LOCUS       Exported                7084 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    shYAP1 # 1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7084)
  AUTHORS   Rosenbluh J, Nijhawan D, Cox AG, Li X, Neal JT, Schafer EJ, Zack TI,
            Wang X, Tsherniak A, Schinzel AC, Shao DD, Schumacher SE, Weir BA, 
            Vazquez F, Cowley GS, Root DE, Mesirov JP, Beroukhim R, Kuo CJ, 
            Goessling W, Hahn WC
  TITLE     beta-Catenin-Driven Cancers Require a YAP1 Transcriptional Complex 
            for Survival and Tumorigenesis.
  JOURNAL   Cell. 2012 Dec 21;151(7):1457-73. doi: 10.1016/j.cell.2012.11.026. 
            Epub 2012 Dec 13.
  PUBMED    23245941
REFERENCE   2  (bases 1 to 7084)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2012.11.026"; journalName: "Cell"; date: 
            "2012-12-21- 21"; volume: "151"; issue: "7"; pages: "1457-73"
FEATURES             Location/Qualifiers
     source          1..7084
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    1..17
                     /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     6..28
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     25..41
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     25..41
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     60..80
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        62..80
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        108..334
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             335..515
                     /label=5 LTR (truncated)
                     /note="truncated 5 long terminal repeat (LTR) from HIV-1"
     misc_feature    562..687
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1180..1413
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1598..1642
                     /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"
     promoter        1940..2180
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     1940..1960
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2111..2130
                     /label=LKO.1 5
                     /note="Human U6 promoter, forward primer"
     misc_feature    2285..2402
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2451..2961
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2880..2898
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2983..3582
                     /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(2983..3002)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3479..3499
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3710..3943
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4015..4136
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4052..4071)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4106..4125
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4176..4311
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4238..4257
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(4331..4350)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4332..4350)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4360..4377)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4360..4376)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4369..4391)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4518..4973
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4605..4624)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4815..4836
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4999..5103
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5104..5964
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5322..5341)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6135..6723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6624..6643
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6877..6894
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7011..7032
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7047..7077
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"