LOCUS       sequence-130707-        4979 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 4979)
  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 4979)
  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 DEPTOR (PDZ domain)
FEATURES             Location/Qualifiers
     source          1..4979
                     /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      1279..1288
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1292..1426
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1329..1348)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1383..1402
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1495..1852
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1495..1686
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1518..1538)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1703..1838
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1872..1889)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1872..1888)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1881..1903)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2101..2556
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2188..2207)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2398..2419
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2573..2592)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2692..2714
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2752..2770)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2838..2942
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2943..3011,3012..3803)
                     /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(3161..3180)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3974..4562
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4463..4482
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4716..4733
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4850..4871
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4886..4891,4892..4909,4910..4916)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4924..4940
                     /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     4929..4951
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4948..4964
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4948..4964
                     /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 gacggctccg tatgcaagga agacattcac gattgttggt
     1021 gacgcggttg gctggggttt tgtggtgcga ggaagtaagc catgccacat ccaggctgta
     1081 gaccccagtg gccctgcagc cgcagcagga atgaaggtct gtcagtttgt cgtctctgtc
     1141 aacgggctca atgtcctgca tgtagactac cggaccgtga acaatctgat tctgacgggc
     1201 ccacggacga ttgtcatgga agtcatggag gagttagagt gctgagcggc cgctaagtaa
     1261 gtaaggatcc ccagcttggc cgccatggcc caacttgttt attgcagctt ataatggtta
     1321 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     1381 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcggga attaattcgg
     1441 cgcagcacca tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg
     1501 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     1561 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     1621 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     1681 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     1741 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     1801 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctgtt
     1861 aacagcttgg cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     1921 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     1981 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg
     2041 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     2101 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2161 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2221 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2281 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     2341 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2401 gactcttgtt ccaaactgga acaacactca accctatctc gggctattct tttgatttat
     2461 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     2521 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     2581 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     2641 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2701 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     2761 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     2821 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2881 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2941 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3001 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3061 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3121 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3181 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3241 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3301 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3361 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3421 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3481 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3541 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     3601 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3661 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3721 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3781 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     3841 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3901 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3961 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4021 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4081 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4141 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4201 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4261 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4321 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4381 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4441 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4501 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4561 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4621 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     4681 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     4741 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     4801 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     4861 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     4921 gaattgtgag cggataacaa tttcacacag gaaacagcta tgacatgatt acgaattaa
//
