LOCUS       sequence-479448-        8927 bp    DNA     circular SYN 23-JUL-2025
DEFINITION  Used for transfection of cells for expression of C-FLAG PDZD8-WT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8927)
  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 8927)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 23, 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-WT
FEATURES             Location/Qualifiers
     source          1..8927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     495..514
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(733..837)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(912..931)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1103..1482
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1483..1686
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1637..1657
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1683..1707
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5296..5319
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5366..5385)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5410..5458
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5660..6088
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5747..5766)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5957..5978
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6097..6117)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6102..6431
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6282..6417
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6344..6363
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6498..7292
                     /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(6552..6571)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7162..7181
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7468..7589
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7505..7524)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7559..7578
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7638..7654)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7638..7654)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7651..7673)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7662..7678
                     /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(7686..7692,7693..7710,7711..7716))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7731..7752
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7869..7886)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8040..8628)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8120..8139)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8799..8927,1..663,664..732))
                     /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"
ORIGIN
        1 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
       61 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
      121 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
      181 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
      241 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
      301 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
      361 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
      421 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
      481 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
      541 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
      601 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
      661 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
      721 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
      781 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
      841 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
      901 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
      961 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     1021 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
     1081 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
     1141 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     1201 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     1261 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     1321 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     1381 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     1441 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     1501 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     1561 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     1621 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     1681 agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
     1741 catagaagac accgggaccg atccagcctc cggactgagc tccttaaggt taactctaga
     1801 gaattcggat ccatggggct gctgctcatg atcctggcgt cggccgtgct gggttccttc
     1861 ctcacgctcc tcgcccagtt cttcctgctg taccgcagac agcccgagcc gccggcggac
     1921 gaggccgccc gcgcgggcga gggcttccgc tacatcaagc cagtgccggg cctgctccta
     1981 agggagtacc tttatggcgg cggccgggat gaggagccct ccggagcggc ccctgagggc
     2041 ggcgcgaccc ccaccgcggc ccccgagacc cccgccccgc cgacgcggga gacttgctac
     2101 ttcctcaacg ccaccatcct attcctgttc cgggagttgc gggacaccgc gctgacccgc
     2161 cgctgggtca ccaagaagat caaggtggag ttcgaggagc tgctgcagac caagacggcc
     2221 gggcgcctgc tggaggggct gagcctgcgg gacgtgttcc tgggcgagac ggtgcccttc
     2281 atcaagacca tccggctcgt gcggccagtc gtgccctcgg ccaccgggga gcccgatggc
     2341 cctgaagggg aggcgctgcc cgccgcctgc cccgaggagc tggccttcga ggcggaggtg
     2401 gagtacaacg ggggcttcca cctggccatc gacgtggacc tggtcttcgg caagtccgcc
     2461 tacttgtttg tcaagctgtc ccgcgtggtg ggaaggctgc gcttggtctt tacgcgcgtg
     2521 cccttcaccc actggttctt ctccttcgtg gaagacccgc tgatcgactt cgaggtgcgc
     2581 tcccagtttg aagggcggcc catgccccag ctcacctcca tcatcgtcaa ccagctcaag
     2641 aagatcatca agcgcaagca caccctaccg aattacaaga tcaggtttaa gccgtttttt
     2701 ccataccaga ccttgcaagg atttgaagaa gatgaagagc atatccatat acaacaatgg
     2761 gcacttactg aaggccgtct taaagttacg ttgttagaat gtagcaggtt actcattttt
     2821 ggatcctatg acagagaggc aaatgttcat tgtacacttg agttaagcag tagtgtttgg
     2881 gaagaaaaac agaggagttc tattaagacg gttgaattaa taaaaggaaa tttacaaagt
     2941 gttggactta cacttcgtct tgtccagtca actgatgggt atgctgggca cgtcatcatt
     3001 gaaactgtgg ctccaaactc gcctgctgca attgcagatc ttcagcgggg agatcgactt
     3061 atcgccattg gaggtgtgaa aatcacatca acactgcaag tgttgaagct tatcaagcag
     3121 gctggtgacc gagtcctggt gtactatgaa aggcctgttg gccagagtaa tcaaggtgca
     3181 gtgctgcaag ataactttgg ccagttggaa gaaaactttt tgtcaagctc atgccaatcg
     3241 ggttatgaag aggaagctgc cgggttgaca gtagatactg aaagtagaga gctggattct
     3301 gaatttgaag acttggcaag tgatgtcaga gcacaaaatg agttcaaaga tgaggcacaa
     3361 tcattaagtc atagtcccaa acgtgttcca acaacacttt ctattaaacc ccttggagct
     3421 atatcaccag ttttaaaccg taaattagct gtaggaagtc acccactacc accgaaaatt
     3481 cagtccaaag atggaaataa acctccaccc ctaaaaactt ctgagataac agacccagca
     3541 caagtgtcaa aaccaaccca aggatctgct ttcaaaccac ctgtgccacc acgaccacaa
     3601 gcgaaagttc ctttgccttc cgccgatgct ccaaatcagg cagaaccaga tgttctcgtt
     3661 gaaaagccag agaaggtggt gccacctcct cttgtagata aatctgctga aaagcaagca
     3721 aaaaatgtgg atgccataga cgatgcagct gcacctaagc aatttttagc aaagcaagaa
     3781 gtggccaaag atgtcacttc agaaacttcc tgccctacta aggacagttc ggacgaccgt
     3841 caaacatggg aatcatcaga aattctttat cgtaataagc taggaaaatg gacaagaacc
     3901 agagcatcct gtttgtttga catagaagcc tgtcacaggt acttaaacat tgcattgtgg
     3961 tgcagggatc ctttcaagtt gggaggtctc atctgtttgg ggcatgttag tttaaaactt
     4021 gaagatgtgg ctttaggatg cctagctaca tcaaacacgg aatacctttc caaattgaga
     4081 ctggaagccc cctcacctaa ggctatagtc actagaaccg cactacgcaa tctgagtatg
     4141 caaaagggat tcaatgacaa attttgctat ggtgacatta ctattcactt caaatatttg
     4201 aaagaaggag aatcagacca ccatgtagtt actaacgtag aaaaagaaaa agaaccccat
     4261 ttggttgaag aagtttctgt tctccctaaa gaggagcaat ttgttggaca gatgggttta
     4321 acagaaaaca aacacagttt tcaggatact cagttccaga acccaacatg gtgtgactac
     4381 tgtaagaaaa aagtttggac taaagcagct tcccagtgta tgttttgtgc ttatgtttgc
     4441 cataaaaaat gtcaagaaaa gtgtctagct gagacttctg tttgtggagc aactgatagg
     4501 cgaatagaca ggacactgaa aaaccttagg ctggaaggac aggaaaccct cttaggcctg
     4561 cctcctcgtg ttgatgctga agctagcaag tcagtcaata aaacaacagg tttgacaagg
     4621 catattatca atactagttc tcgtttatta aatttgcgtc aagtctctaa aactcgcctt
     4681 tctgaaccag gaaccgatct cgtagaacct tcaccaaaac acacacccaa cacgtcagac
     4741 aacgaaggca gtgacacgga ggtctgtggt ccaaacagtc cttctaaacg gggaaacagc
     4801 acaggaataa agttagtgag aaaagagggt ggtctggatg acagtgtttt cattgcagtt
     4861 aaagaaattg gtcgtgatct gtacaggggc ttgcctacag aggaaaggat ccagaaacta
     4921 gagttcatgt tggataagct acagaatgaa attgatcagg agttggaaca caataattcc
     4981 cttgttagag aagaaaaaga gacaactgat acaaggaaaa aatcacttct ttctgctgcc
     5041 ttagctaaat caggtgaaag gctacaagct ctaacacttc ttatgattca ctacagagca
     5101 ggcattgaag atatagaaac tttagaaagt ctgtctttag accagcactc caaaaaaata
     5161 agcaagtaca cagatgatac agaagaagac cttgataatg aaataagcca actaatagac
     5221 tctcagccat tcagcagcat atcagatgac ttatttggcc catccgagtc tgtgctcgag
     5281 ccatggaagc ttatggacta caaagacgat gacgacaagg atatctaact gactgaaccg
     5341 gtggtaccaa acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     5401 gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat
     5461 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     5521 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     5581 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg
     5641 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     5701 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     5761 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     5821 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     5881 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5941 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     6001 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     6061 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     6121 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     6181 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     6241 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     6301 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     6361 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     6421 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     6481 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     6541 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     6601 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     6661 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     6721 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     6781 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     6841 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     6901 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     6961 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     7021 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     7081 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     7141 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     7201 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     7261 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc
     7321 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     7381 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     7441 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     7501 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     7561 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     7621 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     7681 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     7741 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     7801 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     7861 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     7921 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     7981 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     8041 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     8101 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     8161 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     8221 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     8281 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     8341 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     8401 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     8461 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     8521 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     8581 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     8641 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     8701 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     8761 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     8821 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     8881 tagataacta cgatacggga gggcttacca tctggcccca gtgctgc
//
