LOCUS       sequence-197113-        6199 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA tagged metap2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6199)
  AUTHORS   Gu X, Orozco JM, Saxton RA, Condon KJ, Liu GY, Krawczyk PA, Scaria 
            SM, Harper JW, Gygi SP, Sabatini DM
  TITLE     SAMTOR is an S-adenosylmethionine sensor for the mTORC1 pathway.
  JOURNAL   Science. 2017 Nov 10;358(6364):813-818. doi: 
            10.1126/science.aao3265.
  PUBMED    29123071
REFERENCE   2  (bases 1 to 6199)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2017-11-10- 10"; volume: "358"; 
            issue: "6364"; pages: "813-818"
COMMENT     Sequence Label: pRK5-HA-metap2
FEATURES             Location/Qualifiers
     source          1..6199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        144..523
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        524..727
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     678..698
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     724..748
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        958..976
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     958..975
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     989..1008
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1093..1102
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1105..1131
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      2629..2638
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2642..2776
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2679..2698)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2733..2752
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2845..3202
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2845..3036
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2868..2888)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3053..3188
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3222..3239)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3222..3238)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3231..3253)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3451..3906
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3538..3557)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3748..3769
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3923..3942)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4042..4064
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4102..4120)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4188..4292
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4293..4361,4362..5153)
                     /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(4511..4530)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5324..5912
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5813..5832
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6066..6083
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat
      121 tacgaattaa ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat
      181 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      241 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      301 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      361 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      421 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      481 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      541 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      601 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      661 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      721 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
      781 ccatagaaga caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg
      841 gattccccgt gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt
      901 ggcttcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
      961 taggtgacac tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag
     1021 gtccaactgc acctcggttc tatcgattga attcccgaac cgacagtcgg tctcttcacc
     1081 aaggccattc gcgccaccat gggctacccc tatgatgtgc ctgactacgc agatctcaat
     1141 ggtggtggtg gtgggtcgac gatggcgggt gtggaggagg tagcggcctc cgggagccac
     1201 ctgaatggcg acctggatcc agacgacagg gaagaaggag ctgcctctac ggctgaggaa
     1261 gcagccaaga aaaaaagacg aaagaagaag aagagcaaag ggccttctgc aggggaacag
     1321 gaacctgata aagaatcagg agcctcagtg gatgaagtag caagacagtt ggaaagatca
     1381 gcattggaag ataaagaaag agatgaagat gatgaagatg gagatggcga tggagatgga
     1441 gcaactggaa agaagaagaa aaagaagaag aagaagagag gaccaaaagt tcaaacagac
     1501 cctccctcag ttccaatatg tgacctgtat cctaatggtg tatttcccaa aggacaagaa
     1561 tgcgaatacc cacccacaca agatgggcga acagctgctt ggagaactac aagtgaagaa
     1621 aagaaagcat tagatcaggc aagtgaagag atttggaatg attttcgaga agctgcagaa
     1681 gcacatcgac aagttagaaa atacgtaatg agctggatca agcctgggat gacaatgata
     1741 gaaatctgtg aaaagttgga agactgttca cgcaagttaa taaaagagaa tggattaaat
     1801 gcaggcctgg catttcctac tggatgttct ctcaataatt gtgctgccca ttatactccc
     1861 aatgccggtg acacaacagt attacagtat gatgacatct gtaaaataga ctttggaaca
     1921 catataagtg gtaggattat tgactgtgct tttactgtca cttttaatcc caaatatgat
     1981 acgttattaa aagctgtaaa agatgctact aacactggaa taaagtgtgc tggaattgat
     2041 gttcgtctgt gtgatgttgg tgaggccatc caagaagtta tggagtccta tgaagttgaa
     2101 atagatggga agacatatca agtgaaacca atccgtaatc taaatggaca ttcaattggg
     2161 caatatagaa tacatgctgg aaaaacagtg ccgattgtga aaggagggga ggcaacaaga
     2221 atggaggaag gagaagtata tgcaattgaa acctttggta gtacaggaaa aggtgttgtt
     2281 catgatgata tggaatgttc acattacatg aaaaattttg atgttggaca tgtgccaata
     2341 aggcttccaa gaacaaaaca cttgttaaat gtcatcaatg aaaactttgg aacccttgcc
     2401 ttctgccgca gatggctgga tcgcttggga gaaagtaaat acttgatggc tctgaagaat
     2461 ctgtgtgact tgggcattgt agatccatat ccaccattat gtgacattaa aggatcatat
     2521 acagcgcaat ttgaacatac catcctgttg cgtccaacat gtaaagaagt tgtcagcagg
     2581 ggagatgact attaagcggc cgctaagtaa gtaaggatcc ccagcttggc cgccatggcc
     2641 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2701 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2761 ttatcatgtc tggatcggga attaattcgg cgcagcacca tggcctgaaa taacctctga
     2821 aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca
     2881 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2941 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3001 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3061 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3121 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3181 tggaggccta ggcttttgca aaaagctgtt aacagcttgg cactggccgt cgttttacaa
     3241 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     3301 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     3361 agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3421 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     3481 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3541 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3601 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3661 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3721 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3781 accctatctc gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3841 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     3901 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     3961 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4021 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4081 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     4141 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4201 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4261 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4321 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4381 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4441 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4501 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4561 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4621 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4681 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4741 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4801 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4861 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4921 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4981 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5041 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5101 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5161 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5221 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5281 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5341 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5401 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5461 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5521 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5581 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5641 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5701 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5761 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5821 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5881 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5941 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6001 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     6061 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     6121 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     6181 gcagtgagcg caacgcaat
//
