LOCUS       sequence-315442-        7589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Untagged WDR59 cDNA expression vector for transfections.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7589)
  AUTHORS   Orozco JM, Krawczyk PA, Scaria SM, Cangelosi AL, Chan SH, Kunchok T,
            Lewis CA, Sabatini DM
  TITLE     Dihydroxyacetone phosphate signals glucose availability to mTORC1.
  JOURNAL   Nat Metab. 2020 Sep;2(9):893-901. doi: 10.1038/s42255-020-0250-5. 
            Epub 2020 Jul 27.
  PUBMED    32719541
REFERENCE   2  (bases 1 to 7589)
  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"; doi: 
            "10.1038/s42255-020-0250-5"; journalName: "Nat Metab"; date: 
            "2020-09"; volume: "2"; issue: "9"; pages: "893-901"
COMMENT     Sequence Label: WDR59 in pRK5
FEATURES             Location/Qualifiers
     source          1..7589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        435..453
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     435..452
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     466..485
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      532..541
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      536..545
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3496..3505
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3509..3643
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3546..3565)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3600..3619
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3712..4069
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3712..3903
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3735..3755)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3920..4055
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4089..4106)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4089..4105)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4098..4120)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4318..4773
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4405..4424)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4615..4636
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4790..4809)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4909..4931
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4969..4987)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5055..5159
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5160..5228,5229..6020)
                     /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(5378..5397)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6191..6779
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6680..6699
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6933..6950
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7067..7088
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7103..7108,7109..7126,7127..7133)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7141..7157
                     /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     7146..7168
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7165..7181
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7165..7181
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7210..7589
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      241 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg cggccgggaa
      301 cggtgcattg gaacgcggat tccccgtgcc aagagtgacg taagtaccgc ctatagagtc
      361 tataggccca cccccttggc ttcgttagaa cgcggctaca attaatacat aaccttatgt
      421 atcatacaca tacgatttag gtgacactat agaataacat ccactttgcc tttctctcca
      481 caggtgtcca ctcccaggtc caactgcacc tcggttctat cgattgaatt cgccaccatg
      541 gcggcgcgat ggagcagcga aaacgtggtt gtagagttcc gtgactccca ggcaactgcg
      601 atgtctgtgg actgtcttgg gcagcatgca gtgctttctg gccgcagatt cttatacatc
      661 gtcaatctag atgccccttt cgaaggtcac cgaaagatct ctcgccagag caaatgggac
      721 attggagctg tgcagtggaa tcctcatgac agctttgcac actattttgc ggcttcgagt
      781 aaccaacgag tagaccttta caagtggaaa gacggcagtg gggaagttgg cacaacctta
      841 caaggccaca ctcgtgtcat cagcgacttg gactgggcgg tgtttgagcc tgacctcctg
      901 gttaccagct ctgtggacac ctacatctac atttgggata tcaaagacac aaggaaacct
      961 actgttgcac tgtctgctgt tgcgggtgcc tcccaggtca aatggaataa aaaaaatgct
     1021 aactgccttg ccaccagcca tgacggcgat gtgcggatat gggataagag gaaacccagt
     1081 acagcagtgg aatatctagc cgcccacctc tccaaaatcc atggcctgga ctggcaccca
     1141 gacagcgagc acattcttgc tacctccagt caagacaatt ctgtgaagtt ctgggattac
     1201 cgccagcctc ggaaatacct caatattctt ccttgccagg tgcctgtctg gaaggccaga
     1261 tacacacctt tcagcaatgg attggtgact gtgatggttc cccagctgcg gagggaaaac
     1321 agccttctcc tgtggaatgt ctttgacttg aacaccccag tccacacctt cgtggggcat
     1381 gatgatgtgg tcctggagtt ccagtggagg aagcagaagg aagggtccaa ggactatcaa
     1441 ctggtgacgt ggtcccggga tcagaccttg agaatgtggc gggtggattc ccagatgcag
     1501 aggctttgtg caaatgacat attagatggt gttgatgagt tcattgagag tatttccctt
     1561 ctgccggaac ctgagaagac cctgcacact gaagatacag atcaccagca cactgcaagc
     1621 catggggagg aagaagccct aaaagaagat ccccctagaa atctcctgga agagaggaaa
     1681 tcagatcaac tggggctgcc tcagaccttg cagcaggagt tctccctgat caatgtgcaa
     1741 atccggaatg tcaatgtgga gatggatgcg gcagacagga gctgcacagt gtctgtgcac
     1801 tgcagcaacc atcgtgtcaa gatgctggtg aagttccctg cacagtaccc aaacaacgcc
     1861 gccccttcct tccagtttat taaccccaca accatcacat ccaccatgaa agctaagctg
     1921 ctgaagatcc tgaaggacac agccctgcag aaagtgaagc gtggccagag ctgcctggag
     1981 ccctgcctgc gccagctcgt ctcctgcctt gagtcctttg tgaaccagga agacagcgct
     2041 tccagcaacc cgtttgcact ccccaactct gtcactcccc ccttaccgac gtttgcgcgg
     2101 gtgaccacgg cttacgggtc gtaccaggac gccaacattc cctttcctag gacttctggg
     2161 gccaggttct gcggagcagg ttacctggta tatttcacaa ggcccatgac aatgcatcgg
     2221 gcggtgtctc ccacagagcc tactccgaga tctctctcag ccttgtctgc ttatcacact
     2281 ggcttgatcg cgcccatgaa gatccgcaca gaggcccctg ggaaccttcg tttatacagt
     2341 gggagcccca ctcgcagcga gaaagagcag gtctccatca gctccttcta ctacaaggag
     2401 cggaaatcaa gacgatggaa aagtaagcgt gagggatcag actctggcaa tcgacagatc
     2461 aaggctgctg ggaaagtcat catccaggat attgcttgcc tcctgcctgt tcacaaatcg
     2521 ctgggagagc tgtacatatt gaatgtgaat gatattcagg aaacatgtca gaagaatgcc
     2581 gcctctgcct tgctcgttgg aagaaaggat cttgtccagg tttggtcgct ggctacggta
     2641 gctacagatc tttgccttgg tccgaaatct gacccagatt tggaaacacc ctgggctcga
     2701 catccatttg ggcggcagct gctggagtcc ctgttggctc actattgccg gctccgggat
     2761 gttcagacac tggcgatgct ctgtagcgtg tttgaagccc agtctcggcc tcaggggcta
     2821 ccaaacccct ttgggccttt tcctaaccgt tcttctaatc ttgtggtgtc ccatagtcga
     2881 tatcctagct ttacctcttc tggttcctgc tccagtatgt cagacccagg gctcaacact
     2941 ggcggctgga acatagcggg aagagaggca gagcacttgt cctccccttg gggagaatcc
     3001 tcaccagaag agctccgctt tgggagtctg acctacagtg atccccgtga gcgagaacgc
     3061 gaccagcatg ataaaaataa aaggctcctg gaccccgcca atacccagca atttgatgac
     3121 tttaagaaat gctatgggga aatcctctac cgttggggtc tgagagagaa gcgagctgaa
     3181 gtgttgaagt ttgtctcctg tcctcctgac cctcacaaag ggatcgagtt cggcgtgtac
     3241 tgcagccact gccggagtga ggtccgtggc acgcagtgtg ccatctgcaa aggcttcacg
     3301 ttccagtgtg ccatctgtca cgtggctgtg cggggatcgt ccaatttctg cctgacctgt
     3361 gggcacggtg gccacaccag ccacatgatg gagtggtttc ggacccagga ggtgtgtccc
     3421 accgggtgtg ggtgccactg cctgcttgaa agcactttct gagcggccgc taagtaagta
     3481 aggatcccca gcttggccgc catggcccaa cttgtttatt gcagcttata atggttacaa
     3541 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3601 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcgggaatt aattcggcgc
     3661 agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct tctgaggcgg
     3721 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3781 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3841 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3901 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3961 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4021 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctgttaac
     4081 agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     4141 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     4201 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
     4261 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     4321 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4381 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4441 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4501 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     4561 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4621 tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag
     4681 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4741 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     4801 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     4861 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4921 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     4981 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     5041 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5101 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5161 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5221 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5281 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5341 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5401 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5461 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5521 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5581 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5641 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5701 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5761 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5821 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5881 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5941 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6001 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6061 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6121 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6181 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6241 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6301 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6361 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6421 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6481 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6541 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6601 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6661 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6721 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6781 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6841 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     6901 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     6961 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     7021 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     7081 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     7141 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aattaattcg
     7201 agctcgcccg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     7261 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     7321 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     7381 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     7441 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     7501 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     7561 tctacgtatt agtcatcgct attaccatg
//
