LOCUS       Exported                6726 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTVDll42AdVenus, complete sequence.
ACCESSION   KF293660
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6726)
  AUTHORS   Ubezio B, Blanco RA, Geudens I, Stanchi F, Mathivet T, Jones ML, 
            Ragab A, Bentley K, Gerhardt H.
  TITLE     Synchronization of endothelial Dll4-Notch dynamics switch blood 
            vessels from branching to expansion
  JOURNAL   eLife 5, e12167 (2016)
  PUBMED    27074663
REFERENCE   2  (bases 1 to 6726)
  AUTHORS   Ubezio B.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JUN-2013) Vascular Biology Laboratory, Cancer Research
            UK, 44 Lincolns Inn Fields, London, London WC2A 3LY, United Kingdom
REFERENCE   3  (bases 1 to 6726)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6726
                     /organism="Cloning vector pTVDll42AdVenus"
                     /mol_type="other DNA"
                     /db_xref="taxon:1501982"
     promoter        471..575
                     /gene="bla"
                     /label=AmpR promoter
     CDS             576..1436
                     /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"
     rep_origin      1607..2195
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2483..2504
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2519..2549
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2557..2573
                     /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     2581..2597
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2618..2636
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     gene            2654..3128
                     /gene="Dll4"
                     /label=Dll4
     intron          2654..3124
                     /gene="Dll4"
                     /number=10
     exon            3125..3128
                     /gene="Dll4"
                     /number=11
     misc_feature    3128..3214
                     /label=P2A sequence
                     /note="P2A sequence"
     CDS             3152..3208
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3215..4093
                     /codon_start=1
                     /product="dVenus"
                     /label=dVenus
                     /protein_id="AIA24530.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYPPKKKRKVEDPLYKSHGFPPAVAAQDDGTLPMSCAQESGMDRH
                     PAACAPARINV"
     CDS             3215..3928
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     CDS             3932..3952
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(4160..4193)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      4251..4766
                     /regulatory_class="promoter"
                     /note="PGK"
     promoter        4258..4757
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     regulatory      4769..4834
                     /regulatory_class="promoter"
                     /note="EM7"
     promoter        4769..4816
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4835..5638
                     /codon_start=1
                     /product="neomycin resistance protein"
                     /label=neomycin resistance protein
                     /protein_id="AIA24531.1"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5676..5900
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(5957..5990)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     3UTR           6010..6511
                     /label=D114
                     /note="D114"
     promoter        complement(6533..6551)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6558..6574)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(6715..444)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"