LOCUS       sequence-109974-        5510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of miR148a with scrambled seed in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5510)
  AUTHORS   Bofill-De Ros X, Gironella M, Fillat C
  TITLE     miR-148a- and miR-216a-regulated oncolytic adenoviruses targeting 
            pancreatic tumors attenuate tissue damage without perturbation of 
            miRNA activity.
  JOURNAL   Mol Ther. 2014 Sep;22(9):1665-77. doi: 10.1038/mt.2014.98. Epub 2014
            Jun 4.
  PUBMED    24895996
REFERENCE   2  (bases 1 to 5510)
  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: 
            "10.1038/mt.2014.98"; journalName: "Mol Ther"; date: "2014-09"; 
            volume: "22"; issue: "9"; pages: "1665-77"
COMMENT     Sequence Label: p-miR-SC
FEATURES             Location/Qualifiers
     source          1..5510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             934..1350
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1318..1334
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(1408..1428)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1413..1742
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1593..1728
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2093..2692
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(2093..2112)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2589..2609
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2791..3384
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3585..3602)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3756..4344)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3836..3855)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4522..5313,5314..5382))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5145..5164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5383..5487)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt
      901 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt
      961 ttgaccttag atcactggaa agatgtcgag cggctcgctc acaaccagtc ggtagatgtc
     1021 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg
     1081 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca
     1141 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct
     1201 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttctt
     1261 ccatccgcgc cgtctctccc ccttgaacct cctctttcga ccccgcctca atcctccctt
     1321 tatccagccc tcactccttc tctaggcgcc ggccggatcc cagtgtggtg gtacgtagga
     1381 attcgccagc acagtggtcg accctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     1441 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     1501 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     1561 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     1621 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     1681 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     1741 aaaagcttgt tccattatcg gtcgcatcct gaactaaatt gcttttaacg agttattctt
     1801 ctttgtctta gctggggggt gggggaacac acctgcagga agaaacttct ggaggaagac
     1861 agcacgtttg gtcttttgag gcaaagttct gagacactcc gactctgagt atgatagaag
     1921 ttgcacgcct acagaacttt gtctctagag gctgtggtcg ccgatgccgc cagtgctcca
     1981 gacgccaggc cgagtggccc tggtggccac gtcgctgcgc ctttcccttg ggggaggggc
     2041 aaggccagag ggggtccaag tgcagcacga ggaacttgac caaaagctta ccatgaccga
     2101 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     2161 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     2221 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     2281 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     2341 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     2401 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     2461 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     2521 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     2581 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     2641 gtgcccgaag gaccgcgcga cctggtgcat gacccgcaag cccggtgcct gacgcccgcc
     2701 ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcga taaaataaaa
     2761 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     2821 gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa
     2881 gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt
     2941 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc
     3001 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     3061 ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     3121 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     3181 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg
     3241 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
     3301 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     3361 acccgtcagc gggggtcttt catttccgac ttgtggtctc gctgccttgg gagggtctcc
     3421 tctgagtgat tgactacccg tcagcggggg tcttcacatg cagcatgtat caaaattaat
     3481 ttggtttttt ttcttaagta tttacattaa atggccatag ttgcattaat gaatcggcca
     3541 acgcgcgggg agaggcggtt tgcgtattgg cgctcttccg cttcctcgct cactgactcg
     3601 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3661 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3721 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3781 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3841 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3901 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3961 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4021 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4081 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4141 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     4201 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4261 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4321 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4381 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4441 acctagatcc ttttaaatta aaaatgaagt ttgcggccgc aaatcaatct aaagtatata
     4501 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4561 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4621 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4681 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4741 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4801 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4861 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4921 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4981 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5041 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5101 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca
     5161 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5221 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5281 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5341 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5401 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5461 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
//
