LOCUS       sequence-177201-        6190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  myc-TAG overexpression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6190)
  AUTHORS   Zoncu R, Bar-Peled L, Efeyan A, Wang S, Sancak Y, Sabatini DM
  TITLE     mTORC1 senses lysosomal amino acids through an inside-out mechanism 
            that requires the vacuolar H-ATPase.
  JOURNAL   Science. 2011 Nov 4;334(6056):678-83.
  PUBMED    22053050
REFERENCE   2  (bases 1 to 6190)
  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: "Science."; 
            date: "2011-11-4"; volume: "334(6056)"; pages: "678-83"
COMMENT     Sequence Label: pRK5-myc-PAT1
FEATURES             Location/Qualifiers
     source          1..6190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      950..959
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             959..988
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      2477..2486
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2490..2624
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2527..2546)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2581..2600
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2693..3050
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2693..2884
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2716..2736)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2901..3036
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3070..3087)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3070..3086)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3079..3101)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3299..3754
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3386..3405)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3596..3617
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3771..3790)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3890..3912
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3950..3968)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4036..4140
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4141..4209,4210..5001)
                     /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(4359..4378)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5172..5760
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5661..5680
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5914..5931
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6048..6069
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6084..6089,6090..6107,6108..6114)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6122..6138
                     /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     6127..6149
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6146..6162
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6146..6162
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
      901 cgattgaatt cccgaaccga cagtcggtct cttcaccaag gccattcgcg ccaccatgga
      961 gcaaaagctc atttctgagg aagatctcaa tggtggtggt ggtgggtcga cgatgtccac
     1021 gcagagactt cggaatgaag actaccacga ctacagctcc acggacgtga gccctgagga
     1081 gagcccgtcg gaaggcctca acaacctctc ctccccgggc tcctaccagc gctttggtca
     1141 aagcaatagc acaacatggt tccagacctt gatccacctg ttaaaaggca acattggcac
     1201 aggactcctg ggactccctc tggcggtgaa aaatgcaggc atcgtgatgg gtcccatcag
     1261 cctgctgatc ataggcatcg tggccgtgca ctgcatgggt atcctggtga aatgtgctca
     1321 ccacttctgc cgcaggctga ataaatcctt tgtggattat ggtgatactg tgatgtatgg
     1381 actagaatcc agcccctgct cctggctccg gaaccacgca cactggggaa gacgtgttgt
     1441 ggacttcttc ctgattgtca cccagctggg attctgctgt gtctattttg tgtttctggc
     1501 tgacaacttt aaacaggtga tagaagcggc caatgggacc accaataact gccacaacaa
     1561 tgagacggtg attctgacgc ctaccatgga ctcgcgactc tacatgctct ccttcctgcc
     1621 cttcctggtg ctgctggttt tcatcaggaa cctccgagcc ctgtccatct tctccctgtt
     1681 ggccaacatc accatgctgg tcagcttggt catgatctac cagttcattg ttcagaggat
     1741 cccagacccc agccacctcc ccttggtggc cccttggaag acctaccctc tcttctttgg
     1801 cacagcgatt ttttcatttg aaggcattgg aatggttctg cccctggaaa acaaaatgaa
     1861 ggatcctcgg aagttcccac tcatcctgta cctgggcatg gtcatcgtca ccatcctcta
     1921 catcagcctg gggtgtctgg ggtacctgca atttggagct aatatccaag gcagcataac
     1981 cctcaacctg cccaactgct ggttgtacca gtcagttaag ctgctgtact ccatcgggat
     2041 ctttttcacc tacgcactcc agttctacgt cccggctgag atcatcatcc ccttctttgt
     2101 gtcccgagcg cccgagcact gtgagttagt ggtggacctg tttgtgcgca cagtgctggt
     2161 ctgcctgaca tgcatcttgg ccatcctcat cccccgcctg gacctggtca tctccctggt
     2221 gggctccgtg agcagcagcg ccctggccct catcatccca ccgctcctgg aggtcaccac
     2281 cttctactca gagggcatga gccccctcac catctttaag gacgccctga tcagcatcct
     2341 gggcttcgtg ggctttgtgg tggggaccta tgaggctctc tatgagctga tccagccaag
     2401 caatgctccc atcttcatca attccacctg tgccttcata taggcggccg ctaagtaagt
     2461 aaggatcccc agcttggccg ccatggccca acttgtttat tgcagcttat aatggttaca
     2521 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2581 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcgggaat taattcggcg
     2641 cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttaggtacc ttctgaggcg
     2701 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2761 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2821 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2881 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2941 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3001 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctgttaa
     3061 cagcttggca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     3121 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     3181 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta
     3241 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     3301 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3361 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3421 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3481 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     3541 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3601 ctcttgttcc aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa
     3661 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3721 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     3781 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     3841 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3901 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     3961 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     4021 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4081 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4141 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4201 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4261 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4321 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4381 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4441 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4501 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     4561 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     4621 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     4681 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     4741 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     4801 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4861 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4921 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4981 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     5041 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5101 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5161 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5221 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5281 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5341 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5401 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5461 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5521 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5581 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5641 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5701 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5761 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5821 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     5881 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     5941 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     6001 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     6061 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     6121 attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac gaattaattc
     6181 gagctcgccc
//
