LOCUS       Exported                3813 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRL446, complete sequence.
ACCESSION   EU346690
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3813)
  AUTHORS   Elhai J, Wolk CP.
  TITLE     A versatile class of positive-selection vectors based on the 
            nonviability of palindrome-containing plasmids that allows cloning 
            into long polylinkers
  JOURNAL   Gene 68 (1), 119-138 (1988)
  PUBMED    2851487
REFERENCE   2  (bases 1 to 3813)
  AUTHORS   Zhang W, Du Y, Khudyakov I, Fan Q, Gao H, Ning D, Wolk CP, Xu X.
  TITLE     A gene cluster that regulates both heterocyst differentiation and 
            pattern formation in Anabaena sp. strain PCC 7120
  JOURNAL   Mol. Microbiol. 66 (6), 1429-1443 (2007)
  PUBMED    18045384
REFERENCE   3  (bases 1 to 3813)
  AUTHORS   Elhai J, Xu X, Wolk CP.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-DEC-2007) MSU-DOE Plant Research Laboratory and 
            Department of Plant Biology, Michigan State University, Wilson Road,
            E. Lansing, MI 48824, USA
REFERENCE   4  (bases 1 to 3813)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3813
                     /organism="Cloning vector pRL446"
                     /mol_type="other DNA"
                     /db_xref="taxon:491925"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             562..1377
                     /codon_start=1
                     /gene="npt"
                     /product="neomycin phosphotransferase"
                     /label=npt
                     /note="antibiotic-resistance protein"
                     /protein_id="ABY59252.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            562..1377
                     /gene="npt"
                     /label=npt
     primer_bind     complement(1592..1608)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1616..1632
                     /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(1640..1670)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1685..1706
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1994..2582)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2753..3613)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="antibiotic-resistance protein"
                     /protein_id="ABY59253.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(2753..3613)
                     /gene="bla"
                     /label=bla
     promoter        complement(3614..3718)
                     /gene="bla"
                     /label=AmpR promoter