LOCUS       sequence-69795-e        3371 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 3371)
  TITLE     Series-14: N-terminal Tag E.coli T7 Expression Vectors with Biobrick
            POLYPROMOTER Restriction Sites
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3371)
  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            "Unpublished"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 Sumo TEV expression vector with BioBr...
FEATURES             Location/Qualifiers
     source          1..3371
                     /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             310..603
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             610..630
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     terminator      672..718
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1885..1924
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1955..2543
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2444..2463
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2697..2714
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2729..2868
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2794..2813)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2954..2976
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2970..3161)
                     /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 atggctagca tgtcggactc agaagtcaat caagaagcta agccagaggt caagccagaa
      361 gtcaagcctg agactcacat caatttaaag gtgtccgatg gatcttcaga gatcttcttc
      421 aagatcaaaa agaccactcc tttaagaagg ctgatggaag cgttcgctaa aagacagggt
      481 aaggaaatgg actccttaag attcttgtac gacggtatta gaattcaagc tgatcagacc
      541 cctgaagatt tggacatgga ggataacgat attattgagg ctcacagaga acagattggt
      601 gggatcgagg aaaacctgta cttccaatcc aatattggaa gtggataact ggctgctgcc
      661 accgctgagc atagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
      721 gaaaggagga actatatcaa gtcccgaagt agctgtcaga ctgctggcga ccgccgcaac
      781 tagtggcgtc atgcgtgccg tatggtcttt cgcctgtaga cgccctacaa gccgtactaa
      841 agtggaaaga cccgcaaaag gtatcgcgat cgccctgcag ggaactatat ccgactagta
      901 gcccgcctaa tgagcgggct tttttttaat tcccctattt gtttattttt ctaaatacat
      961 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcgaaa aaggaagagt
     1021 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     1081 gtttttgctc acccagaaac gctcgtgaaa gtaaaagacg cagaggacca attgggggca
     1141 cgagtgggat acatagaact ggacttgaat agcggtaaaa tccttgagag ttttcgccct
     1201 gaagagcgtt ttccaatgat gagcactttc aaagttctgc tatgtggagc agtattatcc
     1261 cgtgtagatg cggggcaaga gcaactcgga cgacgaatac actattcgca gaatgacttg
     1321 gttgaatact ccccagtgac agaaaagcac cttacggacg gaatgacggt aagagaatta
     1381 tgtagtgccg ccataacgat gagtgataac actgcggcga acttacttct gacaaccatc
     1441 ggtggaccga aggaattaac cgcttttttg cacaatatgg gagaccatgt aactcgcctt
     1501 gaccgttggg aaccagaact gaatgaagcc ataccaaacg acgagcgaga caccacaatg
     1561 cctgcggcaa tggcaacaac attacgcaaa ctattaactg gcgaactact tactctggct
     1621 tcacggcaac aattaataga ctggcttgaa gcggataaag ttgcaggacc actactgcgt
     1681 tcggcacttc ctgctggctg gtttattgct gataaatctg gggcaggaga gcgtggttca
     1741 cggggtatca ttgccgcact tggaccagat ggtaagcctt cccgtatcgt agttatctac
     1801 acgacgggta gtcaggcaac tatggacgaa cgaaatagac agattgctga aataggggct
     1861 tcactgatta agcattggta aaccgataca attaaaggct ccttttggag cctttttttt
     1921 tggacggacc ggtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     1981 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2041 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2101 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2161 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     2221 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     2281 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     2341 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     2401 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     2461 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     2521 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     2581 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     2641 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     2701 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     2761 gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     2821 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     2881 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     2941 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     3001 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc
     3061 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
     3121 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
     3181 ggatttctgt tcatggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
     3241 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
     3301 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag
     3361 atgtaggtgt t
//
