LOCUS       sequence-69791-e        4169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4169)
  TITLE     Series-14: N-terminal Tag E.coli T7 Expression Vectors with Biobrick
            POLYPROMOTER Restriction Sites
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4169)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pET His6 MBP TEV expression vector with BioBri...
FEATURES             Location/Qualifiers
     source          1..4169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      48..94
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      154..181
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        207..225
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             242..264
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             283..300
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             307..1404
                     /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="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1378..1401
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1408..1428
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     terminator      1470..1516
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2683..2722
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      2753..3341
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3242..3261
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3495..3512
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3527..3666
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3592..3611)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3752..3774
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3768..3959)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ggcaccatgc atcgatcccg cgaaatgcgg ccgcttaatt aaggcatcaa ataaaacgaa
       61 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg agtccttcca
      121 gtctgggcga aagcgcggtc gatcaaatta agcagaaggc catcctgacg gatggccttt
      181 ttgcgtttct atcgatcccg cgaaattaat acgactcact ataggcccct ctagaaataa
      241 ttttgtttaa ctttaagaag gagatatacc atgggttctt ctcaccatca ccatcaccat
      301 ggttctaaaa tcgaagaagg taaactggta atctggatta acggcgataa aggctataac
      361 ggtctcgctg aagtcggtaa gaaattcgag aaagataccg gaattaaagt caccgttgag
      421 catccggata aactggaaga gaaattccca caggttgcgg caactggcga tggccctgac
      481 attatcttct gggcacacga ccgctttggt ggctacgctc aatctggcct gttggctgaa
      541 atcaccccgg acaaagcgtt ccaggacaag ctgtatccgt ttacctggga tgccgtacgt
      601 tacaacggca agctgattgc ttacccgatc gctgttgaag cgttatcgct gatttataac
      661 aaagatctgc tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct ggataaagaa
      721 ctgaaagcga aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg
      781 ccgctgattg ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt
      841 aaagacgtgg gcgtggataa cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg
      901 attaaaaaca aacacatgaa tgcagacacc gattactcca tcgcagaagc tgcctttaat
      961 aaaggcgaaa cagcgatgac catcaacggc ccgtgggcat ggtccaacat cgacaccagc
     1021 aaagtgaatt atggtgtaac ggtactgccg accttcaagg gtcaaccatc caaaccgttc
     1081 gttggcgtgc tgagcgcagg tattaacgcc gccagtccga acaaagagct ggcaaaagag
     1141 ttcctcgaaa actatctgct gactgatgaa ggtctggaag cggttaataa agacaaaccg
     1201 ctgggtgccg tagcgctgaa gtcttacgag gaagagttgg cgaaagatcc acgtattgcc
     1261 gccactatgg aaaacgccca gaaaggtgaa atcatgccga acatcccgca gatgtccgct
     1321 ttctggtatg ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat
     1381 gaagccctga aagacgcgca gactaatgaa aacctgtact tccaatccaa tattggaagt
     1441 ggataactgg ctgctgccac cgctgagcat agcataaccc cttggggcct ctaaacgggt
     1501 cttgaggggt tttttgctga aaggaggaac tatatcaagt cccgaagtag ctgtcagact
     1561 gctggcgacc gccgcaacta gtggcgtcat gcgtgccgta tggtctttcg cctgtagacg
     1621 ccctacaagc cgtactaaag tggaaagacc cgcaaaaggt atcgcgatcg ccctgcaggg
     1681 aactatatcc gactagtagc ccgcctaatg agcgggcttt tttttaattc ccctatttgt
     1741 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1801 cttcgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1861 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tcgtgaaagt aaaagacgca
     1921 gaggaccaat tgggggcacg agtgggatac atagaactgg acttgaatag cggtaaaatc
     1981 cttgagagtt ttcgccctga agagcgtttt ccaatgatga gcactttcaa agttctgcta
     2041 tgtggagcag tattatcccg tgtagatgcg gggcaagagc aactcggacg acgaatacac
     2101 tattcgcaga atgacttggt tgaatactcc ccagtgacag aaaagcacct tacggacgga
     2161 atgacggtaa gagaattatg tagtgccgcc ataacgatga gtgataacac tgcggcgaac
     2221 ttacttctga caaccatcgg tggaccgaag gaattaaccg cttttttgca caatatggga
     2281 gaccatgtaa ctcgccttga ccgttgggaa ccagaactga atgaagccat accaaacgac
     2341 gagcgagaca ccacaatgcc tgcggcaatg gcaacaacat tacgcaaact attaactggc
     2401 gaactactta ctctggcttc acggcaacaa ttaatagact ggcttgaagc ggataaagtt
     2461 gcaggaccac tactgcgttc ggcacttcct gctggctggt ttattgctga taaatctggg
     2521 gcaggagagc gtggttcacg gggtatcatt gccgcacttg gaccagatgg taagccttcc
     2581 cgtatcgtag ttatctacac gacgggtagt caggcaacta tggacgaacg aaatagacag
     2641 attgctgaaa taggggcttc actgattaag cattggtaaa ccgatacaat taaaggctcc
     2701 ttttggagcc tttttttttg gacggaccgg tagaaaagat caaaggatct tcttgagatc
     2761 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2821 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2881 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2941 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3001 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3061 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3121 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3181 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3241 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3301 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3361 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     3421 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     3481 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     3541 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg
     3601 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     3661 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     3721 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     3781 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     3841 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     3901 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     3961 gatgcctccg tgtaaggggg atttctgttc atggggtaat gataccgatg aaacgagaga
     4021 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     4081 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     4141 agcgcttcgt taatacagat gtaggtgtt
//
