LOCUS       sequence-174792-        8226 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of WDR24.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8226)
  AUTHORS   Wolfson RL, Chantranupong L, Wyant GA, Gu X, Orozco JM, Shen K, 
            Condon KJ, Petri S, Kedir J, Scaria SM, Abu-Remaileh M, Frankel WN, 
            Sabatini DM
  TITLE     KICSTOR recruits GATOR1 to the lysosome and is necessary for 
            nutrients to regulate mTORC1.
  JOURNAL   Nature. 2017 Feb 15. doi: 10.1038/nature21423.
  PUBMED    28199306
REFERENCE   2  (bases 1 to 8226)
  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: "Nature. 
            2017 Feb 15. doi: 10.1038/nature21423."
COMMENT     Sequence Label: plc242-GFP-WDR24
FEATURES             Location/Qualifiers
     source          1..8226
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..470
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    535..734
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             935..1351
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1281..1303
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1319..1335
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1386..1395
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1392..1394,1395..1397,1398..2108)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1392..1394,1395..1397,1398..2108)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1437..1458)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1698..1717)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2045..2066
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2154..2174
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2187..2231
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(4804..4824)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4809..5138
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4989..5124
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5146..5544
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             5713..6182
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6312..6329)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6483..7071)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6563..6582)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7231..8022,8023..8091))
                     /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     7854..7873
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8092..8196)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
       61 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      121 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
      181 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
      241 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
      301 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
      361 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
      421 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
      481 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggcaagctg
      541 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
      601 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
      661 gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg
      721 tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct
      781 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
      841 gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg
      901 tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag
      961 tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt
     1021 caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg
     1081 gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc
     1141 acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc
     1201 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc
     1261 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
     1321 ttatccagcc ctcactcctt ctctaggcgc cggccggatc ccagtgtggt ggtacgctac
     1381 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1441 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1501 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     1561 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     1621 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1681 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1741 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1801 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     1861 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     1921 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1981 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     2041 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2101 tgtacaagtc cggaggggat cggtggagct ccaccggtgg cggccgctct agagagaatc
     2161 tttattttca gggcgccgcc aaattcaaag aaaccgctgc tgctaaattc gaacgccagc
     2221 acatggacag cggaggtgga ggtactagtc tcgagatgga gaagatgtcc cgtgtgacca
     2281 cagccctggg tggcagcgtg ctgacaggcc gcaccatgca ctgccacctg gatgctcccg
     2341 ccaatgccat cagtgtgtgc cgcgacgcag cccaggtggt cgtggcaggc cgtagcatct
     2401 tcaagatcta tgccatcgag gaggaacagt tcgtggaaaa gctgaacctg cgtgtggggc
     2461 gcaagccttc gcttaacctg agctgtgctg acgtggtctg gcaccagatg gatgagaacc
     2521 tgctggccac agcagccacc aatggcgtgg tggtcacgtg gaacctgggc cggccatccc
     2581 gcaacaagca ggaccagctg ttcacagaac acaagcgcac ggtaaacaaa gtctgcttcc
     2641 accccaccga agcccacgtg ctgctcagtg gctcccagga tggcttcatg aagtgctttg
     2701 acctccgcag aaaggactct gtcagcacct tctcgggcca gtcggagagc gtgcgggacg
     2761 tgcagttcag tatccgggac tacttcacct tcgcctccac ctttgagaac ggcaatgtgc
     2821 agctctggga catccggcgt cccgaccggt gcgagaggat gttcacagcc cacaacggac
     2881 ccgtcttctg ctgcgactgg caccccgagg acaggggctg gttggccact ggagggcgcg
     2941 acaagatggt gaaggtctgg gacatgacca cgcaccgtgc caaggagatg cactgtgtgc
     3001 agaccatcgc ctcggtggcc cgtgtgaagt ggcggccaga gtgccgccac cacctggcca
     3061 cgtgctccat gatggtggac cacaacatct atgtttggga cgtgcgccgg cccttcgtgc
     3121 cagctgccat gtttgaggaa caccgagacg tcaccacggg aattgcctgg cgccaccccc
     3181 acgacccctc cttcctgctg tctggctcca aggacagctc gctgtgccag cacctgttcc
     3241 gcgacgccag ccagcccgtc gagcgcgcca accctgaggg cctctgctac ggcctcttcg
     3301 gggacctggc cttcgccgcc aaggagagcc tcgtggctgc cgagtcgggg cgcaagccct
     3361 acactggcga ccggcgccac cccatcttct ttaagcgcaa gctggacccc gccgagccct
     3421 tcgcaggcct cgcctccagt gccctcagtg tctttgagac ggagccaggt ggcggcggca
     3481 tgcgctggtt tgtggacaca gctgagcgtt atgcgctggc tggccggcca ctggccgagc
     3541 tctgtgacca caacgcaaag gtggcccgag agcttggccg caaccaggtg gcgcaaacgt
     3601 ggaccatgct gcggatcatc tactgcagcc ctggcctagt gcccactgca aacctcaacc
     3661 acagtgtggg caagggtggc tcctgtggcc tcccgctcat gaacagtttc aacctgaagg
     3721 atatggcccc agggttgggc agtgagacgc ggctggaccg cagcaaagga gatgcgcgga
     3781 gcgacacagt tctgctcgac tcctcggcca cactcatcac caatgaggat aacgaggaaa
     3841 ccgagggcag cgacgtacct gccgactacc tgctgggtga cgtggaaggt gaggaggacg
     3901 agctgtacct gctggatccg gaacacgcgc accccgagga ccctgagtgc gtgctgccgc
     3961 aggaggcctt tccgctgcgc cacgagatcg tggacacgcc tcccgggccc gagcacctgc
     4021 aggacaaggc cgactccccg cacgtgagcg gcagcgaggc ggatgtggcc tccctggccc
     4081 ccgtggactc ctccttctcg ctcctgtctg tctcacacgc gctctacgac agccgcctgc
     4141 cgcccgactt cttcggcgtg ctggtgcgcg acatgctgca cttctacgct gagcagggcg
     4201 acgtgcagat ggctgtgtct gtgctcatcg tcctgggtga acgggtgcgc aaggacatcg
     4261 acgagcagac ccaggagcac tggtacactt cctacatcga cctgctgcag cgcttccgcc
     4321 tctggaacgt gtccaacgag gtggtcaagc tgagcaccag ccgcgccgtc agctgcctca
     4381 accaggcctc caccaccctg cacgtcaact gcagccactg caagcggccc atgagcagcc
     4441 ggggctgggt ctgcgacagg tgccaccgct gcgccagcat gtgtgcggtc tgccaccacg
     4501 tagtcaaggg tctcttcgtg tggtgccagg gctgcagcca cggcggccac ctgcagcaca
     4561 tcatgaagtg gctggaaggc agctcccact gtcccgcagg ctgcggccac ctctgcgagt
     4621 actcctgaga attctgcagt cgacggtacc gcgggcccgg gatccaccgg atctagataa
     4681 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     4741 cacctccccc tgaacctgaa acataaaatg aatgcaattc gccagcacag tggtcgaccc
     4801 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4861 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4921 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4981 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5041 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     5101 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttcatggc caagcctttg
     5161 tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc
     5221 tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt
     5281 gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct
     5341 gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc
     5401 atcttgagcc cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa
     5461 gccatagtga aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc
     5521 tctggttatg tgtgggaggg ctaagatcga taaaataaaa gattttattt agtctccaga
     5581 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat
     5641 tttgcaaggc atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa
     5701 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     5761 gctcagggcc aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     5821 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc
     5881 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     5941 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     6001 cccgagctca ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact
     6061 gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt
     6121 ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt
     6181 cacatgcagc atgtatcaaa attaatttgg ttttttttct taagtattta cattaaatgg
     6241 ccatagttgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6301 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6361 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6421 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6481 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6541 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6601 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6661 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6721 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6781 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6841 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6901 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6961 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7021 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     7081 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     7141 gtcatgagat tatcaaaaag gatcttcacc tagatccttt tgcggccgca aatcaatcta
     7201 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7261 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7321 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7381 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7441 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7501 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7561 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7621 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7681 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7741 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7801 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7861 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7921 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7981 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8041 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8101 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8161 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8221 tgcagc
//
