LOCUS       Exported                9120 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSh2-GFP, complete sequence.
ACCESSION   MF374793
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9120)
  AUTHORS   Lamont KC, Mudge SR, Liu G, Godwin ID.
  TITLE     Expression patterns of the native Shrunken-2 promoter in Sorghum 
            bicolor visualised through use of the GFP reporter gene
  JOURNAL   Plant Cell Rep. (2017) In press
  PUBMED    28721521
REFERENCE   2  (bases 1 to 9120)
  AUTHORS   Lamont KC, Mudge SR, Godwin ID.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-JUN-2017) School of Agriculture and Food Sciences, The
            University of Queensland, Mansfield Place, Brisbane, Queensland 
            4072, Australia
REFERENCE   3  (bases 1 to 9120)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9120
                     /organism="Cloning vector pSh2-GFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:2022454"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      413..2737
                     /regulatory_class="promoter"
     regulatory      2741..2750
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2747..3466
                     /codon_start=1
                     /gene="gfp"
                     /product="green fluorescent protein"
                     /label=gfp
                     /protein_id="ASN77188.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     gene            2747..3466
                     /gene="gfp"
                     /label=gfp
     regulatory      3472..4189
                     /regulatory_class="terminator"
                     /gene="octopine synthase"
     gene            3472..4189
                     /gene="octopine synthase"
                     /label=octopine synthase
     terminator      3475..4181
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     regulatory      4196..5604
                     /regulatory_class="terminator"
                     /gene="ScR1MYB1"
     gene            4196..5604
                     /gene="ScR1MYB1"
                     /label=ScR1MYB1
     regulatory      complement(5611..6845)
                     /regulatory_class="terminator"
                     /gene="rice actin"
     gene            complement(5611..6845)
                     /gene="rice actin"
                     /label=rice actin
     primer_bind     complement(6899..6915)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6923..6939
                     /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(6947..6977)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6992..7013
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7301..7889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8060..8920)
                     /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(8921..9025)
                     /gene="bla"
                     /label=AmpR promoter