LOCUS       Exported                6740 bp ds-DNA    circular SYN 26-2-2016
DEFINITION  Retroviral vector with an mAmetrine marker, for expressing a short 
            hairpin RNA (shRNA) with an ""UltramiR"" microRNA scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    pLMPd-Ametrine
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6740)
  AUTHORS   transOMIC
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6740
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
     misc_feature    579..920
                     /note="MESV Psi"
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /note="gag (truncated)"
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     ncRNA           1443..1564
                     /note="5 miR-30a"
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     misc_RNA        1565..1633
                     /note="shRNA"
                     /note="short hairpin RNA"
     gap             1568..1589
                     /estimated_length="22"
     gap             1609..1630
                     /estimated_length="22"
     ncRNA           1634..1760
                     /note="3 miR-30a"
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     promoter        1774..2273
                     /note="PGK promoter"
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             2294..3013
                     /codon_start=1
                     /product="photostable yellow fluorescent protein with a 
                     large Stokes shift, excitable with violet light (Ai et al.,
                     2008)"
                     /note="mAmetrine1.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVRGEGEGDATNGKLTL
                     KFICTSGELPVPWPTLVTTLSYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GSYRTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNMNAWDAYITADKQKNGIK
                     ANFKIEHNVKDGGVQLADAYQQNTPIGDGSVLLPDNHYLSFQSKLFKDPNEQRDHMVLL
                     EFVTAAGITPGMDELYK"
     LTR             3102..3616
                     /note="3 LTR"
                     /note="3 long terminal repeat from murine embryonic stem 
                     cell virus"
     rep_origin      complement(4187..4775)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4946..5806)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5807..5911)
                     /gene="bla"
                     /note="AmpR promoter"
     LTR             6739..515
                     /note="5 LTR"
                     /note="5 long terminal repeat from murine embryonic stem 
                     cell virus"