LOCUS       sequence-477898-        7832 bp    DNA     circular SYN 08-JUL-2025
DEFINITION  Used for transfection of cells for expression of C-FLAG PDZD8 
            (367-1154aa).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7832)
  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 7832)
  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(367-1154aa)
FEATURES             Location/Qualifiers
     source          1..7832
                     /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             3148..3171
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3218..3237)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3262..3310
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3512..3940
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3599..3618)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3809..3830
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3949..3969)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3954..4283
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4134..4269
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4196..4215
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4350..5144
                     /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(4404..4423)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5014..5033
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5320..5441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5357..5376)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5411..5430
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5490..5506)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5490..5506)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5503..5525)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5514..5530
                     /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(5538..5544,5545..5562,5563..5568))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5583..5604
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5721..5738)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5892..6480)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5972..5991)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6651..7442,7443..7511))
                     /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     7274..7293
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7512..7616)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7691..7710)
                     /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 tggttgaatt aataaaagga
      781 aatttacaaa gtgttggact tacacttcgt cttgtccagt caactgatgg gtatgctggg
      841 cacgtcatca ttgaaactgt ggctccaaac tcgcctgctg caattgcaga tcttcagcgg
      901 ggagatcgac ttatcgccat tggaggtgtg aaaatcacat caacactgca agtgttgaag
      961 cttatcaagc aggctggtga ccgagtcctg gtgtactatg aaaggcctgt tggccagagt
     1021 aatcaaggtg cagtgctgca agataacttt ggccagttgg aagaaaactt tttgtcaagc
     1081 tcatgccaat cgggttatga agaggaagct gccgggttga cagtagatac tgaaagtaga
     1141 gagctggatt ctgaatttga agacttggca agtgatgtca gagcacaaaa tgagttcaaa
     1201 gatgaggcac aatcattaag tcatagtccc aaacgtgttc caacaacact ttctattaaa
     1261 ccccttggag ctatatcacc agttttaaac cgtaaattag ctgtaggaag tcacccacta
     1321 ccaccgaaaa ttcagtccaa agatggaaat aaacctccac ccctaaaaac ttctgagata
     1381 acagacccag cacaagtgtc aaaaccaacc caaggatctg ctttcaaacc acctgtgcca
     1441 ccacgaccac aagcgaaagt tcctttgcct tccgccgatg ctccaaatca ggcagaacca
     1501 gatgttctcg ttgaaaagcc agagaaggtg gtgccacctc ctcttgtaga taaatctgct
     1561 gaaaagcaag caaaaaatgt ggatgccata gacgatgcag ctgcacctaa gcaattttta
     1621 gcaaagcaag aagtggccaa agatgtcact tcagaaactt cctgccctac taaggacagt
     1681 tcggacgacc gtcaaacatg ggaatcatca gaaattcttt atcgtaataa gctaggaaaa
     1741 tggacaagaa ccagagcatc ctgtttgttt gacatagaag cctgtcacag gtacttaaac
     1801 attgcattgt ggtgcaggga tcctttcaag ttgggaggtc tcatctgttt ggggcatgtt
     1861 agtttaaaac ttgaagatgt ggctttagga tgcctagcta catcaaacac ggaatacctt
     1921 tccaaattga gactggaagc cccctcacct aaggctatag tcactagaac cgcactacgc
     1981 aatctgagta tgcaaaaggg attcaatgac aaattttgct atggtgacat tactattcac
     2041 ttcaaatatt tgaaagaagg agaatcagac caccatgtag ttactaacgt agaaaaagaa
     2101 aaagaacccc atttggttga agaagtttct gttctcccta aagaggagca atttgttgga
     2161 cagatgggtt taacagaaaa caaacacagt tttcaggata ctcagttcca gaacccaaca
     2221 tggtgtgact actgtaagaa aaaagtttgg actaaagcag cttcccagtg tatgttttgt
     2281 gcttatgttt gccataaaaa atgtcaagaa aagtgtctag ctgagacttc tgtttgtgga
     2341 gcaactgata ggcgaataga caggacactg aaaaacctta ggctggaagg acaggaaacc
     2401 ctcttaggcc tgcctcctcg tgttgatgct gaagctagca agtcagtcaa taaaacaaca
     2461 ggtttgacaa ggcatattat caatactagt tctcgtttat taaatttgcg tcaagtctct
     2521 aaaactcgcc tttctgaacc aggaaccgat ctcgtagaac cttcaccaaa acacacaccc
     2581 aacacgtcag acaacgaagg cagtgacacg gaggtctgtg gtccaaacag tccttctaaa
     2641 cggggaaaca gcacaggaat aaagttagtg agaaaagagg gtggtctgga tgacagtgtt
     2701 ttcattgcag ttaaagaaat tggtcgtgat ctgtacaggg gcttgcctac agaggaaagg
     2761 atccagaaac tagagttcat gttggataag ctacagaatg aaattgatca ggagttggaa
     2821 cacaataatt cccttgttag agaagaaaaa gagacaactg atacaaggaa aaaatcactt
     2881 ctttctgctg ccttagctaa atcaggtgaa aggctacaag ctctaacact tcttatgatt
     2941 cactacagag caggcattga agatatagaa actttagaaa gtctgtcttt agaccagcac
     3001 tccaaaaaaa taagcaagta cacagatgat acagaagaag accttgataa tgaaataagc
     3061 caactaatag actctcagcc attcagcagc atatcagatg acttatttgg cccatccgag
     3121 tctgtgctcg agccatggaa gcttatggac tacaaagacg atgacgacaa ggatatctaa
     3181 ctgactgaac cggtggtacc aaacggggga ggctaactga aacacggaag gagacaatac
     3241 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg
     3301 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     3361 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     3421 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat
     3481 ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3541 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3601 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3661 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3721 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3781 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3841 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3901 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     3961 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4021 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4081 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4141 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4201 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4261 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4321 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4381 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4441 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4501 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4561 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4621 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4681 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4741 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4801 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4861 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4921 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4981 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5041 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5101 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5161 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     5221 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5281 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     5341 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5401 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     5461 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5521 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5581 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5641 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5701 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5761 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5821 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5881 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5941 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6001 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6061 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6121 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6181 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6241 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6301 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6361 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6421 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6481 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6541 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6601 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6661 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6721 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6781 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6841 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6901 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6961 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7021 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7081 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7141 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7201 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7261 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7321 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7381 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7441 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7501 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7561 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7621 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     7681 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     7741 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     7801 tgaccgacaa ttgcatgaag aatctgctta gg
//
