LOCUS       40924_38423        5188 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pSABAD92A, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5188)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein 
            Sci."; date: "2010"; volume: "19"; issue: "8"; pages: "1534-1543"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (23-MAR-2010) Aalborg University, Sohngaardsholmsvej 49, Aalborg 
            9000, Denmark"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             572..589
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      798..884
                     /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
                     /label=AmpR promoter
     CDS             1113..1970
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2015..2470
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      2581..3169
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(3355..3495)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3947..4822)
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4849..5133
                     /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
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 tatgatcaaa atagccacac gtaaatattt aggcaaacaa aatgtctatg acattggagt
       61 tgagcgcgac cataattttg cactcaaaaa tggcttcata gcttctaatt gtttcaatgg
      121 taccggaagc gacggcgagc cactggtcgg cggcgacacg gacgaccagc tccagggcgg
      181 ctccggcgcc gatcgcctgg acggcggggc cggcgacgac atcctcgacg gcggcgccgg
      241 gcgcgaccgg ctgagcggcg gcgcgggcgc cgacaccttc gtgttctccg cccgcgagga
      301 cagctaccgt accgacacgg cggtgttcaa cgacctgatc ctcgacttcg aggccagcga
      361 ggatcgcatc gacctgtccg cgctgggctt ttccggcctg ggcgacggct atggcggcac
      421 cctgctcctg aagaccaacg ccgagggcac gcgcacctac ctgaaaagct tcgaggcgga
      481 tgccgaggga cggcgcttcg aggtcgccct ggacggcgac cacacgggcg atctttccgc
      541 cgccaatgtg gtcttcgccg cgaccgaatt ccatcatcat catcatcatt aagcttggct
      601 gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg
      661 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc
      721 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtccccc atgcgagagt
      781 agggaactgc caggcatcaa ataaaacgaa aggctcagtg caaagactgg gcctttcgtt
      841 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
      901 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
      961 ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc
     1021 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1081 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1141 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1201 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1261 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     1321 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     1381 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     1441 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     1501 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     1561 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     1621 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     1681 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     1741 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     1801 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     1861 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     1921 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     1981 accaagttta ctcatatata ctttagattg atttacgcgc cctgtagcgg cgcattaagc
     2041 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2101 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2161 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2221 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2281 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2341 ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2401 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     2461 tttacaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2521 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2581 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2641 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2701 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     2761 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2821 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2881 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2941 ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg
     3001 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3061 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3121 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3181 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3241 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3301 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     3361 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cggatatggt gcactctcag
     3421 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     3481 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3541 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3601 aggttttcac cgtcatcacc gaaacgcgcg aggcagcaag gagatggcgc ccaacagtcc
     3661 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     3721 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     3781 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atctaattct catgtttgac
     3841 agcttatcat cgatgcataa tgtgcctgtc aaatggacga agcagggatt ctgcaaaccc
     3901 tatgctactc cgtcaagccg tcaattgtct gattcgttac caattatgac aacttgacgg
     3961 ctacatcatt cactttttct tcacaaccgg cacggaactc gctcgggctg gccccggtgc
     4021 attttttaaa tacccgcgag aaatagagtt gatcgtcaaa accaacattg cgaccgacgg
     4081 tggcgatagg catccgggtg gtgctcaaaa gcagcttcgc ctggctgata cgttggtcct
     4141 cgcgccagct taagacgcta atccctaact gctggcggaa aagatgtgac agacgcgacg
     4201 gcgacaagca aacatgctgt gcgacgctgg cgatatcaaa attgctgtct gccaggtgat
     4261 cgctgatgta ctgacaagcc tcgcgtaccc gattatccat cggtggatgg agcgactcgt
     4321 taatcgcttc catgcgccgc agtaacaatt gctcaagcag atttatcgcc agcagctccg
     4381 aatagcgccc ttccccttgc ccggcgttaa tgatttgccc aaacaggtcg ctgaaatgcg
     4441 gctggtgcgc ttcatccggg cgaaagaacc ccgtattggc aaatattgac ggccagttaa
     4501 gccattcatg ccagtaggcg cgcggacgaa agtaaaccca ctggtgatac cattcgcgag
     4561 cctccggatg acgaccgtag tgatgaatct ctcctggcgg gaacagcaaa atatcacccg
     4621 gtcggcaaac aaattctcgt ccctgatttt tcaccacccc ctgaccgcga atggtgagat
     4681 tgagaatata acctttcatt cccagcggtc ggtcgataaa aaaatcgaga taaccgttgg
     4741 cctcaatcgg cgttaaaccc gccaccagat gggcattaaa cgagtatccc ggcagcaggg
     4801 gatcattttg cgcttcagcc atacttttca tactcccgcc attcagagaa gaaaccaatt
     4861 gtccatattg catcagacat tgccgtcact gcgtctttta ctggctcttc tcgctaacca
     4921 aaccggtaac cccgcttatt aaaagcattc tgtaacaaag cgggaccaaa gccatgacaa
     4981 aaacgcgtaa caaaagtgtc tataatcacg gcagaaaagt ccacattgat tatttgcacg
     5041 gcgtcacact ttgctatgcc atagcatttt tatccataag attagcggat cctacctgac
     5101 gctttttatc gcaactctct actgtttctc catacccgtt tttttgggct agaaataatt
     5161 ttgtttaact ttaagaagga gatataca
//
