LOCUS       Exported                6861 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLAL1, complete sequence.
ACCESSION   KP696472
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6861)
  AUTHORS   Lacerda LA, Ferreira H.
  TITLE     Xanthomonas citri Expression Vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6861)
  AUTHORS   Lacerda LA, Ferreira H.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-JAN-2015) Bioquimica e Microbiologia, UNESP, Av. 24A, 
            1515, Rio Claro, SP 13506900, Brazil
REFERENCE   3  (bases 1 to 6861)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6861
                     /organism="Expression vector pLAL1"
                     /mol_type="other DNA"
                     /db_xref="taxon:1744848"
     rep_origin      1023..1792
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1793..2455
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     primer_bind     3162..3178
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3188..3206
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3239..3255
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(3358..4236)
                     /regulatory_class="promoter"
                     /gene="araC"
     gene            complement(3358..4236)
                     /gene="araC"
                     /label=araC
     CDS             complement(3358..4236)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4263..4547
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     CDS             4582..5370
                     /codon_start=1
                     /gene="acp-TAP"
                     /product="acyl carrier protein-TAP"
                     /label=acp-TAP
                     /protein_id="ALL35039.1"
                     /translation="MSTIEERVKKIIGEQLGVKQEEVTNNASFVEDLGADSLDTVELVM
                     ALEEEFDTEIPDEEAEKITTVQAAIDYINGHQASMEKRRWKKNFIAVSAANRFKKISSS
                     GALDYDIPTTASENLYFQGELKTAALAQHDEAVDNKFNKEQQNAFYEILHLPNLNEEQR
                     NAFIQSLKDDPSQSANLLAEAKKLNDAQAPKVDNKFNKEQQNAFYEILHLPNLNEEQRN
                     AFIQSLKDDPSQSANLLAEAKKLNGAQAPKVDANSAGKST"
     gene            4582..5370
                     /gene="acp-TAP"
                     /label=acp-TAP
     CDS             4582..4815
                     /codon_start=1
                     /gene="acpP"
                     /product="acyl carrier protein (ACP) from E. coli"
                     /label=ACP
                     /translation="MSTIEERVKKIIGEQLGVKQEEVTNNASFVEDLGADSLDTVELVM
                     ALEEEFDTEIPDEEAEKITTVQAAIDYINGHQA"
     CDS             4825..4902
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             4930..4950
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4990..5163
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             5167..5337
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNGAQAPK"
     primer_bind     complement(5378..5394)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5424..5442)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5463..5479)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5487..5503
                     /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(5511..5541)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5556..5577
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5851..5879
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             6068..6601
                     /codon_start=1
                     /product="gentamicin-(3)-N-acetyl-transferase"
                     /label=gentamicin-
                     /note="confers gentamicin resistance"
                     /protein_id="ALL35038.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"