LOCUS       sequence-132131-        5948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5948)
  AUTHORS   Peterson TR, Laplante M, Thoreen CC, Sancak Y, Kang SA, Kuehl WM, 
            Gray NS, Sabatini DM
  TITLE     DEPTOR is an mTOR inhibitor frequently overexpressed in multiple 
            myeloma cells and required for their survival.
  JOURNAL   Cell. 2009 May 29;137(5):873-86. doi: 10.1016/j.cell.2009.03.046. 
            Epub 2009 May 14.
  PUBMED    19446321
REFERENCE   2  (bases 1 to 5948)
  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.2009.03.046"; journalName: "Cell"; date: 
            "2009-05-29- 29"; volume: "137"; issue: "5"; pages: "873-86"
COMMENT     Sequence Label: pRK5 FLAG human DEPTOR
FEATURES             Location/Qualifiers
     source          1..5948
                     /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"
     CDS             937..960
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      2248..2257
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2261..2395
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2298..2317)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2352..2371
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2464..2821
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2464..2655
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2487..2507)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2672..2807
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2841..2858)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2841..2857)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2850..2872)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3070..3525
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3157..3176)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3367..3388
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3542..3561)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3661..3683
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3721..3739)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3807..3911
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3912..3980,3981..4772)
                     /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(4130..4149)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4943..5531
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5432..5451
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5685..5702
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5819..5840
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5855..5860,5861..5878,5879..5885)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5893..5909
                     /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     5898..5920
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5917..5933
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5917..5933
                     /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 attcaccatg gaaatggact acaaggatga cgatgacaag
      961 ggtggtggtg gtggtgcgtc gacgatggag gagggcggca gcactggcag tgctggcagt
     1021 gacagcagca ccagcgggag tggcggggcg cagcaaaggg agctggagcg catggctgag
     1081 gtcttggtca ccggggaaca gctacggctc aggctgcatg aagaaaaggt tattaaagat
     1141 agacgtcatc atctcaagac ctacccaaac tgttttgtcg caaaagaact gattgactgg
     1201 ctgattgaac acaaagaggc ttctgacaga gagacggcaa ttaaactcat gcagaaatta
     1261 gcagaccggg gcattattca ccatgtgtgt gatgagcata aggaattcaa ggatgtcaaa
     1321 ctcttctacc gctttagaaa ggatgacggc accttcccat tggataatga agtgaaggcc
     1381 tttatgagag gacagaggct atatgaaaag ctgatgagcc ctgaaaacac actcctgcag
     1441 cccagggagg aggaaggggt caagtatgag cgcaccttca tggcatctga attcctggac
     1501 tggctggttc aggaaggtga ggccaccacg aggaaagagg cagagcagct ttgccaccgg
     1561 cttatggagc atggcatcat ccagcatgtg tccagcaagc acccatttgt ggacagcaat
     1621 cttctctacc agttcagaat gaacttccgg cggaggcgaa gactgatgga gctgctcaat
     1681 gaaaagtccc cctcctccca ggaaactcat gacagtccct tctgcctgag gaagcagagc
     1741 catgacaatc ggaaatctac cagctttatg tcagtgagcc ccagcaagga gatcaagatc
     1801 gtgtctgcag tgaggagaag cagcatgagc agctgtggca gcagcggcta cttcagcagc
     1861 agccccaccc tcagcagcag cccccctgtg ctctgcaacc ccaagtccgt gctgaagaga
     1921 cctgtcacct ctgaggaact ccttactccc ggggctccgt atgcaaggaa gacattcacg
     1981 attgttggtg acgcggttgg ctggggtttt gtggtgcgag gaagtaagcc atgccacatc
     2041 caggctgtag accccagtgg ccctgcagcc gcagcaggaa tgaaggtctg tcagtttgtc
     2101 gtctctgtca acgggctcaa tgtcctgcat gtagactacc ggaccgtgaa caatctgatt
     2161 ctgacgggcc cacggacgat tgtcatggaa gtcatggagg agttagagtg ctgagcggcc
     2221 gctaagtaag taaggatccc cagcttggcc gccatggccc aacttgttta ttgcagctta
     2281 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2341 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcgggaa
     2401 ttaattcggc gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac
     2461 cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2521 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     2581 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2641 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     2701 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     2761 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     2821 aaagctgtta acagcttggc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     2881 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     2941 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     3001 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     3061 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3121 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3181 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3241 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     3301 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3361 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt
     3421 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3481 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     3541 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     3601 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3661 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     3721 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     3781 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3841 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3901 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3961 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4021 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4081 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4141 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4201 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4261 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4321 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4381 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4441 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4501 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4561 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4621 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4681 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4741 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4801 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4861 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4921 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4981 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5041 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5101 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5161 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5221 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5281 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5341 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5401 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5461 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5521 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5581 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5641 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5701 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5761 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5821 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     5881 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gacatgatta
     5941 cgaattaa
//
