LOCUS       sequence-479443-       10282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for microinjection of nematodes for expression of KGA-33A, GLS 
            with 33 residues mutated to alanine at the interface for interacting
            with PDZD8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10282)
  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 10282)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 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: pHM1 KGA-33A
FEATURES             Location/Qualifiers
     source          1..10282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     183..200
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    317..338
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(353..358,359..376,377..383)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    391..407
                     /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)."
     primer_bind     396..418
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     415..431
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     415..431
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6555..6583)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     7331..7351
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     protein_bind    complement(7428..7452)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(8278..8300)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8400..8419
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(8501..8519)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8587..8691
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8692..8760,8761..9552)
                     /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     complement(8910..8929)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(9723..10282,1..29)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10212..10231
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
       61 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
      121 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
      181 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
      241 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
      301 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
      361 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
      421 acagctatga ccatgattac gccaagcttg catgcgcggc cgcactgact gggccggcct
      481 gctaaattgc aaccccgtgt ttttgaaaaa atacatcata acagagaaaa ctctttaaaa
      541 tcatattgtt cagccaggtc acagtcaagg acaattttga atttgactca atttcagtta
      601 caggctcgga taacggcgct gccacgacca ccctggcccc ttctgaccca acttgttaaa
      661 tattaatcta aatacaaata agatttagta tgaacattta ctgttcattt tgaatgttgt
      721 ttttttacac attttcgtag ttgaaaattt tgacatattt taagtgttgg aaaagatatg
      781 agacatttga gcgaactcgt ccaaatttgt gaaaatttcc aaaaatcatc gtgaaaaatt
      841 aaaaaacata ttttcagaga aaaggcttaa acatttttga atatgaataa gtaacctttt
      901 ttaaaaatcg gcatgttttt ttctttgaag atcaatattt ttgcgaaagc ctacgatact
      961 aaaaaatatt aacagcaaaa aggtgtaaaa ttaaaaagtg attgtttcat ctgttgacaa
     1021 aagttttgtc aaacaactgg ttttggaatt tatatgcaaa aacctgtaaa aagagtgttt
     1081 attgaatctt ggacttgact tgtttcatcg gactgtaatt tagcaggggc ttaagatagg
     1141 atgaattggt aaagtgataa aatatagcta ttggtatgca aaatattgtt acaattgatt
     1201 actttttggt atcattcacg tttctgaagt taccgccact caaagttaaa taaaataatt
     1261 tttgcaagca tttttgacaa cttaaacgaa tatggcggga ccaaaaaggc catgcctgga
     1321 gctgttattg catgtttttg ataaacttac tgtcatacat cagggtacat aaggtttttg
     1381 atttcacaca aagaaaaaaa aatcgaaaaa aaaacaattt ttttccgttg tcatagttga
     1441 aaaaccctgt tgctaaacaa ttcttctaat ttttcgtggt aactgctaca caatcggatt
     1501 aaggaaacct aattttttct cacatatgtt ggagagcctc gatattcttg aactttttgc
     1561 attcattgtt taatgtttct tcacatagct tcgtatatat tgccagcaat gcaaatcttg
     1621 agccatcggt tgaaatgatt ttgaaaaggt gctcggcata tctgggctga aaaaaaactt
     1681 ttgtttcaac gcattaaaca cttttcgggg ttctgttcca attttttaaa tagttgtaac
     1741 acccgctaga atttccgttt caccaaagaa atctttgttc acagattatt atatatttct
     1801 tttttatgta ttttgtccag gtgatcctaa atgtttaaaa aacgttccag cagcttttag
     1861 tctatacgaa gtatattttt tcttattttg gtttatcgga ggtatgcagt tccaaatgtt
     1921 ggattttcta ccctactaga aatttccttt agttatacaa ttttaattcg aagttgtatc
     1981 ttgaaaagga aaccatcatc aacaacaaat aagcttgaga ctagaattgc taatcgtctg
     2041 caccaaatat gaaattcgtt gaataaatta tataaaacag ttgacaatca taaagtattt
     2101 tacatacata tgtatagtta atgtcatatt agcatattta gtattgagtt aacacgcaga
     2161 aataccaaag aaaccctgtt taattcattt ccatgtttta ttcgctttaa ctagggaatg
     2221 accagaataa gtggagcgat attcaaaaaa aaaaattgta tcggaaagct gacattctct
     2281 actataagaa tatgactgaa atttttgccc attcgggctg gaaatctgaa tggaaattgt
     2341 ttatatgagg ggatccatgg ttgtagttac aattttgaga aaaaagttta aattttaggg
     2401 gaaaagatgt tgaatgaatc attatggtgt tggtcgacac attgatcatg ctgatatcga
     2461 tcagaaaaat taaatttggg cttttatata gtttcgaaaa aacacgttta aatttttgaa
     2521 aaattttacg ttagggtgga attgaacccc agagtcccac catgcttgtc atccactcta
     2581 tccactcggc catctcgctg ttgcaggcag tactaattgg ggatagtgaa aagagttaac
     2641 gtagaatttc agacgtaaaa atttcagatt tccagcccga atgggcaaaa atttcagtca
     2701 tattcttata gtagagaatg tcagctttcc gatacaatat tttttttttg aatatcgctc
     2761 cacttattct ggtcattccc tagtaagttt tttcagtttt accagaaata aaaccactca
     2821 taaaaactat aaactacaca tttaaactgt aaatgttgga aatcattttg accatcaaac
     2881 aatagctatc ggaaaatgca aaggccattc cgtgatggca cgctcttctt ctatccatcg
     2941 ctccttttca acgaaacaca cacacaaacg cagactattg gaaaagaaaa atatagaaaa
     3001 aacctttttt ctccttgtcc cttgtgttga tctcactttt cagtggacgt gtgcagtgtg
     3061 gaggccagag actgtgctga ccgacggacg cgcaggcggt aaacatacgt tgcagtggag
     3121 agtgtagtcc ctcgtcggct ctctgtgtat ttctttcaat gcctgtgtct cggtctatct
     3181 ttgtctctat ctgttttaaa agttttttgc aaagtttcaa tttctttttt attttctctt
     3241 ctttttttgc gcatttattt tttttaaaga gcgctgttct tcaattggaa aacagtataa
     3301 cgatcgaaat aaaaaaaaaa cttgaaatag tatcggaatt gagatggggg caattgaaat
     3361 tgaagacgca acaaaaacat gtttttatcg tctgaaatta tctaaacttt gaatctaacg
     3421 tgtcaatggc tacttttaca gtagactttt tagagtaatg ttaaaaagga taatataggt
     3481 ttttcttcaa actattttgt cacgccgcat atattctttt catattccat tcgcaaagtg
     3541 catgatgtaa caaaaatgaa acgtcataat tttttaatct ttcacacttt ttttgccggc
     3601 cgttaaatca aaatcaaatc aacttaatcc aacacgtgcg ggcttgcaat tttattacac
     3661 cggtgtccta cagcgccatg ttatcttctt tgattagaga tctctcacgc aacatttgac
     3721 attgaaaatt ttgtctccga attgttgtgt acctactttg tcccgatgtc ccaaattttc
     3781 actgataaca agagatatct caactgttct tatcacgttg atacgtagaa cgttttacac
     3841 taaaccaaat tgcagtcccc ttctacgaac cgtttgctaa ttgttttgca tagaacagtg
     3901 atcaatgtgt tgtgctcact agtcaatgat gttcgtctct catttttcct acaacttcac
     3961 tgaaaaggtt ttgaatatct ggtgattgta ggcaatcaca ccacagttaa tggcctggca
     4021 aataatgata agcgtgtgat aaacctcaca aaaatacgcc tagaagtggt gatatcgcta
     4081 gttttgtgtc ctttcgattt ttttcgtgtt gcttttttct gtggataaac ctttgaactt
     4141 tacattttgt tttcagaggc gcgcctctag aggatccccg ggattggcca aaggacccaa
     4201 aggtatgttt cgaatgatac taacataaca tagaacattt tcaggaggac ccttggctag
     4261 catgatgcgg ctgcgaggct cggggatgct gcgggacctg ctcctgcggt cgcccgcagg
     4321 agtatcggcg actctgcggc gggcacagcc cttggtcacc ctgtgccggc gtccccgagg
     4381 cgggggacgg ccggccgcgg gcccggctgc cgccgcgcga ctccacccgt ggtggggcgg
     4441 gggcggctgg ccggcggagc ccctcgcgcg gggcctgtcc agctctcctt cggagatctt
     4501 gcaggagctg ggcaagggga gcacgcatcc gcagcccggg gtgtcgccac ccgctgcccc
     4561 ggcggcgccc ggccccaagg acggccccgg ggagacggac gcgtttggca acagcgaggg
     4621 caaagagctg gtggcctcag gtgaaaataa aataaaacag ggtctgttag ctagcgcggc
     4681 agatgcgctg ttcgctacaa ttgctgaagc agcagcgaaa atacctgttc ataaatttat
     4741 tacagcactc aaatctacag gattgcgaac gtctgatccc aggttggcag cgtgtatggc
     4801 tatgttaaga ttaactcttg caacaacatc agatggtgtc atgctagaca aagatctttt
     4861 taaaaaatgt gttcagagca acattgcttt gttggcagca gcatttgcaa gagcgtttgt
     4921 gattcctgcc tttatggctt ttacctcagc cattgatgag ttatatgaaa gtgctaaaaa
     4981 gcagtctgga ggaaaggttg cagattatat tcctcaactg gccaaattca gtcccgattt
     5041 gtggggtgtg tctgtttgta cagtagatgg acagaggcat tctactggag ataccaaagt
     5101 tcccttctgt cttcagtcct gtgtaaaacc tttgaaatat gccattgctg ttaatgatct
     5161 tggaactgaa tatgtgcatc gatatgttgg aaaagagccg agtggactaa gattcaacaa
     5221 actatttttg aatgaagatg ataaaccaca taatcctatg gtaaatgctg gagcaattgt
     5281 tgtgacttca ctaataaagc aaggagtaaa taatgctgaa aaatttgact atgtcatgca
     5341 gtttttgaat aagatggctg gtaatgaata tgttggattc agtaatgcaa cgtttcagtc
     5401 tgaaagagaa agtggagatc gaaattttgc aataggatat tacttaaaag aaaagaagtg
     5461 ttttccagaa ggcacagaca tggttggtat attagacttc tacttccagc tgtgctccat
     5521 tgaagtgact tgtgaatcag ccagtgtgat ggctgcgaca ctggctaatg gtggtgcctg
     5581 cccaattact gctgcagcag cactgagcgc tgaagcagtt cgaaatacat tgagtttgat
     5641 gcattcctgt ggcatgtatg acttctcagg gcagtttgct ttccatgttg gtcttcctgc
     5701 aaaatctgga gttgctgggg gcattctttt agttgtcccc aatgttatgg gtatgatgtg
     5761 ctggtctcct cctctggata agatgggcaa cagtgttaag ggaattcact tttgtcacga
     5821 tcttgtttct ctgtgtaatt tccataacta tgataatttg gcagccgctg cagcaaaagc
     5881 tgatcctcga gcagcaggtg gtgatcaaag ggtaaagtca gtgataaatc ttttgtttgc
     5941 tgcatatact ggagatgtgt ctgcacttcg aagatttgct ttgtcagcta tggacatgga
     6001 acagcgggac tatgattcta gaacagcact ccatgtagct gctgcagagg gtcatgttga
     6061 agttgttaaa tttttgctgg aagcctgcaa agtaaaccct ttccccaagg acaggtggaa
     6121 taacactccc atggatgaag cactgcactt tggacaccat gatgtattta aaattctcca
     6181 agaataccaa gtccagtaca cacctcaagg agattctgac aacgggaagg aaaatcaaac
     6241 agtccataag aatcttgatg gattgttggg taccgctgtc tcatcctact ttcacctagt
     6301 taactgcttg tcttaaaatc tatgcttctc tttagtatct aaaattttcc tagaagctta
     6361 caagtatata aatggtctct tctcaataaa ggttgtatat ttattcatct tattgaatct
     6421 gccatttcct cgtttttgcg agtttatata ccttccaatt ttctttctat tgtattttca
     6481 acttctaatt ttaattcagg gaaactgctg taccggtaga aaaaatgagt aaaggagaag
     6541 aacttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca
     6601 aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat
     6661 ttatttgcac tactggaaaa ctacctgttc catgggtaag tttaaacata tatatactaa
     6721 ctaaccctga ttatttaaat tttcagccaa cacttgtcac tactttctgt tatggtgttc
     6781 aatgcttctc gagataccca gatcatatga aacggcatga ctttttcaag agtgccatgc
     6841 ccgaaggtta tgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac
     6901 gtaagtttaa acagttcggt actaactaac catacatatt taaattttca ggtgctgaag
     6961 tcaagtttga aggtgatacc cttgttaata gaatcgagtt aaaaggtatt gattttaaag
     7021 aagatggaaa cattcttgga cacaaattgg aatacaacta taactcacac aatgtataca
     7081 tcatggcaga caaacaaaag aatggaatca aagttgtaag tttaaacatg attttactaa
     7141 ctaactaatc tgatttaaat tttcagaact tcaaaattag acacaacatt gaagatggaa
     7201 gcgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt
     7261 taccagacaa ccattacctg tccacacaat ctgccctttc gaaagatccc aacgaaaaga
     7321 gagaccacat ggtccttctt gagtttgtaa cagctgctgg gattacacat ggcatggatg
     7381 aactatacaa atagcattcg tagaattcca actgagcgcc ggtcgctacc cagctttctt
     7441 gtacaaagtg gtgatatctg agctccgcat cggccgctgt catcagatcg ccatctcgcg
     7501 cccgtgcctc tgacttctaa gtccaattac tcttcaacat ccctacatgc tctttctccc
     7561 tgtgctccca ccccctattt ttgttattat caaaaaactt ctcttaattt ctttgttttt
     7621 tagcttcttt taagtcacct ctaacaatga aattgtgtag attcaaaaat agaattaatt
     7681 cgtaataaaa agtcgaaaaa aattgtgctc cctcccccca ttaataataa ttctatccca
     7741 aaatctacac aatgttctgt gtacacttct tatgtttttt acttctgata aatttttttg
     7801 aaacatcata gaaaaaaccg cacacaaaat accttatcat atgttacgtt tcagtttatg
     7861 accgcaattt ttatttcttc gcacgtctgg gcctctcatg acgtcaaatc atgctcatcg
     7921 tgaaaaagtt ttggagtatt tttggaattt ttcaatcaag tgaaagttta tgaaattaat
     7981 tttcctgctt ttgctttttg gggtttcccc tattgtttgt caagatttcg aggacggcgt
     8041 ttttcttgct aaaatcacaa gtattgatga gcacgatgca agaaagatcg gaagaaggtt
     8101 tgggtttgag gctcagtgga aggtgagtag aagttgataa tttgaaattg gagtagtgtc
     8161 tatggggttt ttgccttaaa tgacagaata cattcccaat ataccaaaca taactgtttc
     8221 tactagtcgg ccgtacgggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     8281 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     8341 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     8401 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     8461 cgtaaggaga aaataccgca tcaggcggcc ttaattaagg gcctcgtgat acgcctattt
     8521 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     8581 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     8641 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     8701 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     8761 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     8821 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     8881 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     8941 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     9001 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     9061 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     9121 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     9181 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     9241 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     9301 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     9361 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     9421 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     9481 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     9541 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     9601 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     9661 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     9721 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     9781 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     9841 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     9901 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     9961 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
    10021 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
    10081 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
    10141 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
    10201 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
    10261 cttgagcgtc gatttttgtg at
//
