LOCUS       sequence-219919-        6823 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 6823)
  AUTHORS   Frias MA, Thoreen CC, Jaffe JD, Schroder W, Sculley T, Carr SA, 
            Sabatini DM
  TITLE     mSin1 is necessary for Akt/PKB phosphorylation, and its isoforms 
            define three distinct mTORC2s.
  JOURNAL   Curr Biol. 2006 Sep 19;16(18):1865-70. doi: 
            10.1016/j.cub.2006.08.001. Epub 2006 Aug 17.
  PUBMED    16919458
REFERENCE   2  (bases 1 to 6823)
  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.cub.2006.08.001"; journalName: "Curr Biol"; date: 
            "2006-09-19- 19"; volume: "16"; issue: "18"; pages: "1865-70"
COMMENT     Sequence Label: pMSCV-TAP
FEATURES             Location/Qualifiers
     source          1..6823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      8..596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     497..516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     750..767
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    884..905
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(920..925,926..943,944..950)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    958..974
                     /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)."
     LTR             1136..1650
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(1750..2349)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1833..1853)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2330..2349
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2370..2869)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2418..2437)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     2811..2834
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     CDS             complement(2926..3003)
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             complement(3010..3030)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(3067..3240)
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             complement(3241..3414)
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             complement(3463..3879)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3479..3495)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3511..3533)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3946..4287
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             4351..4867
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(5067..5086)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5214..5236)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5430..5449)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5549..5571
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5609..5627)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5695..5799
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5800..5868,5869..6660)
                     /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(6018..6037)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
       61 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
      121 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
      181 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      241 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      301 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
      361 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
      421 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
      481 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
      541 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
      601 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
      661 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
      721 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
      781 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
      841 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
      901 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
      961 tgagcggata acaatttcac aattctcccc agctctttct gcgctgtcca taatgaacta
     1021 tttcaggagt attggagtat gtggattcaa aacagtggct aattgagtca ggattactta
     1081 atattcgact cctaccctca gaatgttgtt ctccaacttc aagaaactgt tacccatgaa
     1141 agacccccgc tgacgggtag tcaatcactc agaggagacc ctcccaagga acagcgagac
     1201 cacaagtcgg atgcaactgc aagagggttt attggataca cgggtacccg ggcgacgcag
     1261 tctatcggag gactggcgcg ccgagtgagg ggttgtgggc tcttttattg agctcgggga
     1321 gcagaagcgc gcgaacagaa gcgagaagcg aactgattgg ttagttcaaa taaggcacag
     1381 ggtcatttca ggtccttggg gcaccctgga aacatctgat ggttctctag aaactgctga
     1441 gggcgggacc gcatctgggg accatctgtt cttggccctg agccggggca ggaactgctt
     1501 accacagata tcctgtttgg cccatattct gctgtctctc tgttcctaac cttgatctga
     1561 acttctctat tctcagttat gtattttcca tgccttgcaa aatggcgtta cttaagctag
     1621 cttgccaaac ctacaggtgg ggtctttcat tccccccttt ttctggagac taaataaaat
     1681 cttttatttt atcgatgcat ggggtcgtgc gctcctttcg gtcgggcgct gcgggtcgtg
     1741 gggcgggcgt caggcaccgg gcttgcgggt catgcaccag gtgcgcggtc cttcgggcac
     1801 ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag aaggggaggt tgcggggcgc
     1861 ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcc gcctccactc cggggagcac
     1921 gacggcgctg cccagaccct tgccctggtg gtcgggcgag acgccgacgg tggccaggaa
     1981 ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct tccatctgtt gctgcgcggc
     2041 cagccgggaa ccgctcaact cggccatgcg cgggccgatc tcggcgaaca ccgcccccgc
     2101 ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg ccgtcgtccg cgacccacac
     2161 cttgccgatg tcgagcccga cgcgcgtgag gaagagttct tgcagctcgg tgacccgctc
     2221 gatgtggcgg tccggatcga cggtgtggcg cgtggcgggg tagtcggcga acgcggcggc
     2281 gagggtgcgt acggccctgg ggacgtcgtc gcgggtggcg aggcgcaccg tgggcttgta
     2341 ctcggtcatg gtaagcttgg gctgcaggtc gaaaggcccg gagatgagga agaggagaac
     2401 agcgcggcag acgtgcgctt ttgaagcgtg cagaatgccg ggcctccgga ggaccttcgg
     2461 gcgcccgccc cgcccctgag cccgcccctg agcccgcccc cggacccacc ccttcccagc
     2521 ctctgagccc agaaagcgaa ggagcaaagc tgctattggc cgctgcccca aaggcctacc
     2581 cgcttccatt gctcagcggt gctgtccatc tgcacgagac tagtgagacg tgctacttcc
     2641 atttgtcacg tcctgcacga cgcgagctgc ggggcggggg ggaacttcct gactagggga
     2701 ggagtagaag gtggcgcgaa ggggccacca aagaacggag ccggttggcg cctaccggtg
     2761 gatgtggaat gtgtgcgagg ccagaggcca cttgtgtagc gccaagtgcc cagcggggct
     2821 gctaaagcgc atgctccaga ctgccttggg aaaagcgcct cccctacccg gtagaattcc
     2881 cggcggccgc gggatccatt cgctcgaggg accaccacca ccaccaagtg ccccggagga
     2941 tgagattttc ttaaagcggt tggctgctga gacggctatg aaattctttt tccatcttct
     3001 ctttagttca ccttgaaaat ataaattttc agaagcagtt gtaggtatat cacaattcgc
     3061 gtctactttc ggcgcctgag catcatttag ctttttagct tctgctaaaa ggttagcgct
     3121 ttggcttggg tcatctttta aactttggat gaaggcgttt cgttgttctt cgtttaagtt
     3181 aggtaaatgt aagatctcat agaacgcgtt ttgttgttct ttgttgaatt tgttgtctac
     3241 tttcggcgcc tgagcatcat ttagcttttt agcttctgct aaaaggttag cgctttggct
     3301 tgggtcatct tttaaacttt ggatgaaggc gtttcgttgt tcttcgttta agttaggtaa
     3361 atgtaagatc tcatagaacg cgttttgttg ttctttgttg aatttgttgt ccacggcttc
     3421 atcgtgttgc gcaaggcctg ccatggtgcc tgatctaatt ccggcgccta gagaaggagt
     3481 gagggctgga taaagggagg atcgaggcgg ggtcgaacga ggaggttcaa gggggagaga
     3541 cggggcggat ggaggaagag gaggcggagg cttagggtgt acaaagggct tgacccaggg
     3601 aggggggtca aaagccaagg cttcccaggt cacgatgtag gggacctggt ctgggtgtcc
     3661 atgcgggcca ggtgaaaaga ccttgatctt aacctgggtg atgaggtctc ggttaaaggt
     3721 gccgtctcgc ggccatccga cgttaaaggt tggccattct gcagagcaga aggtaaccca
     3781 acgtctcttc ttgacatcta ccgactggtt gtgagcgatc cgctcgacat ctttccagtg
     3841 acctaaggtc aaacttaagg gagtggtaac agtctggccc taattttcag acaaatacag
     3901 aaacacagtc agacagagac aacacagaac gatgctgcag cgctgcagca gacaagacgc
     3961 gcggcttcgg ttccaaaccg aaagcaaaaa ttcagacgga ggcgggaact gttttaggtt
     4021 ctcgtctcct accagaacca catatcctga cggggtcgga ttccacatcg actcccttcc
     4081 tcaggtcggg ccacaaaaac ggcccccaaa gtccctggga cgtctcccag ggttgcggcc
     4141 gggtgttcag aactcgtcag ttccaccacg ggtccgccag atacagagct agttagctaa
     4201 ctagtacaga cgcaggcgca aacattagat gccggcacaa acacgcacaa agacagagac
     4261 agacacgaaa cgaccgctgg ccagcttacc tcccggcggg ggggtcggtg gtccctgggc
     4321 aggggtctcc aaatctcggt ggaacctcca aatgaaagac ccccgaggtg ggcagtcaat
     4381 caatctgagg agaccctccc aaggatcagc gagtccacga ttcggatgca aacagcaaga
     4441 ggctttattg ggaatacggg tacccgggcg acgcagtcta tcggaggact ggcgcgccga
     4501 gtgaggggtt gtgggctctt ttattgagct cggggagcag aagcgcgcga acagaagcga
     4561 gaagcgaact gattggttag ttcaaataag gcacagggtc atttcaggtc cttggggcac
     4621 cctggaaaca tctgatggtt ctctagaaac tgctgagggc gggaccgcat ctggggacca
     4681 tctgttcttg gccctgagcc ggggcaggaa ctgcttacca cagatatcct gtttggccca
     4741 tattctgctg tctctctgtt cctaaccttg atctgaactt ctctattctc agttatgtat
     4801 tttccatgcc ttgcaaaatg gcgttactta agctagcttg ccaaacctac aggtggggtc
     4861 tttcattccc ccctttttct ggagactaaa taaaatcttt tattttatct atggctcgta
     4921 ctctataggc ttcagctggt gatattgttg agtcaaaact agagcctgga ccactgatat
     4981 cctgtcttta acaaattgga ctaatcgcct ctacgccgga cgcatcgtgg ccggcatcac
     5041 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
     5101 ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt
     5161 ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgc cgtcgtttta
     5221 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     5281 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     5341 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     5401 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     5461 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     5521 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     5581 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     5641 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5701 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5761 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5821 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5881 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5941 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     6001 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     6061 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6121 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6181 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6241 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6301 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6361 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6421 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6481 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6541 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6601 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6661 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6721 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6781 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aag
//
