LOCUS       Exported                8601 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  expresses mir-17-92.
ACCESSION   .
VERSION     .
KEYWORDS    MSCV-mir-17-92
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8601)
  AUTHORS   Olive V, Sabio E, Bennett MJ, De Jong CS, Biton A, McGann JC, 
            Greaney SK, Sodir NM, Zhou AY, Balakrishnan A, Foth M, Luftig MA, 
            Goga A, Speed TP, Xuan Z, Evan GI, Wan Y, Minella AC, He L
  TITLE     A component of the mir-17-92 polycistronic oncomir promotes 
            oncogene-dependent apoptosis.
  JOURNAL   Elife. 2013 Oct 15;2:e00822. doi: 10.7554/eLife.00822.
  PUBMED    24137534
REFERENCE   2  (bases 1 to 8601)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8601
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        2382..2881
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2417..2440)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2814..2833
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2902..3501
                     /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(2902..2921)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3398..3418
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    3578..4130
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3722..3739)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3949..3968
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4142..4861
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4187..4208)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4448..4467)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4795..4816
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4963..5477
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(5646..5662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5646..5662)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5659..5681)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5670..5686
                     /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)."
     promoter        complement(5694..5724)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5739..5760
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5877..5894)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6048..6636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6128..6147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6807..7667)
                     /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     7430..7449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7668..7772)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7840..7858
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7896..7918)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8018..8037
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8231..8253
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8381..8400
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             8600..515
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatctctcga gttgaggtgt taattctaat
     1441 tatctatttc aaatttagca ggaaaaaaga gaacatcacc ttgtaaaact gaagattgtg
     1501 accagtcaga ataatgtcaa agtgcttaca gtgcaggtag tgatatgtgc atctactgca
     1561 gtgaaggcac ttgtagcatt atggtgacag ctgcctcggg aagccaagtt gggctttaaa
     1621 gtgcagggcc tgctgatgtt gagtgctttt tgttctaagg tgcatctagt gcagatagtg
     1681 aagtagatta gcatctactg ccctaagtgc tccttctggc ataagaagtt atgtattcat
     1741 ccaataattc aagccaagca agtatatagg tgttttaata gtttttgttt gcagtcctct
     1801 gttagttttg catagttgca ctacaagaag aatgtagttg tgcaaatcta tgcaaaactg
     1861 atggtggcct gctatttcct tcaaatgaat gatttttact aattttgtgt acttttattg
     1921 tgtcgatgta gaatctgcct ggtctatctg atgtgacagc ttctgtagca ctaaagtgct
     1981 tatagtgcag gtagtgttta gttatctact gcattatgag cacttaaagt actgctagct
     2041 gtagaactcc agcttcggcc tgtcgcccaa tcaaactgtc ctgttactga acactgttct
     2101 atggttagtt ttgcaggttt gcatccagct gtgtgatatt ctgctgtgca aatccatgca
     2161 aaactgactg tggtagtgaa aagtctgtag aaaagtaagg gaaactcaaa cccctttcta
     2221 cacaggttgg gatcggttgc aatgctgtgt ttctgtatgg tattgcactt gtcccggcct
     2281 gttgagtttg gtggggattg tgaccagaag attttgaaaa ttaaatatta ctgaagattt
     2341 cgacttccac tgttaaatgt acaagataca tgaattctac cgggtagggg aggcgctttt
     2401 cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag
     2461 tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt
     2521 tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc
     2581 gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca
     2641 gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca
     2701 gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct
     2761 caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg
     2821 cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc
     2881 gacctgcagc ccaagcttac catgaccgag tacaagccca cggtgcgcct cgccacccgc
     2941 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     3001 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     3061 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     3121 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     3181 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     3241 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     3301 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     3361 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     3421 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     3481 acccgcaagc ccggtgcctg acgcccgccc cacgacccgc agcgcccgac cgaaaggagc
     3541 gcacgacccc atcatccaat tccgcccccc ccccctaacg ttactggccg aagccgcttg
     3601 gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc
     3661 aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc
     3721 cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa
     3781 gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct
     3841 ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca
     3901 caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca
     3961 agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat
     4021 ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc
     4081 cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaac
     4141 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     4201 cggcgacgta aacggccaca agttcagcgt gtctggcgag ggcgagggcg atgccaccta
     4261 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     4321 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     4381 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     4441 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     4501 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     4561 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     4621 cggcatcaag gcgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     4681 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     4741 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     4801 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     4861 atgaattaat taagaattat caagcttatc gataccgtcg acctgcagcc aagcttatcg
     4921 ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
     4981 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaat acataactga
     5041 gaatagagaa gttcagatca aggttaggaa cagagagaca gcagaatatg ggccaaacag
     5101 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat
     5161 gcggtcccgc cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga
     5221 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     5281 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag
     5341 tcctccgata gactgcgtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc
     5401 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt
     5461 cagcgggggt ctttcatggg taacagtttc ttgaagttgg agaacaacat tctgagggta
     5521 ggagtcgaat attaagtaat cctgactcaa ttagccactg ttttgaatcc acatactcca
     5581 atactcctga aatagttcat tatggacagc gcagaagagc tggggagaat taattcgtaa
     5641 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5701 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5761 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5821 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5881 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5941 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6001 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6061 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6121 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6181 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6241 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6301 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6361 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6421 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6481 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6541 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6601 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6661 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6721 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6781 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6841 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6901 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6961 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7021 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7081 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7141 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7201 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7261 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7321 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7381 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7441 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7501 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7561 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7621 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7681 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7741 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7801 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     7861 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     7921 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     7981 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     8041 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     8101 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     8161 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     8221 ccagggtttt cccagtcacg acgttgtaaa acgacggcgc aaggaatggt gcatgcaagg
     8281 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     8341 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     8401 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagaggc
     8461 gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt ttgactcaac
     8521 aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag attttattta
     8581 gtctccagaa aaagggggga a
//
