521 cctatctccc ggatgtggcc gagcacgatg tgactgtgaa gtactacgga aaagcgtctc
     2581 acgctgccgc ttacccttgg gagggtgtga acgccttgga cgctgcagtt cttgcctata
     2641 ccaacctctc tgtgttaaga cagcagatga aaccaacctg gagagttcat ggtataataa
     2701 aaaacggtgg tgtgaagccc aatatcatcc cctcttactc tgaactcgtc tattacttcc
     2761 gtgcgccctc catgaaggag cttcaagttc tgaccaaaaa ggcagaagat tgcttcagag
     2821 ctgcggcttt ggctacgggc tgcacagtag acattgaaag tgaagcacat gattattaca
     2881 atgtcattcc caataagacc ctgtgcagtg cgtatacgga gaacgggaag aaactgggaa
     2941 tggagttcat ttcagaagat gcggtactga atggcccttc aggctctact gattttggaa
     3001 atgtcagttt cgtggttcct gggattcacc cgtattttta cattgggact gatgccttga
     3061 atcacacgga gcagtacaca gaagctgcag gatcacaagc ggctcagctg tacacgctgc
     3121 gtacagccaa ggctctggca atgaccgcac tggacgtcat ctttaagcct gcattgctcg
     3181 aaggagtcag gaaggagttt aaatgtaaac ttcaagaaga gcagctttta aatacagcag
     3241 cagactacaa ggacgatgac gacaagtagt aatgactcga gcctaagctt atcgataatc
     3301 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     3361 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     3421 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     3481 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     3541 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     3601 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     3661 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     3721 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     3781 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     3841 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgat
     3901 gaggatccga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
     3961 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     4021 tgtgtctctc actcggaagc aattcgttga tctgaatttc gaccacccat aatacccatt
     4081 accctggtag ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg
     4141 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     4201 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagccttaat
     4261 taacctaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     4321 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     4381 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     4441 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4501 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4561 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4621 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4681 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4741 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4801 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4861 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4921 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4981 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     5041 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     5101 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     5161 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     5221 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     5281 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     5341 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     5401 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     5461 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     5521 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     5581 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     5641 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     5701 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     5761 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5821 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     5881 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5941 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     6001 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6061 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6121 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6181 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6241 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6301 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6361 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6421 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6481 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6541 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6601 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6661 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6721 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     6781 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     6841 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     6901 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     6961 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     7021 caggaaacag ctatgaccat gattacgcca gatttaatta aggccttaat tagg
//
