LOCUS       Exported                4151 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pEG, complete sequence.
ACCESSION   KT962975
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4151)
  AUTHORS   Zhou Y, Zhang L, Shah N, Palmer R Jr.
  TITLE     Cell-surface glycoside hydrolases of Streptococcus gordonii promote 
            growth in saliva
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4151)
  AUTHORS   Yang J, Cisar JO.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-OCT-2015) Laboratory of Cell and Developmental 
            Biology, National Institute of Health, 9000 Rockville Pike, 
            Bethesda, MD 20854, USA
REFERENCE   3  (bases 1 to 4151)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4151
                     /organism="Cloning vector pEG"
                     /mol_type="other DNA"
                     /db_xref="taxon:1775243"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      402..599
                     /regulatory_class="promoter"
                     /note="consititutive promoter located upstream of 
                     Streptococcus gordonii rpsJ gene"
     CDS             600..1316
                     /codon_start=1
                     /gene="GFP"
                     /product="green fluorecent protein"
                     /label=GFP
                     /note="codon optimized"
                     /protein_id="ALU57381.1"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            600..1316
                     /gene="GFP"
                     /label=GFP
     misc_feature    1320..1376
                     /label=multiple cloning site
                     /note="multiple cloning site"
     primer_bind     complement(1389..1405)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1413..1429
                     /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(1437..1467)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1482..1503
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      1776..2376
                     /label=ori of pUC19
                     /note="ori of pUC19"
     rep_origin      complement(1791..2379)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             2951..3688
                     /codon_start=1
                     /gene="ermAM"
                     /product="erythromycin resistant protein"
                     /label=ermAM
                     /protein_id="ALU57382.1"
                     /translation="MNKNIKYSQNFLTSEKVLNQIIKQLNLKETDTVYEIGTGKGHLTT
                     KLAKISKQVTSIELDSHLFNLSSEKLKLNTRVTLIHQDILQFQFPNKQRYKIVGNIPYH
                     LSTQIIKKVVFESHASDIYLIVEEGFYKRTLDIHRTLGLLLHTQVSIQQLLKLPAECFH
                     PKPKVNSVLIKLTRHTTDVPDKYWKLYTYFVSKWVNREYRQLFTKNQFHQAMKHAKVNN
                     LSTVTYEQVLSIFNSYLLFNGRK"
     gene            2951..3688
                     /gene="ermAM"
                     /label=ermAM
     promoter        complement(3952..4056)
                     /gene="bla"
                     /label=AmpR promoter