LOCUS       Exported               10843 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBS[NBL KO 1], complete sequence.
ACCESSION   MH187600
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10843)
  AUTHORS   Davis RJ, Belikoff EJ, Scholl EH, Li F, Scott MJ.
  TITLE     no blokes Is Essential for Male Viability and X Chromosome Gene 
            Expression in the Australian Sheep Blowfly
  JOURNAL   Curr. Biol. (2018) In press
  PUBMED    29887311
REFERENCE   2  (bases 1 to 10843)
  AUTHORS   Davis RJ, Belikoff EJ, Scholl EH, Li F, Scott MJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-APR-2018) Entomology and Plant Pathology, North 
            Carolina State University, 112 Derieux Place, 1546 Thomas Hall, 
            Raleigh, NC 27607, USA
REFERENCE   3  (bases 1 to 10843)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..10843
                     /organism="Cloning vector pBS[NBL KO 1]"
                     /mol_type="other DNA"
                     /db_xref="taxon:2231719"
     misc_feature    complement(1..1133)
                     /label=homology arm derived from Lucilia cuprina NBL
                     /note="homology arm derived from Lucilia cuprina NBL"
     promoter        complement(1164..1182)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1203..1219)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1227..1243
                     /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(1251..1281)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1296..1317
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1605..2193)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2364..3224)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3225..3329)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3355..3810)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     3952..3968
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3978..3996
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(4008..5073)
                     /label=homology arm derived from Lucilia cuprina NBL
                     /note="homology arm derived from Lucilia cuprina NBL"
     regulatory      5074..8495
                     /regulatory_class="promoter"
                     /note="derived from Lucilia cuprina h83"
     CDS             8496..9173
                     /codon_start=1
                     /product="DsRedexpress2"
                     /label=DsRedexpress2
                     /protein_id="AWV91982.1"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     misc_feature    9174..10599
                     /note="derived from Lucilia cuprina atub UTR and 
                     polyadenylation signal sequence"
     misc_recomb     10600..10843
                     /note="phic31; attB"
                     /note="/recombination_class=other"
     protein_bind    10663..10762
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"