LOCUS       40924_11700        6219 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector for expressing microRNA (miRNA) precursors using the CMV 
            promoter, with a GFP reporter for transfection and a neomycin 
            resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6219)
  AUTHORS   OriGene
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6219)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Inserts are cloned between the SgfI (AsiSI) and MluI sites.
FEATURES             Location/Qualifiers
     source          1..6219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1084..1113
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     CDS             1132..1155
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     primer_bind     complement(1175..1191)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1202..1824
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     rep_origin      complement(1973..2561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    complement(2890..2937)
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(3162..3857)
                     /codon_start=1
                     /label=TurboGFP
                     /note="improved green fluorescent protein from Pontellina 
                     plumata (Evdokimov et al., 2006)"
                     /translation="MESDESGLPAMEIECRITGTLNGVEFELVGGGEGTPEQGRMTNKM
                     KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL
                     HVSFSYRYEAGRVIGDFKVMGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNDLDGSFTR
                     TFSLRDGGYYSSVVDSHMHFKSAIHPSILQNGGPMFAFRRVEEDHSNTELGIVEYQHAF
                     KTPDADAGEE"
     misc_feature    complement(3858..4437)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             complement(4462..5253)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(5288..5645)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        complement(5647..5751)
                     /label=AmpR promoter
     rep_origin      5778..6206
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccagt accgaggaga tctgcgccgc
     1021 gatcgccggc gcgccagatc tcaagcttaa ctagctagcg gaccgacgcg tacgcggccg
     1081 ctcgagcaga aactcatctc agaagaggat ctggcagcaa atgatatcct ggattacaag
     1141 gatgacgacg ataaggttta aacggccggc cgcggtcata gctgtttcct gaacagatcc
     1201 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     1261 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     1321 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     1381 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     1441 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     1501 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     1561 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     1621 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     1681 gattttaaaa taactatacc agcaggagga cgtccagaca cagcataggc tacctggcca
     1741 tgcccaaccg gtgggacatt tgagttgctt gcttggcact gtcctctcat gcgttgggtc
     1801 cactcagtag atgcctgttg aattgggtac gcggccagcg gcgagcggta tcagctcact
     1861 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1921 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1981 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2041 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2101 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2161 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2221 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2281 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2341 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2401 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2461 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2521 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2581 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2641 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2701 aaagtatata tgagtaacct gaggctatgg cagggcctgc cgccccgacg ttggctgcga
     2761 gccctgggcc ttcacccgaa cttggggggt ggggtgggga aaaggaagaa acgcgggcgt
     2821 attggcccca atggggtctc ggtggggtat cgacagagtg ccagccctgg gaccgaaccc
     2881 cgcgtttatg aacaaacgac ccaacaccgt gcgttttatt ctgtcttttt attgccgtca
     2941 tagcgcgggt tccttccggt attgtctcct tccgtgtttc agttagcctc cccctagggt
     3001 gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga
     3061 aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg
     3121 gcaggttggg cgtcgcttgg tcggtcattt tcttcgaatt attcttcacc ggcatctgca
     3181 tccggggtct tgaaggcgtg ctggtactcc acgatgccca gctcggtgtt gctgtgatcc
     3241 tcctccacgc ggcggaaggc gaacatgggg cccccgttct gcaggatgct ggggtggatg
     3301 gcgctcttga agtgcatgtg gctgtccacc acggagctgt agtagccgcc gtcgcgcagg
     3361 ctgaaggtgc gggtgaagct gccatccaga tcgttatcgc ccatggggtg caggtgctcc
     3421 acggtggcgt tgctgcggat gatcttgtcg gtgaagatca cgctgtcctc ggggaagccg
     3481 gtgcccatca ccttgaagtc gccgatcacg cggccggcct cgtagcggta gctgaagctc
     3541 acgtgcagca cgccgccgtc ctcgtacttc tcgatgcggg tgttggtgta gccgccgttg
     3601 ttgatggcgt gcaggaaggg gttctcgtag ccgctggggt aggtgccgaa gtggtagaag
     3661 ccgtagccca tcacgtggct cagcaggtag gggctgaagg tcagggcgcc tttggtgctc
     3721 ttcatcttgt tggtcatgcg gccctgctcg ggggtgccct ctccgccgcc caccagctcg
     3781 aactccacgc cgttcagggt gccggtgatg cggcactcga tctccatggc gggcaggccg
     3841 ctctcgtcgc tctccatggt tgtggccata ttatcatcgt gtttttcaaa ggaaaaccac
     3901 gtccccgtgg ttcggggggc ctagacgttt ttttaacctc gactaaacac atgtaaagca
     3961 tgtgcaccga ggccccagat cagatcccat acaatggggt accttctggg catccttcag
     4021 ccccttgttg aatacgcttg aggagagcca tttgactctt tccacaacta tccaactcac
     4081 aacgtggcac tggggttgtg ccgcctttgc aggtgtatct tatacacgtg gcttttggcc
     4141 gcagaggcac ctgtcgccag gtggggggtt ccgctgcctg caaagggtcg ctacagacgt
     4201 tgtttgtctt caagaagctt ccagaggaac tgcttccttc acgacattca acagaccttg
     4261 cattcctttg gcgagagggg aaagacccct aggaatgctc gtcaagaaga cagggccagg
     4321 tttccgggcc ctcacattgc caaaagacgg caatatggtg gaaaataaca tatagacaaa
     4381 cgcacaccgg ccttattcca agcggcttcg gccagtaacg ttaggggggg gggcggaatt
     4441 cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg
     4501 cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag
     4561 ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcgatccg ccacacccag
     4621 ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca
     4681 ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgctcgcct tgagcctggc
     4741 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
     4801 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
     4861 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
     4921 ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag
     4981 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
     5041 ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc
     5101 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
     5161 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
     5221 agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg
     5281 atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     5341 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     5401 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     5461 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     5521 actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     5581 gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc
     5641 ctcaggactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5701 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5761 ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5821 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5881 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5941 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     6001 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     6061 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     6121 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     6181 gatttaacaa aaatttaacg cgaattttaa caaaatatt
//
