LOCUS       sequence-344699-        3414 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 N-terminal tag module carrying 6xHIS-MBP-3C .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3414)
  TITLE     Banfield lab plasmids
REFERENCE   2  (bases 1 to 3414)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pICSL30015
FEATURES             Location/Qualifiers
     source          1..3414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(160..951)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(1173..1277)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1345..1363
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             1405..1422
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1432..2532
                     /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="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     2506..2529
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2542..2565
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2644..2661)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2815..3403)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2895..2914)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
       61 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      121 atcaatctaa agtatatatg agtaaacttg gtctgacagt tatttgccga ctaccttggt
      181 gatctcgcct tttacgtaat ggacaaattc ttccaactga tctgcgcgcg aggccaagcg
      241 atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc
      301 cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac
      361 tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc
      421 gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg
      481 aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc
      541 ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag
      601 aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg
      661 aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc
      721 aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag
      781 ccttacggtc accgtaacca gcaaatcaat atcgctgtgt ggcttcaggc cgccatccac
      841 tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc
      901 aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccctca tgatgtttaa
      961 ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctccata acatcaaaca
     1021 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtaccccaaa
     1081 aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg
     1141 gtcaaggttc tggaccagtt gcgtgagcgc atactcttcc tttttcaata ttattgaagc
     1201 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     1261 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     1321 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tccactgaag
     1381 agccactgtg gtctcaccat ggcacaccat caccaccatc acagcagcgg catgaaaatc
     1441 gaagaaggta aactggtaat ctggattaac ggcgataaag gctataacgg cctcgctgaa
     1501 gtcggtaaga aattcgagaa agataccgga attaaagtca ccgttgagca tccggataaa
     1561 ctggaagaga aattcccaca ggttgcggca actggcgatg gccctgacat tatcttctgg
     1621 gcacacgacc gctttggtgg ctacgctcaa tctggcctgt tggctgaaat caccccggac
     1681 aaagcgttcc aggacaagct gtatccgttt acctgggatg ccgtacgtta caacggcaag
     1741 ctgattgctt acccgatcgc tgttgaagcg ttatcgctga tttataacaa agatctgctg
     1801 ccgaacccgc caaaaacctg ggaagagatc ccggcgctgg ataaagaact gaaagcgaaa
     1861 ggtaagagcg cgctgatgtt caacctgcaa gaaccgtact tcacctggcc gctgattgct
     1921 gctgacgggg gttatgcgtt caagtatgaa aacggcaagt acgacattaa agacgtgggc
     1981 gtggataacg ctggcgcgaa agcgggtctg accttcctgg ttgacctgat taaaaacaaa
     2041 cacatgaatg cagacaccga ttactccatc gcagaagctg cctttaataa aggcgaaaca
     2101 gcgatgacca tcaacggccc gtgggcatgg tccaacatcg acaccagcaa agtgaattat
     2161 ggtgtaacgg tactgccgac cttcaagggt caaccatcca aaccgttcgt tggcgtgctg
     2221 agcgcaggta ttaacgccgc cagtccgaac aaagagctgg caaaagagtt cctcgaaaac
     2281 tatctgctga ctgatgaagg tctggaagcg gttaataaag acaaaccgct gggtgccgta
     2341 gcgctgaagt cttacgagga agagttggcg aaagatccac gtattgccgc cactatggaa
     2401 aacgcccaga aaggtgaaat catgccgaac atcccgcaga tgtccgcttt ctggtatgcc
     2461 gtgcgtactg cggtgatcaa cgccgccagc ggtcgtcaga ctgtcgatga agccctgaaa
     2521 gacgcgcaga ctagcagcgg tctggaagtt ctgtttcagg gcccaatgtg agaccacgag
     2581 tgattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2641 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2701 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2761 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2821 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2881 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2941 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3001 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3061 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3121 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3181 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3241 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3301 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3361 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc cttt
//
