LOCUS       sequence-129758-        8049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses miR-146a in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8049)
  AUTHORS   Chang TC, Yu D, Lee YS, Wentzel EA, Arking DE, West KM, Dang CV, 
            Thomas-Tikhonenko A, Mendell JT
  TITLE     Widespread microRNA repression by Myc contributes to tumorigenesis.
  JOURNAL   Nat Genet. 2008 Jan;40(1):43-50. Epub 2007 Dec 9.
  PUBMED    18066065
REFERENCE   2  (bases 1 to 8049)
  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"; journalName: "Nat 
            Genet."; date: "2008-01"; volume: "40(1)"; pages: "43-50. Epub 2007 
            Dec 9"
COMMENT     Sequence Label: pMSCV-PIG-miR-146a
FEATURES             Location/Qualifiers
     source          1..8049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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        1829..2328
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1864..1887)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2261..2280
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2349..2948
                     /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(2349..2368)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2845..2865
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    3009..3576
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3026..3578
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3170..3187)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3397..3416
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3590..3592,3593..3595,3596..4309)
                     /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(3635..3656)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3896..3915)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4243..4264
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4411..4925
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(5094..5110)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5094..5110)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5107..5129)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5118..5134
                     /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(join(5142..5148,5149..5166,5167..5172))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5187..5208
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5325..5342)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5496..6084)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5576..5595)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6255..7046,7047..7115))
                     /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     6878..6897
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7116..7220)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7288..7306
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7344..7366)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7466..7485
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7679..7701
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7829..7848
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8048..8049,1..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 gagagatcca cccacatcag
     1441 ccttccagac tgctggcctg gtctcctcca gatgtttata actcatgagt gccaggacta
     1501 gacctggtac taggaagcag ctgcattgga tttaccaggc ttttcactct tgtattttac
     1561 agggctggga caggcctgga ctgcaaggag gggtctttgc accatctctg aaaagccgat
     1621 gtgtatcctc agctttgaga actgaattcc atgggttgtg tcagtgtcag acctgtgaaa
     1681 ttcagttctt cagctgggat atctctgtca tcgtgggctt gaggacctgg agagagtaga
     1741 tcctgaagaa ctttttcagt ctgctgaaga gcttggaaga ctggagacag aaggcagagt
     1801 ctcaggctcg aggttaacga attctaccgg gtaggggagg cgcttttccc aaggcagtct
     1861 ggagcatgcg ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct
     1921 cgcacacatt ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc
     1981 gcgccacctt ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg
     2041 tgcaggacgt gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc
     2101 gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc
     2161 gctttctggg ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc
     2221 aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag
     2281 cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcagccca
     2341 agcttaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca
     2401 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
     2461 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
     2521 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
     2581 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
     2641 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
     2701 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
     2761 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
     2821 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
     2881 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
     2941 gtgcctgacg cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg
     3001 catcgaattc cgcccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg
     3061 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc
     3121 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa
     3181 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga
     3241 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc
     3301 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc
     3361 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac
     3421 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg
     3481 tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac
     3541 ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaacca tggtgagcaa
     3601 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     3661 cggccacaag ttcagcgtgt ctggcgaggg cgagggcgat gccacctacg gcaagctgac
     3721 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     3781 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     3841 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     3901 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     3961 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     4021 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggc
     4081 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     4141 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     4201 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     4261 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat gaattaatta
     4321 agaattatca agcttatcga taccgtcgac ctgcagccaa gcttatcgat aaaataaaag
     4381 attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag
     4441 ctagcttaag taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt
     4501 tcagatcaag gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt
     4561 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtcccgccc
     4621 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     4681 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     4741 tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgataga
     4801 ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
     4861 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
     4921 ttcatgggta acagtttctt gaagttggag aacaacattc tgagggtagg agtcgaatat
     4981 taagtaatcc tgactcaatt agccactgtt ttgaatccac atactccaat actcctgaaa
     5041 tagttcatta tggacagcgc agaagagctg gggagaatta attcgtaatc atggtcatag
     5101 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5161 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5221 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5281 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5341 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5401 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5461 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5521 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5581 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5641 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5701 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5761 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5821 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5881 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     5941 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6001 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6061 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6121 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6181 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6241 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6301 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6361 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6421 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6481 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6541 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6601 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6661 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6721 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6781 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6841 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6901 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6961 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7021 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7081 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7141 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7201 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     7261 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     7321 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     7381 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     7441 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     7501 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
     7561 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     7621 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     7681 cagtcacgac gttgtaaaac gacggcgcaa ggaatggtgc atgcaaggag atggcgccca
     7741 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     7801 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg
     7861 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggcga ttagtccaat
     7921 ttgttaaaga caggatatca gtggtccagg ctctagtttt gactcaacaa tatcaccagc
     7981 tgaagcctat agagtacgag ccatagataa aataaaagat tttatttagt ctccagaaaa
     8041 aggggggaa
//
