LOCUS       Exported                7585 bp ds-DNA     circular SYN 20-DEC-2021
DEFINITION  Empty vector to express overexpression cDNA.
ACCESSION   .
VERSION     .
KEYWORDS    pHAGE_puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7585)
  AUTHORS   Lu B, Klingbeil O, Tarumoto Y, Somerville TDD, Huang YH, Wei Y, Wai 
            DC, Low JKK, Milazzo JP, Wu XS, Cao Z, Yan X, Demerdash OE, Huang G,
            Mackay JP, Kinney JB, Shi J, Vakoc CR
  TITLE     A Transcription Factor Addiction in Leukemia Imposed by the MLL 
            Promoter Sequence.
  JOURNAL   Cancer Cell. 2018 Dec 10;34(6):970-981.e8. doi: 
            10.1016/j.ccell.2018.10.015. Epub 2018 Nov 29.
  PUBMED    30503706
REFERENCE   2  (bases 1 to 7585)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.ccell.2018.10.015"; journalName: "Cancer Cell"; date: 
            "2018-12-10- 10"; volume: "34"; issue: "6"; pages: "970-981.e8"
FEATURES             Location/Qualifiers
     source          1..7585
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     52..71
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             140..739
                     /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(140..159)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     636..656
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    753..1341
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(806..826)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1224..1235)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1416..1649
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(1768..1786)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1854..1958
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1959..2819
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2177..2196)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2990..3578
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3479..3498
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3732..3749
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3841..3860
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     protein_bind    3989..4010
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4025..4055
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4063..4079
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4068..4090
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4087..4103
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4087..4103
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4153..4172
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             4364..4997
                     /label=3 LTR
                     /note="3 long terminal repeat (LTR) from HIV-1"
     misc_feature    5044..5169
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5666..5899
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6084..6128
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    6421..6538
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        6590..6893
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6894..7097
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7048..7068
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7094..7118
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        join(7205..7585,1..119)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7240..7263)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"