LOCUS       sequence-477902-        8048 bp    DNA     circular SYN 08-JUL-2025
DEFINITION  Used for transfection of cells for expression of C-FLAG PDZD8 
            (295-1154aa).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8048)
  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 8048)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 8, 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(295-1154aa)
FEATURES             Location/Qualifiers
     source          1..8048
                     /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             3364..3387
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3434..3453)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3478..3526
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3728..4156
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3815..3834)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4025..4046
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4165..4185)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4170..4499
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4350..4485
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4412..4431
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4566..5360
                     /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(4620..4639)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5230..5249
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5536..5657
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5573..5592)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5627..5646
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5706..5722)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5706..5722)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5719..5741)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5730..5746
                     /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(5754..5760,5761..5778,5779..5784))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5799..5820
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5937..5954)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6108..6696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6188..6207)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6867..7658,7659..7727))
                     /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     7490..7509
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7728..7832)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7907..7926)
                     /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 tgttttttcc ataccagacc
      781 ttgcaaggat ttgaagaaga tgaagagcat atccatatac aacaatgggc acttactgaa
      841 ggccgtctta aagttacgtt gttagaatgt agcaggttac tcatttttgg atcctatgac
      901 agagaggcaa atgttcattg tacacttgag ttaagcagta gtgtttggga agaaaaacag
      961 aggagttcta ttaagacggt tgaattaata aaaggaaatt tacaaagtgt tggacttaca
     1021 cttcgtcttg tccagtcaac tgatgggtat gctgggcacg tcatcattga aactgtggct
     1081 ccaaactcgc ctgctgcaat tgcagatctt cagcggggag atcgacttat cgccattgga
     1141 ggtgtgaaaa tcacatcaac actgcaagtg ttgaagctta tcaagcaggc tggtgaccga
     1201 gtcctggtgt actatgaaag gcctgttggc cagagtaatc aaggtgcagt gctgcaagat
     1261 aactttggcc agttggaaga aaactttttg tcaagctcat gccaatcggg ttatgaagag
     1321 gaagctgccg ggttgacagt agatactgaa agtagagagc tggattctga atttgaagac
     1381 ttggcaagtg atgtcagagc acaaaatgag ttcaaagatg aggcacaatc attaagtcat
     1441 agtcccaaac gtgttccaac aacactttct attaaacccc ttggagctat atcaccagtt
     1501 ttaaaccgta aattagctgt aggaagtcac ccactaccac cgaaaattca gtccaaagat
     1561 ggaaataaac ctccacccct aaaaacttct gagataacag acccagcaca agtgtcaaaa
     1621 ccaacccaag gatctgcttt caaaccacct gtgccaccac gaccacaagc gaaagttcct
     1681 ttgccttccg ccgatgctcc aaatcaggca gaaccagatg ttctcgttga aaagccagag
     1741 aaggtggtgc cacctcctct tgtagataaa tctgctgaaa agcaagcaaa aaatgtggat
     1801 gccatagacg atgcagctgc acctaagcaa tttttagcaa agcaagaagt ggccaaagat
     1861 gtcacttcag aaacttcctg ccctactaag gacagttcgg acgaccgtca aacatgggaa
     1921 tcatcagaaa ttctttatcg taataagcta ggaaaatgga caagaaccag agcatcctgt
     1981 ttgtttgaca tagaagcctg tcacaggtac ttaaacattg cattgtggtg cagggatcct
     2041 ttcaagttgg gaggtctcat ctgtttgggg catgttagtt taaaacttga agatgtggct
     2101 ttaggatgcc tagctacatc aaacacggaa tacctttcca aattgagact ggaagccccc
     2161 tcacctaagg ctatagtcac tagaaccgca ctacgcaatc tgagtatgca aaagggattc
     2221 aatgacaaat tttgctatgg tgacattact attcacttca aatatttgaa agaaggagaa
     2281 tcagaccacc atgtagttac taacgtagaa aaagaaaaag aaccccattt ggttgaagaa
     2341 gtttctgttc tccctaaaga ggagcaattt gttggacaga tgggtttaac agaaaacaaa
     2401 cacagttttc aggatactca gttccagaac ccaacatggt gtgactactg taagaaaaaa
     2461 gtttggacta aagcagcttc ccagtgtatg ttttgtgctt atgtttgcca taaaaaatgt
     2521 caagaaaagt gtctagctga gacttctgtt tgtggagcaa ctgataggcg aatagacagg
     2581 acactgaaaa accttaggct ggaaggacag gaaaccctct taggcctgcc tcctcgtgtt
     2641 gatgctgaag ctagcaagtc agtcaataaa acaacaggtt tgacaaggca tattatcaat
     2701 actagttctc gtttattaaa tttgcgtcaa gtctctaaaa ctcgcctttc tgaaccagga
     2761 accgatctcg tagaaccttc accaaaacac acacccaaca cgtcagacaa cgaaggcagt
     2821 gacacggagg tctgtggtcc aaacagtcct tctaaacggg gaaacagcac aggaataaag
     2881 ttagtgagaa aagagggtgg tctggatgac agtgttttca ttgcagttaa agaaattggt
     2941 cgtgatctgt acaggggctt gcctacagag gaaaggatcc agaaactaga gttcatgttg
     3001 gataagctac agaatgaaat tgatcaggag ttggaacaca ataattccct tgttagagaa
     3061 gaaaaagaga caactgatac aaggaaaaaa tcacttcttt ctgctgcctt agctaaatca
     3121 ggtgaaaggc tacaagctct aacacttctt atgattcact acagagcagg cattgaagat
     3181 atagaaactt tagaaagtct gtctttagac cagcactcca aaaaaataag caagtacaca
     3241 gatgatacag aagaagacct tgataatgaa ataagccaac taatagactc tcagccattc
     3301 agcagcatat cagatgactt atttggccca tccgagtctg tgctcgagcc atggaagctt
     3361 atggactaca aagacgatga cgacaaggat atctaactga ctgaaccggt ggtaccaaac
     3421 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     3481 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt
     3541 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     3601 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     3661 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg
     3721 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3781 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3841 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3901 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3961 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4021 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4081 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4141 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4201 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4261 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4321 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4381 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     4441 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4501 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     4561 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4621 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4681 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4741 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4801 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4861 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4921 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4981 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5041 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     5101 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5161 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5221 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5281 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5341 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     5401 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5461 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5521 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5581 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5641 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5701 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5761 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5821 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5881 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5941 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6001 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6061 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6121 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6181 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6241 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6301 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6361 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6421 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6481 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6541 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6601 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6661 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6721 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6781 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6841 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6901 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6961 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7021 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7081 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7141 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7201 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7261 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7321 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7381 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7441 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7501 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7561 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7621 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7681 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7741 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7801 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7861 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     7921 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     7981 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     8041 tgcttagg
//
