LOCUS       sequence-115533-        5584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of miR216a in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5584)
  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 5584)
  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-216a
FEATURES             Location/Qualifiers
     source          1..5584
                     /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     1280..1302
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     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             2167..2766
                     /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(2167..2186)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2663..2683
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2865..3458
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3659..3676)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3830..4418)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3910..3929)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4596..5387,5388..5456))
                     /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     5219..5238
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5457..5561)
                     /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 tcctcttcct
     1261 ccatccgccc cgtctctccc ccttgaacct cctcgttcga ccccgcctcg 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 aaaagcttgc gaatccaata aaggcaaatt aattcccaag gctggtttcc tttccagcac
     1801 aacagcactt tagaaaaaga ctttactaag attaaaagag gttgagcatg aaacagtaac
     1861 catcgaaact tttccccccg tcttggtcat gaaatagaaa tttggtattt aattttggtt
     1921 gccatgcaag tcttttgatt cgtgagggct aggaaattgc tctgtttagc ataatcccag
     1981 agaccactgt gagggattgt atgaacatct cacagttgcc agctgagatt aagccaactc
     2041 acagccatcc gtgttagacc tgcaagagaa agtgtatgag cagaggcacg tggctgcctc
     2101 aggtggaagc acatcttcat acatcgagtg caccagcaga agccctgatg tacccctaag
     2161 cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     2221 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     2281 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     2341 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     2401 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     2461 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     2521 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     2581 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     2641 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     2701 accgccgacg tcgagtgccc gaaggaccgc gcgacctggt gcatgacccg caagcccggt
     2761 gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca
     2821 tcgataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc
     2881 tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa
     2941 ctgagaatag agaagttcag atcaaggtca ggaacagatg gaacagctga atatgggcca
     3001 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggaaca
     3061 gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     3121 agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagagaa ccatcagatg
     3181 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     3241 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     3301 cccctcactc ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca
     3361 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
     3421 ctgagtgatt gactacccgt cagcgggggt ctttcatttc cgacttgtgg tctcgctgcc
     3481 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtcttca catgcagcat
     3541 gtatcaaaat taatttggtt ttttttctta agtatttaca ttaaatggcc atagttgcat
     3601 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttggcgctct tccgcttcct
     3661 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3721 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3781 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3841 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3901 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3961 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4021 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4081 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4141 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4201 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4261 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4321 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4381 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4441 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4501 caaaaaggat cttcacctag atccttttaa attaaaaatg aagtttgcgg ccgcaaatca
     4561 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4621 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4681 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4741 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4801 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4861 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4921 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4981 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5041 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5101 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5161 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat
     5221 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5281 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5341 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5401 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5461 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5521 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5581 ccac
//
