LOCUS       Exported                6633 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSCBe, complete sequence.
ACCESSION   MH144609
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6633)
  AUTHORS   Jin H, Wang Y, Idoine A, Bhaya D.
  TITLE     Construction of a Shuttle Vector Using an Endogenous Plasmid From 
            the Cyanobacterium Synechocystis sp. PCC6803
  JOURNAL   Front Microbiol 9, 1662 (2018)
  PUBMED    30087668
REFERENCE   2  (bases 1 to 6633)
  AUTHORS   Haojie J, Devaki B.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-MAR-2018) Plant Biology Department, Carnegie 
            Institution for Science, 260 Panama Street, Stanford, CA 94305, US
REFERENCE   3  (bases 1 to 6633)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6633
                     /organism="Cloning vector pSCBe"
                     /mol_type="other DNA"
                     /country="USA"
                     /isolation_source="fresh water"
                     /label=derived from Synechocystis sp. PCC 6803
                     /note="derived from Synechocystis sp. PCC 6803"
                     /db_xref="taxon:2301262"
     gene            258..3173
                     /gene="ORFb"
                     /label=ORFb
     CDS             3798..3989
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     oriT            4090..4230
     misc_feature    4091..4231
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      4402..5017
     rep_origin      complement(4417..5005)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5285..6076)
                     /codon_start=1
                     /product="aminoglycoside resistance protein"
                     /label=aminoglycoside resistance protein
                     /protein_id="AXM43034.1"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    6267..6532
                     /label=MCS
                     /note="MCS"
     promoter        6298..6327
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    6335..6351
                     /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)."
     RBS             6369..6380
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             6390..6413
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6477..6494
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      6512..6583
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6599..6626
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"