LOCUS       Exported                8921 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFL8, complete sequence.
ACCESSION   KR061299
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8921)
  AUTHORS   Reeves AR, Weber G, Cernota WH, Weber JM.
  TITLE     Analysis of an 8.1-kb DNA fragment contiguous with the erythromycin 
            gene cluster of Saccharopolyspora erythraea in the eryCI-flanking 
            region
  JOURNAL   Antimicrob. Agents Chemother. 46 (12), 3892-3899 (2002)
  PUBMED    12435693
REFERENCE   2  (bases 1 to 8921)
  AUTHORS   Weber JM, Zieler H.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-APR-2015) Research and Development, Fermalogic, Inc., 
            4222 N. Ravenswood Ave., Suite 208, Chicago, IL 60613, USA
REFERENCE   3  (bases 1 to 8921)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8921
                     /organism="Cloning vector pFL8"
                     /mol_type="other DNA"
                     /db_xref="taxon:1718782"
     CDS             3426..4235
                     /codon_start=1
                     /gene="tsr"
                     /product="Tsr"
                     /label=tsr
                     /note="23S rRNA A1067 methyltransferase from Streptomyces 
                     azureus; thiostrepton resistance protein"
                     /protein_id="ALE71928.1"
                     /translation="MTELDTIANPSDPAVQRIIDVTKPSRSNIKTTLIEDVEPLMHSIA
                     AGVEFIEVYGSDSSPFPSELLDLCGRQNIPVRLIDSSIVNQLFKGERKAKTFGIARVPR
                     PARFGDIASRRGDVVVLDGVKIVGNIGAIVRTSLALGASGIILVDSDITSIADRRLQRA
                     SRGYVFSLPVVLSGREEAIAFIRDSGMQLMTLKADGDISVKELGDNPDRLALLFGSEKG
                     GPSDLFEEASSASVSIPMMSQTESLNVSVSLGIALHERIDRNLAANR"
     gene            3426..4235
                     /gene="tsr"
                     /label=tsr
     CDS             complement(4717..5511)
                     /codon_start=1
                     /gene="aph"
                     /product="Aph"
                     /label=aph
                     /note="aminoglycoside 3-phosphotransferase (APH) 
                     kanamycin/neomycin resistance protein"
                     /protein_id="ALE71929.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(4717..5511)
                     /gene="aph"
                     /label=aph
     rep_origin      complement(5774..6229)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(6206..6520)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSSKLTLTKGNKSLHACRSTLEDPRVPSSNSPYSESYYNS
                     LAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWTRPVAAH
                     "
     primer_bind     6370..6386
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6393..6411
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    6414..6470
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        complement(6477..6495)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6516..6532)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6540..6556
                     /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(6564..6594)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6609..6630
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6918..7506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7677..8537)
                     /codon_start=1
                     /gene="ampR"
                     /product="AmpR"
                     /label=ampR
                     /note="ampicillin resistance protein"
                     /protein_id="ALE71930.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(7677..8537)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(8538..8642)
                     /gene="bla"
                     /label=AmpR promoter