LOCUS       Exported                5374 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Diatom vector pDEST-N-HA, complete sequence.
ACCESSION   LT726868
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5374)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5374)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 5374)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5374
                     /organism="Diatom vector pDEST-N-HA"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 7881."
                     /db_xref="taxon:1944723"
     rep_origin      complement(6..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 ori"
     primer_bind     603..619
                     /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      626..642
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     regulatory      865..1100
                     /regulatory_class="promoter"
                     /note="FcpB promoter"
     misc_feature    1114..1140
                     /label=HA epitope
                     /note="HA epitope"
     CDS             1114..1140
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    1141..1167
                     /label=HA epitope
                     /note="HA epitope"
     CDS             1141..1167
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    1183..1209
                     /label=HA epitope
                     /note="HA epitope"
     CDS             1183..1209
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      complement(1225..1349)
                     /regulatory_class="attenuator"
                     /note="attR1"
     protein_bind    1225..1349
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      1357..1409
                     /regulatory_class="promoter"
                     /note="lacUV5 promoter"
     promoter        1374..1404
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1458..2117
                     /codon_start=1
                     /note="unnamed protein product; chloramphenicol resistance"
                     /protein_id="SJL86720.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2459..2764
                     /codon_start=1
                     /note="unnamed protein product; ccdB"
                     /protein_id="SJL86721.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     regulatory      2805..2929
                     /regulatory_class="attenuator"
                     /note="attR2"
     protein_bind    complement(2805..2929)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      3097..3170
                     /regulatory_class="terminator"
                     /note="FcpA terminator"
     promoter        complement(3189..3207)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      complement(3191..3207)
                     /regulatory_class="promoter"
                     /note="T3 promoter"
     primer_bind     complement(3228..3244)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(3233..3270)
                     /label=lacZ RBS
                     /note="lacZ RBS"
     protein_bind    3252..3268
                     /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)."
     regulatory      complement(3271..3354)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(3276..3306)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3321..3342
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3446..3848)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(3630..4218)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4389..5249)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL86722.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5250..5354)
                     /gene="bla"
                     /label=AmpR promoter