LOCUS       Exported                5188 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pSABAD92A, complete sequence.
ACCESSION   HM070247
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5188)
  AUTHORS   Buchinger E, Aachmann FL, Aranko AS, Valla S, Skjak-Braek G, Iwai H,
            Wimmer R.
  TITLE     Use of protein trans-splicing to produce active and segmentally 
            (2)H, (15)N labeled mannuronan C5-epimerase AlgE4
  JOURNAL   Protein Sci. 19 (8), 1534-1543 (2010)
  PUBMED    20552686
REFERENCE   2  (bases 1 to 5188)
  AUTHORS   Buchinger E, Aachmann FL, Aranko S, Valla S, Sjaak-Braek G, Iwai H, 
            Wimmer R.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-MAR-2010) Aalborg University, Sohngaardsholmsvej 49, 
            Aalborg 9000, Denmark
REFERENCE   3  (bases 1 to 5188)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5188
                     /organism="Expression vector pSABAD92A"
                     /mol_type="other DNA"
                     /db_xref="taxon:762607"
     CDS             2..592
                     /codon_start=1
                     /product="IntC-R-H6"
                     /label=IntC-R-H6
                     /note="fusion protein of Nostoc punctiforme DnaE intein 
                     (IntC) C-terminal fragment and Azotobacter vinelandii 
                     mannuronan C5-epimerase AlgE4 R-module (R) plus His-tag"
                     /protein_id="ADG44967.1"
                     /translation="MIKIATRKYLGKQNVYDIGVERDHNFALKNGFIASNCFNGTGSDG
                     EPLVGGDTDDQLQGGSGADRLDGGAGDDILDGGAGRDRLSGGAGADTFVFSAREDSYRT
                     DTAVFNDLILDFEASEDRIDLSALGFSGLGDGYGGTLLLKTNAEGTRTYLKSFEADAEG
                     RRFEVALDGDHTGDLSAANVVFAATEFHHHHHH"
     CDS             572..589
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      798..884
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      976..1003
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1021..1112
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1113..1973
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2015..2470
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      2581..3169
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3355..3495
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3944..4822)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4849..5133
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     RBS             5160..5182
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"