LOCUS       sequence-129576-        7678 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 7678)
  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 7678)
  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 tubulin myc
FEATURES             Location/Qualifiers
     source          1..7678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    55..76
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(91..96,97..114,115..121)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    129..145
                     /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             307..821
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(921..1520)
                     /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(1004..1024)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1501..1520
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(1541..2040)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1589..1608)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     1982..2005
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     CDS             complement(2057..2086)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3489..3905)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3505..3521)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3537..3559)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3972..4313
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             4377..4893
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(5093..5112)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5240..5262)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5456..5475)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5575..5597
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5635..5653)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5721..5825
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5826..5894,5895..6686)
                     /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(6044..6063)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6857..7445
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7346..7365
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7599..7616
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
       61 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      121 gtgtggaatt gtgagcggat aacaatttca caattctccc cagctctttc tgcgctgtcc
      181 ataatgaact atttcaggag tattggagta tgtggattca aaacagtggc taattgagtc
      241 aggattactt aatattcgac tcctaccctc agaatgttgt tctccaactt caagaaactg
      301 ttacccatga aagacccccg ctgacgggta gtcaatcact cagaggagac cctcccaagg
      361 aacagcgaga ccacaagtcg gatgcaactg caagagggtt tattggatac acgggtaccc
      421 gggcgacgca gtctatcgga ggactggcgc gccgagtgag gggttgtggg ctcttttatt
      481 gagctcgggg agcagaagcg cgcgaacaga agcgagaagc gaactgattg gttagttcaa
      541 ataaggcaca gggtcatttc aggtccttgg ggcaccctgg aaacatctga tggttctcta
      601 gaaactgctg agggcgggac cgcatctggg gaccatctgt tcttggccct gagccggggc
      661 aggaactgct taccacagat atcctgtttg gcccatattc tgctgtctct ctgttcctaa
      721 ccttgatctg aacttctcta ttctcagtta tgtattttcc atgccttgca aaatggcgtt
      781 acttaagcta gcttgccaaa cctacaggtg gggtctttca ttcccccctt tttctggaga
      841 ctaaataaaa tcttttattt tatcgatgca tggggtcgtg cgctcctttc ggtcgggcgc
      901 tgcgggtcgt ggggcgggcg tcaggcaccg ggcttgcggg tcatgcacca ggtgcgcggt
      961 ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta gaaggggagg
     1021 ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc cgcctccact
     1081 ccggggagca cgacggcgct gcccagaccc ttgccctggt ggtcgggcga gacgccgacg
     1141 gtggccagga accacgcggg ctccttgggc cggtgcggcg ccaggaggcc ttccatctgt
     1201 tgctgcgcgg ccagccggga accgctcaac tcggccatgc gcgggccgat ctcggcgaac
     1261 accgcccccg cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc gccgtcgtcc
     1321 gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc ttgcagctcg
     1381 gtgacccgct cgatgtggcg gtccggatcg acggtgtggc gcgtggcggg gtagtcggcg
     1441 aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt cgcgggtggc gaggcgcacc
     1501 gtgggcttgt actcggtcat ggtaagcttg ggctgcaggt cgaaaggccc ggagatgagg
     1561 aagaggagaa cagcgcggca gacgtgcgct tttgaagcgt gcagaatgcc gggcctccgg
     1621 aggaccttcg ggcgcccgcc ccgcccctga gcccgcccct gagcccgccc ccggacccac
     1681 cccttcccag cctctgagcc cagaaagcga aggagcaaag ctgctattgg ccgctgcccc
     1741 aaaggcctac ccgcttccat tgctcagcgg tgctgtccat ctgcacgaga ctagtgagac
     1801 gtgctacttc catttgtcac gtcctgcacg acgcgagctg cggggcgggg gggaacttcc
     1861 tgactagggg aggagtagaa ggtggcgcga aggggccacc aaagaacgga gccggttggc
     1921 gcctaccggt ggatgtggaa tgtgtgcgag gccagaggcc acttgtgtag cgccaagtgc
     1981 ccagcggggc tgctaaagcg catgctccag actgccttgg gaaaagcgcc tcccctaccc
     2041 ggtagaattc ttaattgaga tcttcctcag aaatgagctt ttgctccata ccaccaccac
     2101 cagcggccgc ctgctcctgg ggcacccagg agatgtagtc tggccgtgtg gccgcatggt
     2161 actcatcgat gagctgctgc acaatctccc tggatgtgtc catctcatca aagttgtcct
     2221 tgaacatgtc ctccttgcgg aactgctcca ggaaggcctc ccgcttacgc agcttgtcat
     2281 actggcgaca ggttctctcg aagagcgagg agatgctggt gtggttggcc atcatgagcc
     2341 cgctgacccg gtgggccgag ggcaggtagg gagacttcct cgacagggcc acctggatgc
     2401 tggcggggcc ccacgggatg aagttggcca acttccgttc ccggatcctc tgcaagctct
     2461 tgtggacctg ggtggggtcc acctctccct ggatgatgtt gaggatggcg atgtagcagt
     2521 ggttggtctg gcggtctcgg cctgtggaca ccatcacgtt cttgggctgc agcagccgcc
     2581 tcatgacatc caggaccgtg gtcttcctca cgctggccac tgactggtcc gtagtgagag
     2641 gggtgtagcc ggtcatgagg aagtggagcc gtggggtggg aatgagcgag gcgatgaggc
     2701 cgatgaggtc attgttcatg tagccagggt agcgcagggt ggtggtgctg gctgacatga
     2761 tggtagacac cagctggttg atctgggaga aggatgggtt ctggatgtgc aggcggtctg
     2821 tggcaatccg gttcagggct gtgttgtcca gcaccaccac acagtctgca ttctgcgtca
     2881 gcctcttgag tgtgaggagt gaattgtaag gctggaccac cacatcgctc atctcgtcct
     2941 ggttgggaaa cactgagtat gtctgcacca gcttcttagg atacctgtca ttcagccgtt
     3001 ctaagaggta ggaacccagt ccagagcctg tccccccagc aatggagtga cacagcacaa
     3061 agccctctag actgtcacta ccatctgcct cccggtctat gatgtcaaaa atgtcctcat
     3121 ggatcttttc tccctgggag aatccgctgg cccagttgtt gccagctcct cctccatgtt
     3181 ccgacaggta gatgttctct gggttgtaga gcttggcata gggggagttg aggatggagt
     3241 ggatcacccg gggttccaag tccagcagca cggcccgggg gatgtagtgc tcatcgtctg
     3301 cctggtagaa aaagacgtcc ttgcggtcag tgccctcggt ggcgaactcc tccacgatgc
     3361 cctcggggct gataccatgc tcggcgcaca gctgtttcca gaactcgaac ccaatctgat
     3421 tgccgcactg gcccaactgt agggtgatga tttccctcgg catggtgccg gtcgagagat
     3481 ctaattccgg cgcctagaga aggagtgagg gctggataaa gggaggatcg aggcggggtc
     3541 gaacgaggag gttcaagggg gagagacggg gcggatggag gaagaggagg cggaggctta
     3601 gggtgtacaa agggcttgac ccagggaggg gggtcaaaag ccaaggcttc ccaggtcacg
     3661 atgtagggga cctggtctgg gtgtccatgc gggccaggtg aaaagacctt gatcttaacc
     3721 tgggtgatga ggtctcggtt aaaggtgccg tctcgcggcc atccgacgtt aaaggttggc
     3781 cattctgcag agcagaaggt aacccaacgt ctcttcttga catctaccga ctggttgtga
     3841 gcgatccgct cgacatcttt ccagtgacct aaggtcaaac ttaagggagt ggtaacagtc
     3901 tggccctaat tttcagacaa atacagaaac acagtcagac agagacaaca cagaacgatg
     3961 ctgcagcgct gcagcagaca agacgcgcgg cttcggttcc aaaccgaaag caaaaattca
     4021 gacggaggcg ggaactgttt taggttctcg tctcctacca gaaccacata tcctgacggg
     4081 gtcggattcc acatcgactc ccttcctcag gtcgggccac aaaaacggcc cccaaagtcc
     4141 ctgggacgtc tcccagggtt gcggccgggt gttcagaact cgtcagttcc accacgggtc
     4201 cgccagatac agagctagtt agctaactag tacagacgca ggcgcaaaca ttagatgccg
     4261 gcacaaacac gcacaaagac agagacagac acgaaacgac cgctggccag cttacctccc
     4321 ggcggggggg tcggtggtcc ctgggcaggg gtctccaaat ctcggtggaa cctccaaatg
     4381 aaagaccccc gaggtgggca gtcaatcaat ctgaggagac cctcccaagg atcagcgagt
     4441 ccacgattcg gatgcaaaca gcaagaggct ttattgggaa tacgggtacc cgggcgacgc
     4501 agtctatcgg aggactggcg cgccgagtga ggggttgtgg gctcttttat tgagctcggg
     4561 gagcagaagc gcgcgaacag aagcgagaag cgaactgatt ggttagttca aataaggcac
     4621 agggtcattt caggtccttg gggcaccctg gaaacatctg atggttctct agaaactgct
     4681 gagggcggga ccgcatctgg ggaccatctg ttcttggccc tgagccgggg caggaactgc
     4741 ttaccacaga tatcctgttt ggcccatatt ctgctgtctc tctgttccta accttgatct
     4801 gaacttctct attctcagtt atgtattttc catgccttgc aaaatggcgt tacttaagct
     4861 agcttgccaa acctacaggt ggggtctttc attcccccct ttttctggag actaaataaa
     4921 atcttttatt ttatctatgg ctcgtactct ataggcttca gctggtgata ttgttgagtc
     4981 aaaactagag cctggaccac tgatatcctg tctttaacaa attggactaa tcgcctctac
     5041 gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc
     5101 gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc
     5161 ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat
     5221 gcaccattcc ttgcgccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     5281 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     5341 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     5401 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     5461 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     5521 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     5581 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     5641 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5701 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5761 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5821 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5881 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5941 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     6001 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6061 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6121 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6181 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6241 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6301 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6361 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6421 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6481 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6541 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6601 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6661 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6721 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6781 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6841 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6901 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6961 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     7021 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7081 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7141 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7201 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7261 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7321 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7381 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7441 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7501 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     7561 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     7621 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatg
//
