LOCUS       sequence-163556-        5251 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  overexpression of CASTOR1 ACT2 domain (aa 169-end).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5251)
  AUTHORS   Chantranupong L, Scaria SM, Saxton RA, Gygi MP, Shen K, Wyant GA, 
            Wang T, Harper JW, Gygi SP, Sabatini DM
  TITLE     The CASTOR Proteins Are Arginine Sensors for the mTORC1 Pathway.
  JOURNAL   Cell. 2016 Mar 24;165(1):153-64. doi: 10.1016/j.cell.2016.02.035. 
            Epub 2016 Mar 10.
  PUBMED    26972053
REFERENCE   2  (bases 1 to 5251)
  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: 
            "10.1016/j.cell.2016.02.035"; journalName: "Cell"; date: 
            "2016-03-24- 24"; volume: "165"; issue: "1"; pages: "153-64"
COMMENT     Sequence Label: pRK5-HA CASTOR1 ACT2
FEATURES             Location/Qualifiers
     source          1..5251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      963..972
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             975..1001
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      1551..1560
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1564..1698
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1601..1620)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1655..1674
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1767..2124
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1767..1958
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1790..1810)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1975..2110
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2144..2161)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2144..2160)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2153..2175)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2373..2828
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2460..2479)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2670..2691
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2845..2864)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2964..2986
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3024..3042)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3110..3214
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3215..3283,3284..4075)
                     /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(3433..3452)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4246..4834
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4735..4754
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4988..5005
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5122..5143
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5158..5163,5164..5181,5182..5188)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5196..5212
                     /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     5201..5223
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5220..5236
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5220..5236
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgattga attcccgaac cgacagtcgg tctcttcacc aaggccattc
      961 gcgccaccat gggctacccc tatgatgtgc ctgactacgc agatctcaat ggtggtggtg
     1021 gtgggtcgac ggggcccagc cccacggtgc atcccatcca gagcccacag aaccgcttct
     1081 gtgtcctcac actggaccct gagacgcttc cagccatcgc caccaccctc atagatgtcc
     1141 tcttctactc gcacagcacc cccaaggagg cagcctctag cagtcctgaa cccagctcca
     1201 tcacgttctt tgccttctct ctcatcgagg gttatatctc cattgtcatg gatgctgaaa
     1261 cacagaaaaa gttccccagt gacctcctgc tgaccagctc ctcgggggag ctgtggagga
     1321 tggtgcgcat cggtggacag cccctgggct ttgatgaatg tggcatcgtg gcacagattg
     1381 caggtcccct ggctgccgct gacatctctg cctactacat cagcaccttc aacttcgacc
     1441 acgccctggt gcccgaggac ggtatcggca gcgtcatcga ggtcctccag cggcggcagg
     1501 aaggcctggc ttcctgagcg gccgctaagt aagtaaggat ccccagcttg gccgccatgg
     1561 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     1621 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     1681 tcttatcatg tctggatcgg gaattaattc ggcgcagcac catggcctga aataacctct
     1741 gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg gaatgtgtgt
     1801 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     1861 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     1921 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     1981 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     2041 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     2101 tttggaggcc taggcttttg caaaaagctg ttaacagctt ggcactggcc gtcgttttac
     2161 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     2221 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     2281 gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     2341 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     2401 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2461 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2521 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2581 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2641 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2701 caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2761 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     2821 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     2881 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     2941 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3001 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     3061 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3121 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3181 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3241 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3301 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3361 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3421 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3481 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3541 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3601 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3661 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3721 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     3781 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3841 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3901 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3961 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4021 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4081 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4141 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4201 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4261 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4321 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4381 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4441 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4501 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4561 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4621 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4681 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4741 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4801 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4861 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4921 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4981 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     5041 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     5101 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     5161 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     5221 aggaaacagc tatgacatga ttacgaatta a
//
