LOCUS       Exported                5289 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCS2plus_mVenus-Cdt1, complete sequence.
ACCESSION   MF614950
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5289)
  AUTHORS   Ozpolat BD, Handberg-Thorsager M, Vervoort M, Balavoine G.
  TITLE     Cell lineage and cell cycling analyses of the 4d micromere using 
            live imaging in the marine annelid Platynereis dumerilii
  JOURNAL   Elife 6 (2017) In press
  PUBMED    29231816
REFERENCE   2  (bases 1 to 5289)
  AUTHORS   Ozpolat BD, Handberg-Thorsager M, Vervoort M, Balavoine G.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-AUG-2017) Marine Biological Laboratory, Eugene Bell 
            Center for Regenerative Biology and Tissue Engineering, 7 Mbl 
            Street, Mrc 212, Woods Hole, MA 02543, USA
REFERENCE   3  (bases 1 to 5289)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5289
                     /organism="Cloning vector pCS2plus_mVenus-Cdt1"
                     /mol_type="other DNA"
                     /PCR_primers="fwd_name: pcs2+f1, fwd_seq:
                     ctagaactatagtgtgttgtattacgt, rev_name: pcs2+r1, rev_seq: 
                     agaggccttgaattcgaatcg"
                     /db_xref="taxon:2060959"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             119..1300
                     /codon_start=1
                     /product="mVenus-Cdt1"
                     /label=mVenus-Cdt1
                     /protein_id="AUG68261.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKSGLRSRAMAQTSVTNFFASRKRGADIHASKRRKVQRLTQDVE
                     TENVSVSEQANAELQSSRPAGKGDLKVPELQKNEVGEDSSTATKPSTRKSAPRATKTRK
                     ASTSTRSSQRSKTKAAASAGQKDITQFLSSILESDDKTKDVLPEPEPTAAIDD"
     misc_feature    119..856
                     /label=Region: mVenus
                     /note="Region: mVenus"
     CDS             119..835
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    857..1297
                     /note="Region: Cdt1; derived from Platynereis dumerilii"
     polyA_signal    1338..1472
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(1585..1603)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1624..1640)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1648..1664
                     /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)."
     promoter        complement(1672..1702)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1717..1738
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2026..2614)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2785..3645)
                     /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"
     promoter        complement(3646..3750)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3776..4231)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4305..5289
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"