LOCUS       sequence-477877-        8740 bp    DNA     circular SYN 08-JUL-2025
DEFINITION  Used for transfection of cells for expression of C-FLAG PDZD8 
            (66-1154aa).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8740)
  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 8740)
  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(66-1154aa)
FEATURES             Location/Qualifiers
     source          1..8740
                     /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             4051..4080
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4126..4145)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4170..4218
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4420..4848
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4507..4526)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4717..4738
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4857..4877)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4862..5191
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5042..5177
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5104..5123
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5258..6052
                     /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(5312..5331)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5922..5941
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6228..6349
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6265..6284)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6319..6338
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6398..6414)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6398..6414)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6411..6433)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6422..6438
                     /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(6446..6452,6453..6470,6471..6476))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6491..6512
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6629..6646)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6800..7388)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6880..6899)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7559..8350,8351..8419))
                     /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     8182..8201
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8420..8524)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8599..8618)
                     /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 tggatgagga gccctccgga
      781 gcggcccctg agggcggcgc gacccccacc gcggcccccg agacccccgc cccgccgacg
      841 cgggagactt gctacttcct caacgccacc atcctattcc tgttccggga gttgcgggac
      901 accgcgctga cccgccgctg ggtcaccaag aagatcaagg tggagttcga ggagctgctg
      961 cagaccaaga cggccgggcg cctgctggag gggctgagcc tgcgggacgt gttcctgggc
     1021 gagacggtgc ccttcatcaa gaccatccgg ctcgtgcggc cagtcgtgcc ctcggccacc
     1081 ggggagcccg atggccctga aggggaggcg ctgcccgccg cctgccccga ggagctggcc
     1141 ttcgaggcgg aggtggagta caacgggggc ttccacctgg ccatcgacgt ggacctggtc
     1201 ttcggcaagt ccgcctactt gtttgtcaag ctgtcccgcg tggtgggaag gctgcgcttg
     1261 gtctttacgc gcgtgccctt cacccactgg ttcttctcct tcgtggaaga cccgctgatc
     1321 gacttcgagg tgcgctccca gtttgaaggg cggcccatgc cccagctcac ctccatcatc
     1381 gtcaaccagc tcaagaagat catcaagcgc aagcacaccc taccgaatta caagatcagg
     1441 tttaagccgt tttttccata ccagaccttg caaggatttg aagaagatga agagcatatc
     1501 catatacaac aatgggcact tactgaaggc cgtcttaaag ttacgttgtt agaatgtagc
     1561 aggttactca tttttggatc ctatgacaga gaggcaaatg ttcattgtac acttgagtta
     1621 agcagtagtg tttgggaaga aaaacagagg agttctatta agacggttga attaataaaa
     1681 ggaaatttac aaagtgttgg acttacactt cgtcttgtcc agtcaactga tgggtatgct
     1741 gggcacgtca tcattgaaac tgtggctcca aactcgcctg ctgcaattgc agatcttcag
     1801 cggggagatc gacttatcgc cattggaggt gtgaaaatca catcaacact gcaagtgttg
     1861 aagcttatca agcaggctgg tgaccgagtc ctggtgtact atgaaaggcc tgttggccag
     1921 agtaatcaag gtgcagtgct gcaagataac tttggccagt tggaagaaaa ctttttgtca
     1981 agctcatgcc aatcgggtta tgaagaggaa gctgccgggt tgacagtaga tactgaaagt
     2041 agagagctgg attctgaatt tgaagacttg gcaagtgatg tcagagcaca aaatgagttc
     2101 aaagatgagg cacaatcatt aagtcatagt cccaaacgtg ttccaacaac actttctatt
     2161 aaaccccttg gagctatatc accagtttta aaccgtaaat tagctgtagg aagtcaccca
     2221 ctaccaccga aaattcagtc caaagatgga aataaacctc cacccctaaa aacttctgag
     2281 ataacagacc cagcacaagt gtcaaaacca acccaaggat ctgctttcaa accacctgtg
     2341 ccaccacgac cacaagcgaa agttcctttg ccttccgccg atgctccaaa tcaggcagaa
     2401 ccagatgttc tcgttgaaaa gccagagaag gtggtgccac ctcctcttgt agataaatct
     2461 gctgaaaagc aagcaaaaaa tgtggatgcc atagacgatg cagctgcacc taagcaattt
     2521 ttagcaaagc aagaagtggc caaagatgtc acttcagaaa cttcctgccc tactaaggac
     2581 agttcggacg accgtcaaac atgggaatca tcagaaattc tttatcgtaa taagctagga
     2641 aaatggacaa gaaccagagc atcctgtttg tttgacatag aagcctgtca caggtactta
     2701 aacattgcat tgtggtgcag ggatcctttc aagttgggag gtctcatctg tttggggcat
     2761 gttagtttaa aacttgaaga tgtggcttta ggatgcctag ctacatcaaa cacggaatac
     2821 ctttccaaat tgagactgga agccccctca cctaaggcta tagtcactag aaccgcacta
     2881 cgcaatctga gtatgcaaaa gggattcaat gacaaatttt gctatggtga cattactatt
     2941 cacttcaaat atttgaaaga aggagaatca gaccaccatg tagttactaa cgtagaaaaa
     3001 gaaaaagaac cccatttggt tgaagaagtt tctgttctcc ctaaagagga gcaatttgtt
     3061 ggacagatgg gtttaacaga aaacaaacac agttttcagg atactcagtt ccagaaccca
     3121 acatggtgtg actactgtaa gaaaaaagtt tggactaaag cagcttccca gtgtatgttt
     3181 tgtgcttatg tttgccataa aaaatgtcaa gaaaagtgtc tagctgagac ttctgtttgt
     3241 ggagcaactg ataggcgaat agacaggaca ctgaaaaacc ttaggctgga aggacaggaa
     3301 accctcttag gcctgcctcc tcgtgttgat gctgaagcta gcaagtcagt caataaaaca
     3361 acaggtttga caaggcatat tatcaatact agttctcgtt tattaaattt gcgtcaagtc
     3421 tctaaaactc gcctttctga accaggaacc gatctcgtag aaccttcacc aaaacacaca
     3481 cccaacacgt cagacaacga aggcagtgac acggaggtct gtggtccaaa cagtccttct
     3541 aaacggggaa acagcacagg aataaagtta gtgagaaaag agggtggtct ggatgacagt
     3601 gttttcattg cagttaaaga aattggtcgt gatctgtaca ggggcttgcc tacagaggaa
     3661 aggatccaga aactagagtt catgttggat aagctacaga atgaaattga tcaggagttg
     3721 gaacacaata attcccttgt tagagaagaa aaagagacaa ctgatacaag gaaaaaatca
     3781 cttctttctg ctgccttagc taaatcaggt gaaaggctac aagctctaac acttcttatg
     3841 attcactaca gagcaggcat tgaagatata gaaactttag aaagtctgtc tttagaccag
     3901 cactccaaaa aaataagcaa gtacacagat gatacagaag aagaccttga taatgaaata
     3961 agccaactaa tagactctca gccattcagc agcatatcag atgacttatt tggcccatcc
     4021 gagtctgtgc tcgagccatg gaagcttatg gaacaaaaac tcatctcaga agaggatctg
     4081 gatatctaac tgactgaacc ggtggtccaa acgggggagg ctaactgaaa cacggaagga
     4141 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     4201 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     4261 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     4321 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     4381 tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     4441 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4501 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4561 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4621 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4681 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4741 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4801 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4861 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4921 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4981 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     5041 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5101 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     5161 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     5221 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     5281 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     5341 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     5401 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     5461 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     5521 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     5581 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5641 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5701 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5761 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5821 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5881 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5941 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     6001 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     6061 ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     6121 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     6181 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     6241 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6301 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     6361 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     6421 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     6481 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     6541 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6601 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6661 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6721 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6781 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6841 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6901 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6961 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     7021 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7081 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7141 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7201 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     7261 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7321 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7381 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7441 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7501 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7561 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7621 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7681 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7741 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7801 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7861 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7921 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7981 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8041 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8101 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8161 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8221 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8281 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8341 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8401 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8461 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8521 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     8581 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     8641 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     8701 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
