LOCUS       sequence-339675-        5014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a positive control reporter with 3' UTR sequences perfectly 
            complementary to miR-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5014)
  AUTHORS   Li P, Chen Y, Juma CA, Yang C, Huang J, Zhang X, Zeng Y
  TITLE     Differential Inhibition of Target Gene Expression by Human 
            microRNAs.
  JOURNAL   Cells. 2019 Jul 30;8(8). pii: cells8080791. doi: 
            10.3390/cells8080791.
  PUBMED    31366019
REFERENCE   2  (bases 1 to 5014)
  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: "Cells."; 
            date: "2019-07-30"; volume: "8(8). pii: cells8080791. doi"; pages: "
            10.3390/cells8080791"
COMMENT     Sequence Label: pCMV-luc-miR-1
FEATURES             Location/Qualifiers
     source          1..5014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     718..737
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1481..1860
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1865..2068
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2019..2039
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2138..3790
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(2204..2222)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(4145..4163)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4231..4335
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4336..4404,4405..5014,1..182)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4554..4573)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
       61 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
      121 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
      181 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
      241 ttaaaaggat ctaggctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
      301 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
      361 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
      421 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
      481 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
      541 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
      601 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
      661 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
      721 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
      781 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg gagatgcgcc
      841 gcgtgcggct gctggagatg gcggacgcga tggatatgtt ctgccaaggg ttggtttgcg
      901 cattcacagt tctccgcaag aattgattgg ctccaattct tggagtggtg aatccgttag
      961 cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa
     1021 cgcggggagg cagacaaggt atagggcggc gcctacaatc catgccaacc cgttccatgt
     1081 gctcgccgag gcggcataaa tcgccgtgac gatcagcggt ccaatgatcg aagttaggct
     1141 ggtaagagcc gcgagcgatc cttgaagctg tccctgatgg tcgtcatcta cctgcctgga
     1201 cagcatggcc tgcaacgcgg gcatcccgat gccgccggaa gcgagaagaa tcataatggg
     1261 gaaggccatc cagcctcgcg tcggggagct ttttgcaaaa gcctaggcct ccaaaaaagc
     1321 ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa
     1381 aaaaaattag tcagccatga gcttggccca ttgcatacgt tgtatccata tcataatatg
     1441 tacatttata ttggctcatg tccaacatta ccgccatgtt gacattgatt attgactagt
     1501 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     1561 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     1621 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     1681 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     1741 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     1801 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     1861 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     1921 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     1981 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     2041 acggtgggag gtctatataa gcagagctcg tttagtgaag tcaagcttaa gtgaccagct
     2101 acagtcggaa accatcagca agaggtcatt gttcaccatg gaagacgcca aaaacataaa
     2161 gaaaggcccg gcgccattct atcctctaga ggatggaacc gctggagagc aactgcataa
     2221 ggctatgaag agatacgccc tggttcctgg aacaattgct tttacagatg cacatatcga
     2281 ggtgaacatc acgtacgcgg aatacttcga aatgtccgtt cggttggcag aagctatgaa
     2341 acgatatggg ctgaatacaa atcacagaat cgtcgtatgc agtgaaaact ctcttcaatt
     2401 ctttatgccg gtgttgggcg cgttatttat cggagttgca gttgcgcccg cgaacgacat
     2461 ttataatgaa cgtgaattgc tcaacagtat gaacatttcg cagcctaccg tagtgtttgt
     2521 ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa aaattaccaa taatccagaa
     2581 aattattatc atggattcta aaacggatta ccagggattt cagtcgatgt acacgttcgt
     2641 cacatctcat ctacctcccg gttttaatga atacgatttt gtaccagagt cctttgatcg
     2701 tgacaaaaca attgcactga taatgaattc ctctggatct actgggttac ctaagggtgt
     2761 ggcccttccg catagaactg cctgcgtcag attctcgcat gccagagatc ctatttttgg
     2821 caatcaaatc attccggata ctgcgatttt aagtgttgtt ccattccatc acggttttgg
     2881 aatgtttact acactcggat atttgatatg tggatttcga gtcgtcttaa tgtatagatt
     2941 tgaagaagag ctgtttttac gatcccttca ggattacaaa attcaaagtg cgttgctagt
     3001 accaacccta ttttcattct tcgccaaaag cactctgatt gacaaatacg atttatctaa
     3061 tttacacgaa attgcttctg ggggcgcacc tctttcgaaa gaagtcgggg aagcggttgc
     3121 aaaacgcttc catcttccag ggatacgaca aggatatggg ctcactgaga ctacatcagc
     3181 tattctgatt acacccgagg gggatgataa accgggcgcg gtcggtaaag ttgttccatt
     3241 ttttgaagcg aaggttgtgg atctggatac cgggaaaacg ctgggcgtta atcagagagg
     3301 cgaattatgt gtcagaggac ctatgattat gtccggttat gtaaacaatc cggaagcgac
     3361 caacgccttg attgacaagg atggatggct acattctgga gacatagctt actgggacga
     3421 agacgaacac ttcttcatag ttgaccgctt gaagtcttta attaaataca aaggatatca
     3481 ggtggccccc gctgaattgg aatcgatatt gttacaacac cccaacatct tcgacgcggg
     3541 cgtggcaggt cttcccgacg atgacgccgg tgaacttccc gccgccgttg ttgttttgga
     3601 gcacggaaag acgatgacgg aaaaagagat cgtggattac gtcgccagtc aagtaacaac
     3661 cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa gtaccgaaag gtcttaccgg
     3721 aaaactcgac gcaagaaaaa tcagagagat cctcataaag gccaagaagg gcggaaagtc
     3781 caaattgtaa ctctagacaa gaggtcattg ttcaccatgg gttgatccgc tagcatacat
     3841 acttctttac attccatact cgagtgacct tcagaccttg gcactggagg tggcccggca
     3901 gaagcgcggc atcgtggatc agtgctgcac cagcatctgc tctctctacc aactggagaa
     3961 ctactgcaac taggcccacc actaccctgt ccacccctct gcaatgaata aaacctttga
     4021 aagagcacta caagttgtgt gtacatgcgt gcatgtgcat atgtggtgcg gggggaacat
     4081 gagtggggct ggctggagtg gcgatgataa gctgtcaaac atgagaattc ttgaagacga
     4141 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     4201 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4261 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4321 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4381 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4441 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4501 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4561 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     4621 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4681 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4741 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4801 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     4861 cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa
     4921 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     4981 ggaccacttc tgcgctcggc ccttccggct ggct
//
