LOCUS       Exported                6492 bp ds-DNA     circular SYN 06-OCT-2021
DEFINITION  Cloning vector pmirGLO, complete sequence.
ACCESSION   FJ376737
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6492)
  AUTHORS   Garvin D.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-OCT-2008) Scientific Communication, Promega 
            Corporation, 2800 Woods Hollow Rd., Madison, WI 53711, USA
REFERENCE   2  (bases 1 to 6492)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Oct 7, 2021 from SnapGene 5.2.4
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (10-OCT-2008) Scientific Communication, Promega Corporation, 2800 
            Woods Hollow Rd., Madison, WI 53711, USA"
FEATURES             Location/Qualifiers
     source          1..6492
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          90..6492
                     /organism="Cloning vector pmirGLO"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
                     /db_xref="taxon:574161"
     primer_bind     89..107
                     /label=+1fs Fwd
                     /note="color: black; sequence: gaattccGCTTCCAAGGTGTACGAC; 
                     added: 2021-03-03"
     primer_bind     89..107
                     /label=-1fs Fwd
                     /note="color: black; sequence: gaattcccGCTTCCAAGGTGTACGAC; 
                     added: 2021-03-03"
     CDS             90..1862
                     /codon_start=1
                     /product="hRluc-neo fusion protein"
                     /label=hRluc-neo fusion protein
                     /note="Renilla luciferase-neomycin resistance 
                     fusionprotein; used as a control reporter for 
                     normalization"
                     /note="color: #993366"
                     /protein_id="ACJ37464.1"
                     /translation="ASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAENA
                     VIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAWF
                     ELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDIA
                     LIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPREI
                     PLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKVK
                     GLHFSQEDAPDEMGKYIKSFVERVLKNEQTGGGSGGGGSGGGGSGGIEQDGLHAGSPAA
                     WVERLFGYDWAQQTIGCSDAAVFRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLATT
                     GVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCP
                     FDHQAKHRIERARTRMEAGLVDQDDLDEEHQGLAPAELFARLKARMPDGEDLVVTHGDA
                     CLPNIMVENGRFSGFIDCGRLGVADRYQDIALATRDIAEELGGEWADRFLVLYGIAAPD
                     SQRIAFYRLLDEFF"
     CDS             91..93
                     /codon_start=1
                     /label=+1 frame
                     /note="color: #993366"
                     /translation="L"
     CDS             92..94
                     /codon_start=1
                     /label=+2 frame
                     /note="color: #993366"
                     /translation="F"
     primer_bind     complement(110..137)
                     /label=Primer 1
                     /note="color: black; sequence: 
                     TCCCACTCCTTTCAAGACCTnnnnnnnnnnatgcgtttgcgttgctcg; added: 
                     2020-07-12"
     primer_bind     complement(176..204)
                     /label=RLuc-SeqR
                     /note="color: black; sequence: 
                     cggaatcatagtagttgatgaaggagtcc; added: 2020-04-19"
     regulatory      1926..1974
                     /regulatory_class="polyA_signal_sequence"
                     /label=synthetic polyadenylation signal
                     /note="synthetic polyadenylation signal"
                     /note="color: #ffffff"
     CDS             2235..3095
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="confers resistance to ampicillin and is used 
                     aselectable marker in E. coli"
                     /note="color: #993366"
                     /protein_id="ACJ37465.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3250..3286
                     /label=ColE1-derived plasmid origin of replication
                     /note="ColE1-derived plasmid origin of replication"
                     /note="color: #ffff00"
     regulatory      4292..4807
                     /regulatory_class="promoter"
                     /label=human phosphoglycerate kinase promoter
                     /note="human phosphoglycerate kinase promoter"
                     /note="color: #ffffff"
     CDS             4843..6492
                     /codon_start=1
                     /product="modified firefly luciferase"
                     /label=modified firefly luciferase
                     /note="luc2; reporter of miRNA activity in mammaliancells"
                     /note="color: #993366"
                     /protein_id="ACJ37466.1"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
ORIGIN
        1 aactcgtttg ggtttgcggt gtaagtgcag cccgtcttac accgtgcggc acaggcacta
       61 gtactgatgt cgtatacagg gcttttgacg cttccaaggt gtacgacccc gagcaacgca
      121 aacgcatgat cactgggcct cagtggtggg ctcgctgcaa gcaaatgaac gtgctggact
      181 ccttcatcaa ctactatgat tccgagaagc acgccgagaa cgccgtgatt tttctgcatg
      241 gtaacgctgc ctccagctac ctgtggaggc acgtcgtgcc tcacatcgag cccgtggcta
      301 gatgcatcat ccctgatctg atcggaatgg gtaagtccgg caagagcggg aatggctcat
      361 atcgcctcct ggatcactac aagtacctca ccgcttggtt cgagctgctg aaccttccaa
      421 agaaaatcat ctttgtgggc cacgactggg gggcttgtct ggcctttcac tactcctacg
      481 agcaccaaga caagatcaag gccatcgtcc atgctgagag tgtcgtggac gtgatcgagt
      541 cctgggacga gtggcctgac atcgaggagg atatcgccct gatcaagagc gaagagggcg
      601 agaaaatggt gcttgagaat aacttcttcg tcgagaccat gctcccaagc aagatcatgc
      661 ggaaactgga gcctgaggag ttcgctgcct acctggagcc attcaaggag aagggcgagg
      721 ttagacggcc taccctctcc tggcctcgcg agatccctct cgttaaggga ggcaagcccg
      781 acgtcgtcca gattgtccgc aactacaacg cctaccttcg ggccagcgac gatctgccta
      841 agatgttcat cgagtccgac cctgggttct tttccaacgc tattgtcgag ggagctaaga
      901 agttccctaa caccgagttc gtgaaggtga agggcctcca cttcagccag gaggacgctc
      961 cagatgaaat gggtaagtac atcaagagct tcgtggagcg cgtgctgaag aacgagcaga
     1021 ccggtggtgg gagcggaggt ggcggatcag gtggcggagg ctccggaggg attgaacaag
     1081 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     1141 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     1201 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     1261 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     1321 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     1381 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     1441 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     1501 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     1561 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     1621 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     1681 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     1741 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     1801 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     1861 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgatg
     1921 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga
     1981 tagcgataag gatcctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga
     2041 cattcatccg gggtcagcac cgtttctgcg gactggcttt ctacgtaatg gtttcttaga
     2101 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2161 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2221 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2281 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2341 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2401 agagttttcg ccccgaagaa cgttttccaa tgatgagcac tttcaaagtt ctgctatgtg
     2461 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2521 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2581 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2641 ttctgacaac tatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2701 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2761 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     2821 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     2881 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2941 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3001 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3061 ctgagatagg tgcctcactg attaagcatt ggtaattcga aatgaccgac caagcgacgc
     3121 ccaaccggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3181 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3241 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3301 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3361 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3421 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3481 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3541 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3601 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3661 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     3721 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3781 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatttca
     3841 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3901 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tatagtccgg
     3961 aaatacagga acgcacgctg gatggccctt cgctgggatg gtgaaaccat gaaaaatggc
     4021 agcttcagtg gattaagtgg gggtaatgtg gcctgtaccc tctggttgca taggtattca
     4081 tacggttaaa atttatcagg cgcgattgcg gcagtttttc gggtggtttg ttgccatttt
     4141 tacctgtctg ctgccgtgat cgcgctgaac gcgttttagc ggtgcgtaca attaagggat
     4201 tatggtaaat ccacttactg tctgccctcg tagccatcga gataaaccgc agtactccgg
     4261 ccacgatgcg tccggcgtag aggatcgaga tctaccgggt aggggaggcg cttttcccaa
     4321 ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc
     4381 tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg
     4441 gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc
     4501 tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg
     4561 acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt
     4621 gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg
     4681 gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc
     4741 ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct
     4801 gcagcccaag cttggcaatc cggtactgtt ggtaaagcca ccatggaaga tgccaaaaac
     4861 attaagaagg gcccagcgcc attctaccca ctcgaagacg ggaccgccgg cgagcagctg
     4921 cacaaagcca tgaagcgcta cgccctggtg cccggcacca tcgcctttac cgacgcacat
     4981 atcgaggtgg acattaccta cgccgagtac ttcgagatga gcgttcggct ggcagaagct
     5041 atgaagcgct atgggctgaa tacaaaccat cggatcgtgg tgtgcagcga gaatagcttg
     5101 cagttcttca tgcccgtgtt gggtgccctg ttcatcggtg tggctgtggc cccagctaac
     5161 gacatctaca acgagcgcga gctgctgaac agcatgggca tcagccagcc caccgtcgta
     5221 ttcgtgagca agaaagggct gcaaaagatc ctcaacgtgc aaaagaagct accgatcata
     5281 caaaagatca tcatcatgga tagcaagacc gactaccagg gcttccaaag catgtacacc
     5341 ttcgtgactt cccatttgcc acccggcttc aacgagtacg acttcgtgcc cgagagcttc
     5401 gaccgggaca aaaccatcgc cctgatcatg aacagtagtg gcagtaccgg attgcccaag
     5461 ggcgtagccc taccgcaccg caccgcttgt gtccgattca gtcatgcccg cgaccccatc
     5521 ttcggcaacc agatcatccc cgacaccgct atcctcagcg tggtgccatt tcaccacggc
     5581 ttcggcatgt tcaccacgct gggctacttg atctgcggct ttcgggtcgt gctcatgtac
     5641 cgcttcgagg aggagctatt cttgcgcagc ttgcaagact ataagattca atctgccctg
     5701 ctggtgccca cactatttag cttcttcgct aagagcactc tcatcgacaa gtacgaccta
     5761 agcaacttgc acgagatcgc cagcggcggg gcgccgctca gcaaggaggt aggtgaggcc
     5821 gtggccaaac gcttccacct accaggcatc cgccagggct acggcctgac agaaacaacc
     5881 agcgccattc tgatcacccc cgaaggggac gacaagcctg gcgcagtagg caaggtggtg
     5941 cccttcttcg aggctaaggt ggtggacttg gacaccggta agacactggg tgtgaaccag
     6001 cgcggcgagc tgtgcgtccg tggccccatg atcatgagcg gctacgttaa caaccccgag
     6061 gctacaaacg ctctcatcga caaggacggc tggctgcaca gcggcgacat cgcctactgg
     6121 gacgaggacg agcacttctt catcgtggac cggctgaaga gcctgatcaa atacaagggc
     6181 taccaggtag ccccagccga actggagagc atcctgctgc aacaccccaa catcttcgac
     6241 gccggggtcg ccggcctgcc cgacgacgat gccggcgagc tgcccgccgc agtcgtcgtg
     6301 ctggaacacg gtaaaaccat gaccgagaag gagatcgtgg actatgtggc cagccaggtt
     6361 acaaccgcca agaagctgcg cggtggtgtt gtgttcgtgg acgaggtgcc taaaggactg
     6421 accggcaagt tggacgcccg caagatccgc gagattctca ttaaggccaa gaagggcggc
     6481 aagatcgccg tg
//
