LOCUS       Exported                3189 bp ds-DNA     circular SYN 29-NOV-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3189)
  AUTHORS   AG Stein
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Dec 2, 2019 from SnapGene Viewer 5.0.6
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3189
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     primer_bind     1..22
                     /label=control fw on bb
     primer_bind     complement(29..50)
                     /label=amp bb gblock2 rev
     primer_bind     complement(171..194)
                     /label=pFD amplify rv
     primer_bind     complement(178..194)
                     /label=oligo pFD mut 2
                     /note="auch zum amplifizieren des pFD gedacht"
     CDS             194..1292
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="ENRRR*TGNLD*RR*RL*RSG*SR*EIRERYRN*SHR*ASG*TGR
                     EIPTGCGNWRWP*HYLLGTRPLWWLRSIWPVG*NHPGQSVPGQAVSVYLGCRTLQRQAD
                     CLPDRC*SVIADL*QRSAAEPAKNLGRDPGAG*RTESER*ERADVQPARTVLHLAADCC
                     *RGLCVQV*KRQVRH*RRGRG*RWRESGSDLPG*PD*KQTHECRHRLLHRRSCL**RRN
                     SDDHQRPVGMVQHRHQQSELWCNGTADLQGSTIQTVRWRAERRY*RRQSEQRAGKRVPR
                     KLSAD**RSGSG**RQTAGCRSAEVLRGRVGERSAYCRHYGKRPER*NHAEHPADVRFL
                     VCRAYCGDQRRQRSSDCR*SPERRAD"
     primer_bind     194..216
                     /label=EcoRI for MBP
     primer_bind     complement(234..269)
                     /label=mbp bsa1 mut rv
     primer_bind     240..275
                     /label=mbp bsa1 mut fw
     primer_bind     complement(1281..1304)
                     /label=end MBP-link rv
     CDS             1305..1325
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1332..1355
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     misc_feature    1365..1388
                     /label=Autoinhibitory domain TVMV
     primer_bind     1389..1411
                     /note="color: black; sequence: GGCATTGCaTAAagccccgacaG"
     misc_feature    1398..1400
                     /label=final stop codon
     primer_bind     1435..1457
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1495..1513)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     CDS             complement(1646..2461)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2511..2538
                     /label=front of ori fw
     rep_origin      2589..3177
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3078..3097
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
       61 ttatcccctg attctgtgga taaccgtatt accgccaatt gagtgagctg ataccgctcg
      121 ccgcagccga acgaccgagc gcagcgagtc acccgcgaaa tacaagaaga caattttccc
      181 gcaccagcag tgggaaaatc gaagaaggta aactggtaat ctggattaac ggcgataaag
      241 gctataacgg tctggctgaa gtcggtaaga aattcgagaa agataccgga attaaagtca
      301 ccgttgagca tccggataaa ctggaagaga aattcccaca ggttgcggca actggcgatg
      361 gccctgacat tatcttctgg gcacacgacc gctttggtgg ctacgctcaa tctggcctgt
      421 tggctgaaat caccccggac aaagcgttcc aggacaagct gtatccgttt acctgggatg
      481 ccgtacgtta caacggcaag ctgattgctt acccgatcgc tgttgaagcg ttatcgctga
      541 tttataacaa agatctgctg ccgaacccgc caaaaacctg ggaagagatc ccggcgctgg
      601 ataaagaact gaaagcgaaa ggtaagagcg cgctgatgtt caacctgcaa gaaccgtact
      661 tcacctggcc gctgattgct gctgacgggg gttatgcgtt caagtatgaa aacggcaagt
      721 acgacattaa agacgtgggc gtggataacg ctggcgcgaa agcgggtctg accttcctgg
      781 ttgacctgat taaaaacaaa cacatgaatg cagacaccga ttactccatc gcagaagctg
      841 cctttaataa aggcgaaaca gcgatgacca tcaacggccc gtgggcatgg tccaacatcg
      901 acaccagcaa agtgaattat ggtgtaacgg tactgccgac cttcaagggt caaccatcca
      961 aaccgttcgt tggcgtgctg agcgcaggta ttaacgccgc cagtccgaac aaagagctgg
     1021 caaaagagtt cctcgaaaac tatctgctga ctgatgaagg tctggaagcg gttaataaag
     1081 acaaaccgct gggtgccgta gcgctgaagt cttacgagga agagttggtg aaagatccgc
     1141 gtattgccgc cactatggaa aacgcccaga aaggtgaaat catgccgaac atcccgcaga
     1201 tgtccgcttt ctggtatgcc gtgcgtactg cggtgatcaa cgccgccagc ggtcgtcaga
     1261 ctgtcgatga agccctgaaa gacgcgcaga ctaattcgag ctcggagaac ctgtatttcc
     1321 agtccagtgg ctggagccac ccgcagttcg aaaagtcagg aggtcgcgaa tatgtgcgct
     1381 ttgcgccggg cattgcataa agccccgaca gtaaaaagag accctgacta gtctccggga
     1441 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgacacga aagggcctcg
     1501 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     1561 gccttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     1621 tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca
     1681 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     1741 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     1801 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     1861 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt
     1921 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     1981 ccaaaccgtt attcattcgt gattgcgcct gagccagacg aaatacgcgg tcgctgttaa
     2041 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacaccgcc agcgcatcaa
     2101 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttccctggga
     2161 tcgccgtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     2221 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     2281 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
     2341 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     2401 catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca
     2461 taacacccct tgtattactg tttatgtaag cagacagttt tattgtgaat tctcatgacc
     2521 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2581 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2641 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2701 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     2761 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2821 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2881 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2941 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3001 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3061 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3121 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3181 gccagcaac
//
