LOCUS       sequence-163585-        5245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  overexpression of CASTOR2 ACT2 domain (aa 171-end).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5245)
  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 5245)
  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 CASTOR2 ACT2
FEATURES             Location/Qualifiers
     source          1..5245
                     /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      1545..1554
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1558..1692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1595..1614)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1649..1668
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1761..2118
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1761..1952
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1784..1804)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1969..2104
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2138..2155)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2138..2154)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2147..2169)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2367..2822
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2454..2473)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2664..2685
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2839..2858)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2958..2980
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3018..3036)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3104..3208
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3209..3277,3278..4069)
                     /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(3427..3446)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4240..4828
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4729..4748
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4982..4999
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5116..5137
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5152..5157,5158..5175,5176..5182)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5190..5206
                     /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     5195..5217
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5214..5230
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5214..5230
                     /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 ggtccagagg ccagtcatcc acccactgtc cagcccgagc aacaggttct
     1081 gtgtcaccag cctggaccct gacacgctgc ctgctgttgc cacactcctc atggatgtca
     1141 tgttctactc caatggagtg aaggacccca tggccactgg ggatgactgc ggccacatcc
     1201 gcttcttctc cttctccctc atcgagggct acatctccct ggtgatggac gtgcagacgc
     1261 agcagaggtt tcctagtaac ttgctgttca caagcgcatc cggagagctc tggaagatgg
     1321 tccggattgg aggacagccc ctggggtttg atgagtgtgg catcgtggcc cagatctcag
     1381 agcccttggc tgctgcagac atcccagcct actacatcag tactttcaag tttgatcatg
     1441 cacttgtccc cgaagagaac atcaatggtg tcatcagtgc cctgaaggtc agccaagcag
     1501 agaagcactg agcggccgct aagtaagtaa ggatccccag cttggccgcc atggcccaac
     1561 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     1621 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     1681 catgtctgga tcgggaatta attcggcgca gcaccatggc ctgaaataac ctctgaaaga
     1741 ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     1801 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     1861 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     1921 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     1981 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     2041 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     2101 ggcctaggct tttgcaaaaa gctgttaaca gcttggcact ggccgtcgtt ttacaacgtc
     2161 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     2221 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     2281 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     2341 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     2401 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     2461 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2521 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     2581 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     2641 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     2701 tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     2761 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     2821 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     2881 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     2941 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     3001 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     3061 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3181 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3421 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3721 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     3781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4381 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4981 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     5041 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     5101 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     5161 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     5221 cagctatgac atgattacga attaa
//
