LOCUS       sequence-69793-e        4151 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 4151)
  TITLE     Series-14: N-terminal Tag E.coli T7 Expression Vectors with Biobrick
            POLYPROMOTER Restriction Sites
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4151)
  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            "Unpublished"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 MBP TEV expression vector with BrioBrick
            p...
FEATURES             Location/Qualifiers
     source          1..4151
                     /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             289..1386
                     /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     1360..1383
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1390..1410
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     terminator      1452..1498
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2665..2704
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      2735..3323
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3224..3243
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3477..3494
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3509..3648
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3574..3593)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3734..3756
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3750..3941)
                     /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 ctggttctaa aatcgaagaa
      301 ggtaaactgg taatctggat taacggcgat aaaggctata acggtctcgc tgaagtcggt
      361 aagaaattcg agaaagatac cggaattaaa gtcaccgttg agcatccgga taaactggaa
      421 gagaaattcc cacaggttgc ggcaactggc gatggccctg acattatctt ctgggcacac
      481 gaccgctttg gtggctacgc tcaatctggc ctgttggctg aaatcacccc ggacaaagcg
      541 ttccaggaca agctgtatcc gtttacctgg gatgccgtac gttacaacgg caagctgatt
      601 gcttacccga tcgctgttga agcgttatcg ctgatttata acaaagatct gctgccgaac
      661 ccgccaaaaa cctgggaaga gatcccggcg ctggataaag aactgaaagc gaaaggtaag
      721 agcgcgctga tgttcaacct gcaagaaccg tacttcacct ggccgctgat tgctgctgac
      781 gggggttatg cgttcaagta tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat
      841 aacgctggcg cgaaagcggg tctgaccttc ctggttgacc tgattaaaaa caaacacatg
      901 aatgcagaca ccgattactc catcgcagaa gctgccttta ataaaggcga aacagcgatg
      961 accatcaacg gcccgtgggc atggtccaac atcgacacca gcaaagtgaa ttatggtgta
     1021 acggtactgc cgaccttcaa gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca
     1081 ggtattaacg ccgccagtcc gaacaaagag ctggcaaaag agttcctcga aaactatctg
     1141 ctgactgatg aaggtctgga agcggttaat aaagacaaac cgctgggtgc cgtagcgctg
     1201 aagtcttacg aggaagagtt ggcgaaagat ccacgtattg ccgccactat ggaaaacgcc
     1261 cagaaaggtg aaatcatgcc gaacatcccg cagatgtccg ctttctggta tgccgtgcgt
     1321 actgcggtga tcaacgccgc cagcggtcgt cagactgtcg atgaagccct gaaagacgcg
     1381 cagactaatg aaaacctgta cttccaatcc aatattggaa gtggataact ggctgctgcc
     1441 accgctgagc atagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     1501 gaaaggagga actatatcaa gtcccgaagt agctgtcaga ctgctggcga ccgccgcaac
     1561 tagtggcgtc atgcgtgccg tatggtcttt cgcctgtaga cgccctacaa gccgtactaa
     1621 agtggaaaga cccgcaaaag gtatcgcgat cgccctgcag ggaactatat ccgactagta
     1681 gcccgcctaa tgagcgggct tttttttaat tcccctattt gtttattttt ctaaatacat
     1741 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcgaaa aaggaagagt
     1801 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     1861 gtttttgctc acccagaaac gctcgtgaaa gtaaaagacg cagaggacca attgggggca
     1921 cgagtgggat acatagaact ggacttgaat agcggtaaaa tccttgagag ttttcgccct
     1981 gaagagcgtt ttccaatgat gagcactttc aaagttctgc tatgtggagc agtattatcc
     2041 cgtgtagatg cggggcaaga gcaactcgga cgacgaatac actattcgca gaatgacttg
     2101 gttgaatact ccccagtgac agaaaagcac cttacggacg gaatgacggt aagagaatta
     2161 tgtagtgccg ccataacgat gagtgataac actgcggcga acttacttct gacaaccatc
     2221 ggtggaccga aggaattaac cgcttttttg cacaatatgg gagaccatgt aactcgcctt
     2281 gaccgttggg aaccagaact gaatgaagcc ataccaaacg acgagcgaga caccacaatg
     2341 cctgcggcaa tggcaacaac attacgcaaa ctattaactg gcgaactact tactctggct
     2401 tcacggcaac aattaataga ctggcttgaa gcggataaag ttgcaggacc actactgcgt
     2461 tcggcacttc ctgctggctg gtttattgct gataaatctg gggcaggaga gcgtggttca
     2521 cggggtatca ttgccgcact tggaccagat ggtaagcctt cccgtatcgt agttatctac
     2581 acgacgggta gtcaggcaac tatggacgaa cgaaatagac agattgctga aataggggct
     2641 tcactgatta agcattggta aaccgataca attaaaggct ccttttggag cctttttttt
     2701 tggacggacc ggtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2761 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2821 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2881 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2941 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3001 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3061 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3121 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3181 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3241 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3301 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3361 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     3421 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3481 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     3541 gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3601 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     3661 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     3721 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     3781 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc
     3841 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
     3901 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
     3961 ggatttctgt tcatggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
     4021 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
     4081 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag
     4141 atgtaggtgt t
//
