LOCUS       sequence-69790-e        3062 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 3062)
  TITLE     Series-14: N-terminal Tag E.coli T7 Expression Vectors with Biobrick
            POLYPROMOTER Restriction Sites
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3062)
  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            "Unpublished"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 TEV expression vector with BioBrick
            p...
FEATURES             Location/Qualifiers
     source          1..3062
                     /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             301..321
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     terminator      363..409
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1576..1615
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1646..2234
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2135..2154
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2388..2405
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2420..2559
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2485..2504)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2645..2667
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2661..2852)
                     /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 gaaaacctgt acttccaatc caatattgga agtggataac tggctgctgc caccgctgag
      361 catagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
      421 aactatatca agtcccgaag tagctgtcag actgctggcg accgccgcaa ctagtggcgt
      481 catgcgtgcc gtatggtctt tcgcctgtag acgccctaca agccgtacta aagtggaaag
      541 acccgcaaaa ggtatcgcga tcgccctgca gggaactata tccgactagt agcccgccta
      601 atgagcgggc ttttttttaa ttcccctatt tgtttatttt tctaaataca ttcaaatatg
      661 tatccgctca tgagacaata accctgataa atgcttcgaa aaaggaagag tatgagtatt
      721 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
      781 cacccagaaa cgctcgtgaa agtaaaagac gcagaggacc aattgggggc acgagtggga
      841 tacatagaac tggacttgaa tagcggtaaa atccttgaga gttttcgccc tgaagagcgt
      901 tttccaatga tgagcacttt caaagttctg ctatgtggag cagtattatc ccgtgtagat
      961 gcggggcaag agcaactcgg acgacgaata cactattcgc agaatgactt ggttgaatac
     1021 tccccagtga cagaaaagca ccttacggac ggaatgacgg taagagaatt atgtagtgcc
     1081 gccataacga tgagtgataa cactgcggcg aacttacttc tgacaaccat cggtggaccg
     1141 aaggaattaa ccgctttttt gcacaatatg ggagaccatg taactcgcct tgaccgttgg
     1201 gaaccagaac tgaatgaagc cataccaaac gacgagcgag acaccacaat gcctgcggca
     1261 atggcaacaa cattacgcaa actattaact ggcgaactac ttactctggc ttcacggcaa
     1321 caattaatag actggcttga agcggataaa gttgcaggac cactactgcg ttcggcactt
     1381 cctgctggct ggtttattgc tgataaatct ggggcaggag agcgtggttc acggggtatc
     1441 attgccgcac ttggaccaga tggtaagcct tcccgtatcg tagttatcta cacgacgggt
     1501 agtcaggcaa ctatggacga acgaaataga cagattgctg aaataggggc ttcactgatt
     1561 aagcattggt aaaccgatac aattaaaggc tccttttgga gccttttttt ttggacggac
     1621 cggtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     1681 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     1741 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     1801 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     1861 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     1921 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     1981 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2041 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2101 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2161 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2221 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2281 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2341 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2401 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     2461 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     2521 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     2581 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     2641 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     2701 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     2761 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     2821 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     2881 ttcatggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     2941 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     3001 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg
     3061 tt
//
