LOCUS       Exported                4798 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Integrative expression vector pFPN, complete sequence.
ACCESSION   EF468631
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4798)
  AUTHORS   Chaurasia AK, Parasnis A, Apte SK.
  TITLE     An integrative expression vector for strain improvement and 
            environmental applications of the nitrogen fixing cyanobacterium, 
            Anabaena sp. strain PCC7120
  JOURNAL   J. Microbiol. Methods 73 (2), 133-141 (2008)
  PUBMED    18367274
REFERENCE   2  (bases 1 to 4798)
  AUTHORS   Chaurasia AK, Parasnis A, Apte SK.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-MAR-2007) Molecular Biology Division, Bhabha Atomic 
            Research Centre, Trombay, Mumbai, Maharashtra 400 085, India
REFERENCE   3  (bases 1 to 4798)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4798
                     /organism="Integrative expression vector pFPN"
                     /mol_type="other DNA"
                     /db_xref="taxon:433946"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     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"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    707..2564
                     /label=integrative expression cassette
                     /note="integrative expression cassette"
     misc_feature    721..1221
                     /note="F1 flanking region; derived from Anabaena sp. 
                     PCC7120"
     regulatory      1224..1388
                     /regulatory_class="promoter"
                     /note="light inducible PpsbA1 promoter from Anabaena sp. 
                     PCC7120"
     CDS             1458..2189
                     /codon_start=1
                     /gene="nptII"
                     /product="neomycin phosphotransferase"
                     /label=nptII
                     /note="derived from pDUCA7"
                     /protein_id="ABO72539.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRPPILKPITILLGSEDLP"
     gene            1458..2189
                     /gene="nptII"
                     /label=nptII
     misc_feature    2139..2570
                     /note="F2 flanking region; derived from Anabaena sp. 
                     PCC7120"
     promoter        complement(2607..2625)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2646..2662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2670..2686
                     /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(2694..2724)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2739..2760
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2866..3806
                     /note="ColE1 origin of replication; derived from 
                     pBluescript SK(+/-)"
     rep_origin      complement(3048..3636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3807..4667)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="provides ampicllin resistance; derived from 
                     pBluescript SK(+/-)"
                     /protein_id="ABO72540.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3807..4667)
                     /gene="bla"
                     /label=bla
     promoter        complement(4668..4772)
                     /gene="bla"
                     /label=AmpR promoter