LOCUS       sequence-479450-        6413 bp    DNA     circular SYN 23-JUL-2025
DEFINITION  Used for transfection of cells for expression of C-FLAG PDZD8 
            (840-1154aa).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6413)
  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 6413)
  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(840-1154aa)
FEATURES             Location/Qualifiers
     source          1..6413
                     /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             2782..2805
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2852..2871)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2896..2944
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3146..3574
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3233..3252)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3443..3464
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3583..3603)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3588..3917
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3768..3903
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3830..3849
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3984..4778
                     /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(4038..4057)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4648..4667
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4954..5075
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4991..5010)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5045..5064
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5124..5140)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5124..5140)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5137..5159)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5148..5164
                     /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(5172..5178,5179..5196,5197..5202))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5217..5238
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5355..5372)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5526..6114)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5606..5625)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6285..6413,1..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 ccatgaaaca cagttttcag gatactcagt tccagaaccc aacatggtgt
     1861 gactactgta agaaaaaagt ttggactaaa gcagcttccc agtgtatgtt ttgtgcttat
     1921 gtttgccata aaaaatgtca agaaaagtgt ctagctgaga cttctgtttg tggagcaact
     1981 gataggcgaa tagacaggac actgaaaaac cttaggctgg aaggacagga aaccctctta
     2041 ggcctgcctc ctcgtgttga tgctgaagct agcaagtcag tcaataaaac aacaggtttg
     2101 acaaggcata ttatcaatac tagttctcgt ttattaaatt tgcgtcaagt ctctaaaact
     2161 cgcctttctg aaccaggaac cgatctcgta gaaccttcac caaaacacac acccaacacg
     2221 tcagacaacg aaggcagtga cacggaggtc tgtggtccaa acagtccttc taaacgggga
     2281 aacagcacag gaataaagtt agtgagaaaa gagggtggtc tggatgacag tgttttcatt
     2341 gcagttaaag aaattggtcg tgatctgtac aggggcttgc ctacagagga aaggatccag
     2401 aaactagagt tcatgttgga taagctacag aatgaaattg atcaggagtt ggaacacaat
     2461 aattcccttg ttagagaaga aaaagagaca actgatacaa ggaaaaaatc acttctttct
     2521 gctgccttag ctaaatcagg tgaaaggcta caagctctaa cacttcttat gattcactac
     2581 agagcaggca ttgaagatat agaaacttta gaaagtctgt ctttagacca gcactccaaa
     2641 aaaataagca agtacacaga tgatacagaa gaagaccttg ataatgaaat aagccaacta
     2701 atagactctc agccattcag cagcatatca gatgacttat ttggcccatc cgagtctgtg
     2761 ctcgagccat ggaagcttat ggactacaaa gacgatgacg acaaggatat ctaactgact
     2821 gaaccggtgg taccaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     2881 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt
     2941 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     3001 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     3061 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc
     3121 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3181 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3241 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3301 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3361 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3421 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3481 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3541 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3601 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3661 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3721 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3781 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3841 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3901 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     3961 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4021 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4081 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4141 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4201 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4261 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4321 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4381 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4441 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4501 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4561 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4621 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4681 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4741 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa
     4801 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4861 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4921 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     4981 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5041 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     5101 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5161 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5221 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5281 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5341 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5401 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5461 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5521 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5581 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5641 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5701 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5761 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5821 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5881 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5941 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6001 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6061 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6121 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6181 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6241 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6301 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6361 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgc
//
