LOCUS       Exported                6340 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pMKO.1 puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6340)
  AUTHORS   Stewart SA, Dykxhoorn DM, Palliser D, Mizuno H, Yu EY, An DS, 
            Sabatini DM, Chen IS, Hahn WC, Sharp PA, Weinberg RA, Novina CD
  TITLE     Lentivirus-delivered stable gene silencing by RNAi in primary cells.
  JOURNAL   RNA 2003 Apr;9(4):493-501.
  PUBMED    12649500
REFERENCE   2  (bases 1 to 6340)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "RNA"; date:
            "2003-04"; volume: "9(4)"; pages: "493-501"
FEATURES             Location/Qualifiers
     source          1..6340
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..241
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5
                     /note="Human U6 promoter, forward primer"
     promoter        310..639
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      490..625
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             649..1248
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(649..668)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1145..1165
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1444..1869
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(1972..1994)
                     /label=pGEX 3
                     /note="pGEX vectors, reverse primer"
     promoter        2135..2464
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2315..2450
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2486..2505
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2586..2603)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2757..3345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2837..2856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3516..4376)
                     /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     4139..4158
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4377..4481)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4549..4567
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        4667..5046
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5047..5250
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5201..5221
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5251..5426
                     /label=5 LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5489..5688
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5889..6305
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6235..6257
                     /label=pLXSN 5
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6273..6289
                     /label=pBABE 5
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"