LOCUS Exported 7341 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pSCB-YFP, complete sequence.
ACCESSION MH144608
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7341)
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 7341)
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 7341)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..7341
/organism="Cloning vector pSCB-YFP"
/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:2301261"
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..6256)
/codon_start=1
/product="aminoglycoside resistance protein"
/label=aminoglycoside resistance protein
/protein_id="AXM43032.1"
/translation="MVTAQWRFSWLLVMTCFFGVQSMPRASKQQARYAVGRCLMLWSSN
DVTQQGSRPKTKLNIMGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKP
HSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPA
KRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDL
FEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQ
PVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
CDS complement(5285..6076)
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
misc_feature 6275..20
/label=pSCB-YFP synthesis fragment
/note="pSCB-YFP synthesis fragment"
promoter 6392..6421
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 6429..6445
/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 6463..6474
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 6481..7200
/codon_start=1
/gene="EYFP"
/product="EYFP"
/label=EYFP
/protein_id="AXM43033.1"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
gene 6481..7200
/gene="EYFP"
/label=EYFP
terminator 7220..7291
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 7307..7334
/label=T7Te terminator
/note="phage T7 early transcription terminator"
LOCUS Exported 7341 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pSCB-YFP, complete sequence.
ACCESSION MH144608
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7341)
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 7341)
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 7341)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..7341
/organism="Cloning vector pSCB-YFP"
/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:2301261"
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..6256)
/codon_start=1
/product="aminoglycoside resistance protein"
/label=aminoglycoside resistance protein
/protein_id="AXM43032.1"
/translation="MVTAQWRFSWLLVMTCFFGVQSMPRASKQQARYAVGRCLMLWSSN
DVTQQGSRPKTKLNIMGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKP
HSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPA
KRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDL
FEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQ
PVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
CDS complement(5285..6076)
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
misc_feature 6275..20
/label=pSCB-YFP synthesis fragment
/note="pSCB-YFP synthesis fragment"
promoter 6392..6421
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 6429..6445
/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 6463..6474
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 6481..7200
/codon_start=1
/gene="EYFP"
/product="EYFP"
/label=EYFP
/protein_id="AXM43033.1"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
gene 6481..7200
/gene="EYFP"
/label=EYFP
terminator 7220..7291
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 7307..7334
/label=T7Te terminator
/note="phage T7 early transcription terminator"