LOCUS       sequence-477364-        8660 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for transfection of cells for expression of C-FLAG PDZD8 
            (91-1154aa).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8660)
  AUTHORS   Li M, Wang Y, Wei X, Cai WF, Wu J, Zhu M, Wang Y, Liu YH, Xiong J, 
            Qu Q, Chen Y, Tian X, Yao L, Xie R, Li X, Chen S, Huang X, Zhang C, 
            Xie C, Wu Y, Xu Z, Zhang B, Jiang B, Wang ZC, Li Q, Li G, Lin SY, Yu
            L, Piao HL, Deng X, Han J, Zhang CS, Lin SC
  TITLE     AMPK targets PDZD8 to trigger carbon source shift from glucose to 
            glutamine.
  JOURNAL   Cell Res. 2024 Jun 19. doi: 10.1038/s41422-024-00985-6.
  PUBMED    38898113
REFERENCE   2  (bases 1 to 8660)
  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"; journalName: "Cell Res. 
            2024 Jun 19. doi: 10.1038/s41422-024-00985-6."
COMMENT     Sequence Label: pcDNA 3.3 C-FLAG PDZD8(91-1154aa)
FEATURES             Location/Qualifiers
     source          1..8660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        50..429
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        430..633
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     584..604
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     630..654
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3976..3999
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4046..4065)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4090..4138
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4340..4768
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4427..4446)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4637..4658
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4777..4797)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4782..5111
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4962..5097
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5024..5043
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5178..5972
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5232..5251)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5842..5861
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6148..6269
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6185..6204)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6239..6258
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6318..6334)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6318..6334)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6331..6353)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6342..6358
                     /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)."
     promoter        complement(join(6366..6372,6373..6390,6391..6396))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6411..6432
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6549..6566)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6720..7308)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6800..6819)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7479..8270,8271..8339))
                     /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     8102..8121
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8340..8444)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8519..8538)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actgagctcc ttaaggttaa ctctagagaa ttcggatcca tgacgcggga gacttgctac
      781 ttcctcaacg ccaccatcct attcctgttc cgggagttgc gggacaccgc gctgacccgc
      841 cgctgggtca ccaagaagat caaggtggag ttcgaggagc tgctgcagac caagacggcc
      901 gggcgcctgc tggaggggct gagcctgcgg gacgtgttcc tgggcgagac ggtgcccttc
      961 atcaagacca tccggctcgt gcggccagtc gtgccctcgg ccaccgggga gcccgatggc
     1021 cctgaagggg aggcgctgcc cgccgcctgc cccgaggagc tggccttcga ggcggaggtg
     1081 gagtacaacg ggggcttcca cctggccatc gacgtggacc tggtcttcgg caagtccgcc
     1141 tacttgtttg tcaagctgtc ccgcgtggtg ggaaggctgc gcttggtctt tacgcgcgtg
     1201 cccttcaccc actggttctt ctccttcgtg gaagacccgc tgatcgactt cgaggtgcgc
     1261 tcccagtttg aagggcggcc catgccccag ctcacctcca tcatcgtcaa ccagctcaag
     1321 aagatcatca agcgcaagca caccctaccg aattacaaga tcaggtttaa gccgtttttt
     1381 ccataccaga ccttgcaagg atttgaagaa gatgaagagc atatccatat acaacaatgg
     1441 gcacttactg aaggccgtct taaagttacg ttgttagaat gtagcaggtt actcattttt
     1501 ggatcctatg acagagaggc aaatgttcat tgtacacttg agttaagcag tagtgtttgg
     1561 gaagaaaaac agaggagttc tattaagacg gttgaattaa taaaaggaaa tttacaaagt
     1621 gttggactta cacttcgtct tgtccagtca actgatgggt atgctgggca cgtcatcatt
     1681 gaaactgtgg ctccaaactc gcctgctgca attgcagatc ttcagcgggg agatcgactt
     1741 atcgccattg gaggtgtgaa aatcacatca acactgcaag tgttgaagct tatcaagcag
     1801 gctggtgacc gagtcctggt gtactatgaa aggcctgttg gccagagtaa tcaaggtgca
     1861 gtgctgcaag ataactttgg ccagttggaa gaaaactttt tgtcaagctc atgccaatcg
     1921 ggttatgaag aggaagctgc cgggttgaca gtagatactg aaagtagaga gctggattct
     1981 gaatttgaag acttggcaag tgatgtcaga gcacaaaatg agttcaaaga tgaggcacaa
     2041 tcattaagtc atagtcccaa acgtgttcca acaacacttt ctattaaacc ccttggagct
     2101 atatcaccag ttttaaaccg taaattagct gtaggaagtc acccactacc accgaaaatt
     2161 cagtccaaag atggaaataa acctccaccc ctaaaaactt ctgagataac agacccagca
     2221 caagtgtcaa aaccaaccca aggatctgct ttcaaaccac ctgtgccacc acgaccacaa
     2281 gcgaaagttc ctttgccttc cgccgatgct ccaaatcagg cagaaccaga tgttctcgtt
     2341 gaaaagccag agaaggtggt gccacctcct cttgtagata aatctgctga aaagcaagca
     2401 aaaaatgtgg atgccataga cgatgcagct gcacctaagc aatttttagc aaagcaagaa
     2461 gtggccaaag atgtcacttc agaaacttcc tgccctacta aggacagttc ggacgaccgt
     2521 caaacatggg aatcatcaga aattctttat cgtaataagc taggaaaatg gacaagaacc
     2581 agagcatcct gtttgtttga catagaagcc tgtcacaggt acttaaacat tgcattgtgg
     2641 tgcagggatc ctttcaagtt gggaggtctc atctgtttgg ggcatgttag tttaaaactt
     2701 gaagatgtgg ctttaggatg cctagctaca tcaaacacgg aatacctttc caaattgaga
     2761 ctggaagccc cctcacctaa ggctatagtc actagaaccg cactacgcaa tctgagtatg
     2821 caaaagggat tcaatgacaa attttgctat ggtgacatta ctattcactt caaatatttg
     2881 aaagaaggag aatcagacca ccatgtagtt actaacgtag aaaaagaaaa agaaccccat
     2941 ttggttgaag aagtttctgt tctccctaaa gaggagcaat ttgttggaca gatgggttta
     3001 acagaaaaca aacacagttt tcaggatact cagttccaga acccaacatg gtgtgactac
     3061 tgtaagaaaa aagtttggac taaagcagct tcccagtgta tgttttgtgc ttatgtttgc
     3121 cataaaaaat gtcaagaaaa gtgtctagct gagacttctg tttgtggagc aactgatagg
     3181 cgaatagaca ggacactgaa aaaccttagg ctggaaggac aggaaaccct cttaggcctg
     3241 cctcctcgtg ttgatgctga agctagcaag tcagtcaata aaacaacagg tttgacaagg
     3301 catattatca atactagttc tcgtttatta aatttgcgtc aagtctctaa aactcgcctt
     3361 tctgaaccag gaaccgatct cgtagaacct tcaccaaaac acacacccaa cacgtcagac
     3421 aacgaaggca gtgacacgga ggtctgtggt ccaaacagtc cttctaaacg gggaaacagc
     3481 acaggaataa agttagtgag aaaagagggt ggtctggatg acagtgtttt cattgcagtt
     3541 aaagaaattg gtcgtgatct gtacaggggc ttgcctacag aggaaaggat ccagaaacta
     3601 gagttcatgt tggataagct acagaatgaa attgatcagg agttggaaca caataattcc
     3661 cttgttagag aagaaaaaga gacaactgat acaaggaaaa aatcacttct ttctgctgcc
     3721 ttagctaaat caggtgaaag gctacaagct ctaacacttc ttatgattca ctacagagca
     3781 ggcattgaag atatagaaac tttagaaagt ctgtctttag accagcactc caaaaaaata
     3841 agcaagtaca cagatgatac agaagaagac cttgataatg aaataagcca actaatagac
     3901 tctcagccat tcagcagcat atcagatgac ttatttggcc catccgagtc tgtgctcgag
     3961 ccatggaagc ttatggacta caaagacgat gacgacaagg atatctaact gactgaaccg
     4021 gtggtaccaa acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4081 gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat
     4141 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     4201 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     4261 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg
     4321 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4381 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4441 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4501 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4561 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4621 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4681 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4741 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     4801 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4861 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4921 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4981 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5041 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5101 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     5161 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     5221 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     5281 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     5341 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     5401 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     5461 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     5521 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5581 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5641 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5701 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5761 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5821 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5881 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5941 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc
     6001 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     6061 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     6121 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     6181 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6241 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     6301 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     6361 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6421 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6481 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6541 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6601 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6661 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6721 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6781 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6841 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6901 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6961 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7021 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7081 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7141 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7201 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7261 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7321 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7381 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7441 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7501 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7561 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7621 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7681 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7741 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7801 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7861 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7921 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7981 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8041 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     8101 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8161 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8221 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8281 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8341 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8401 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8461 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     8521 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     8581 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     8641 gcatgaagaa tctgcttagg
//
