LOCUS       Exported               12020 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  DDX4(1-236) fused to mCh-Cry2WT.
ACCESSION   .
VERSION     .
KEYWORDS    pHR-DDX4N-mCh-Cry2WT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12020)
  AUTHORS   Shin Y, Berry J, Pannucci N, Haataja MP, Toettcher JE, Brangwynne CP
  TITLE     Spatiotemporal Control of Intracellular Phase Transitions Using 
            Light-Activated optoDroplets.
  JOURNAL   Cell. 2017 Jan 12;168(1-2):159-171.e14. doi: 
            10.1016/j.cell.2016.11.054. Epub 2016 Dec 29.
  PUBMED    28041848
REFERENCE   2  (bases 1 to 12020)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell."; 
            date: "2017-01-12"; volume: "168(1-2):159-171.e14. doi"; pages: " 
            10.1016/j.cell.2016.11.054. Epub 2016 Dec 29"
FEATURES             Location/Qualifiers
     source          1..12020
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    43..160
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        313..720
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             2232..3725
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     misc_feature    3877..4465
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3930..3950)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4348..4359)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4468..4484)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4469..4485)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             4575..4808
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(4833..4851)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4834..4851)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5141..5160)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5374..5393)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5493..5515
                     /label=pGEX 3
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5553..5571)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5639..5743
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5744..6604
                     /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(5962..5981)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6775..7363
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7264..7283
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7517..7534
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(7604..7624)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7609..7938
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7789..7924
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7851..7870
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     intron          9072..9137
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             9267..9287
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    9712..9846
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9749..9768)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9803..9822
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             10014..10647
                     /label=3 LTR
                     /note="3 long terminal repeat (LTR) from HIV-1"
     misc_feature    10694..10819
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11316..11549
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11734..11778
                     /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"