LOCUS       Exported                6257 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHyVec7, complete sequence.
ACCESSION   EF539830
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6257)
  AUTHORS   Dana CE, Glauber KM, Chan TA, Bridge DM, Steele RE.
  TITLE     Incorporation of a horizontally transferred gene into an operon 
            during cnidarian evolution
  JOURNAL   PLoS ONE 7 (2), E31643 (2012)
  PUBMED    22328943
REFERENCE   2  (bases 1 to 6257)
  AUTHORS   Steele RE, Bridge DM, Chan TA, Dana K.
  TITLE     Vectors for expression of cloned genes in Hydra
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6257)
  AUTHORS   Steele RE, Bridge DM, Chan TA, Dana K.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-APR-2007) Department of Biological Chemistry, 
            University of California, Irvine, D240 Medical Sciences I, Irvine, 
            CA 92697-1700, USA
REFERENCE   4  (bases 1 to 6257)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6257
                     /organism="Cloning vector pHyVec7"
                     /mol_type="other DNA"
                     /db_xref="taxon:438597"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      654..1557
                     /regulatory_class="promoter"
                     /note="Hydra vulgaris actin"
     CDS             1666..2427
                     /codon_start=1
                     /gene="GFP"
                     /product="green fluorescent protein"
                     /label=GFP
                     /protein_id="ABP96788.1"
                     /translation="MADDEVAALAAGSMSKGEELFTGVVPILVELDGDVNGHKFSVSGE
                     GEGDATYGKLTLKFICTTGKLPVPWPTLVTTFCYGVQCFSRYPDHMKRHDFFKSAMPEG
                     YVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNV
                     YIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSK
                     DPNEKRDHMVLLEFVTAAGITHGMDELYKDP"
     gene            1666..2427
                     /gene="GFP"
                     /label=GFP
     CDS             1705..2418
                     /codon_start=1
                     /product="Aequoria victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFCYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    2428..2863
                     /note="intergenic region between Hydra replication factor C
                     large subunit gene and Hydra flp gene"
     misc_feature    2844..2845
                     /note="splice acceptor dinucleotide for trans-spliced 
                     leader"
     CDS             2864..3559
                     /codon_start=1
                     /gene="RFP"
                     /product="red fluorescent protein"
                     /label=RFP
                     /protein_id="ABP96789.1"
                     /translation="MADAAEMASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYE
                     GHNTVKLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMN
                     FEDGGVATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVL
                     KGETHKALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYER
                     TEGRHHLFL"
     gene            2864..3559
                     /gene="RFP"
                     /label=RFP
     misc_feature    3560..4055
                     /note="3 flanking sequence from Hydra vulgaris actin gene"
     regulatory      3691..3696
                     /regulatory_class="polyA_signal_sequence"
     misc_feature    4056..653
                     /label=derived from pBluescript II SK+
                     /note="derived from pBluescript II SK+"
     promoter        complement(4069..4087)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4108..4124)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4132..4148
                     /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(4156..4186)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4201..4222
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4510..5098)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5269..6129)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="ampicillin resistance protein"
                     /protein_id="ABP96790.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6130..6234)
                     /gene="bla"
                     /label=AmpR promoter