LOCUS       Exported                3932 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pBS-ldhGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3932)
  AUTHORS   Lizier M, Sarra PG, Cauda R, Lucchini F
  TITLE     Comparison of expression vectors in Lactobacillus reuteri strains.
  JOURNAL   FEMS Microbiol Lett. 2010 Jul 1. 308(1):8-15.
  PUBMED    20455948
REFERENCE   2  (bases 1 to 3932)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "FEMS 
            Microbiol Lett. 2010 Jul 1. 308(1):8-15."
FEATURES             Location/Qualifiers
     source          1..3932
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             6..722
                     /codon_start=1
                     /gene="mgfp5"
                     /product="GFP with folding enhancement mutations"
                     /label=mgfp5
                     /note="suitable for expression in plants due to removal of 
                     a cryptic intron"
                     /translation="MSKGEELFTGVVPILVELDGDVNGYKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKTRHNIEDGGVQLADHYQQNTPIGDDPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(36..64)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     659..679
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(724..740)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(724..740)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(772..791)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(773..791)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(801..818)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(801..817)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(810..832)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      959..1414
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1046..1065)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1256..1277
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1440..1544
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1545..2405
                     /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"
     primer_bind     complement(1763..1782)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2576..3164
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3065..3084
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3318..3335
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3452..3473
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3488..3518
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3526..3542
                     /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)."
     primer_bind     3531..3553
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3550..3566
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3550..3566
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3585..3605
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3587..3605
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     3634..3650
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3635..3651
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"