LOCUS       Exported                9669 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector L274-12, complete sequence.
ACCESSION   JF419120
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9669)
  AUTHORS   Sommer F, Awazu S, Anton-Erxleben F, Jiang D, Klimovich AV, 
            Klimovich BV, Samoilovich MP, Satou Y, Kruss M, Gelhaus C, Kurn U, 
            Bosch TC, Khalturin K.
  TITLE     Blood System Formation in the Urochordate Ciona intestinalis 
            Requires the Variable Receptor vCRL1
  JOURNAL   Mol. Biol. Evol. 29 (10), 3081-3093 (2012)
  PUBMED    22513285
REFERENCE   2  (bases 1 to 9669)
  AUTHORS   Sommer F, Awazu S, Anton-Erxleben F, Jiang D, Klimovich AV, 
            Klimovich VB, Samoilovich MP, Satou Y, Kruess M, Gelhaus C, Kuern U,
            Bosch TCG., Khalturin K.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-FEB-2011) Zoological Institute, Christian-Albrechts 
            University Kiel, Am Botanischen Garten 9, Kiel, Schleswig-Holstein 
            24118, Germany
REFERENCE   3  (bases 1 to 9669)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9669
                     /organism="Expression vector L274-12"
                     /mol_type="other DNA"
                     /note="pMiLRneo::EF1a_Prom::EGFP::vCRL1_hp::EF1a_Term(L274 
                     -12)"
                     /db_xref="taxon:997209"
     CDS             2448..3161
                     /codon_start=1
                     /product="Aequoria victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFCYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     regulatory      4119..4128
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        complement(4159..4177)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4184..4200)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6826..6844)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6854..6870)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      7012..7467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        7493..7597
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7598..8458
                     /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      8629..9217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    9505..9526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9541..9571
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9579..9595
                     /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     9603..9619
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        9640..9658
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"