LOCUS       sequence-69796-e        3395 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 3395)
  TITLE     Series-14: N-terminal Tag E.coli T7 Expression Vectors with Biobrick
            POLYPROMOTER Restriction Sites
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3395)
  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            "Unpublished"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 Thioredoxin TEV expression vector wit...
FEATURES             Location/Qualifiers
     source          1..3395
                     /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..624
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="SDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDEI
                     ADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFLD
                     ANLA"
     CDS             634..654
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     terminator      696..742
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1909..1948
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1979..2567
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2468..2487
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2721..2738
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2753..2892
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2818..2837)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2978..3000
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2994..3185)
                     /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 agcgataaaa ttattcacct gactgacgac agttttgaca cggatgtact caaagcggac
      361 ggggcgatcc tcgtcgattt ctgggcagag tggtgcggtc cgtgcaaaat gatcgccccg
      421 attctggatg aaatcgctga cgaatatcag ggcaaactga ccgttgcaaa actgaacatc
      481 gatcaaaacc ctggcactgc gccgaaatat ggcatccgtg gtatcccgac tctgctgctg
      541 ttcaaaaacg gtgaagtggc ggcaaccaaa gtgggtgcac tgtctaaagg tcagttgaaa
      601 gagttcctcg acgctaacct ggccgggatc gaggaaaacc tgtacttcca atccaatatt
      661 ggaagtggat aactggctgc tgccaccgct gagcatagca taaccccttg gggcctctaa
      721 acgggtcttg aggggttttt tgctgaaagg aggaactata tcaagtcccg aagtagctgt
      781 cagactgctg gcgaccgccg caactagtgg cgtcatgcgt gccgtatggt ctttcgcctg
      841 tagacgccct acaagccgta ctaaagtgga aagacccgca aaaggtatcg cgatcgccct
      901 gcagggaact atatccgact agtagcccgc ctaatgagcg ggcttttttt taattcccct
      961 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1021 taaatgcttc gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     1081 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctcgt gaaagtaaaa
     1141 gacgcagagg accaattggg ggcacgagtg ggatacatag aactggactt gaatagcggt
     1201 aaaatccttg agagttttcg ccctgaagag cgttttccaa tgatgagcac tttcaaagtt
     1261 ctgctatgtg gagcagtatt atcccgtgta gatgcggggc aagagcaact cggacgacga
     1321 atacactatt cgcagaatga cttggttgaa tactccccag tgacagaaaa gcaccttacg
     1381 gacggaatga cggtaagaga attatgtagt gccgccataa cgatgagtga taacactgcg
     1441 gcgaacttac ttctgacaac catcggtgga ccgaaggaat taaccgcttt tttgcacaat
     1501 atgggagacc atgtaactcg ccttgaccgt tgggaaccag aactgaatga agccatacca
     1561 aacgacgagc gagacaccac aatgcctgcg gcaatggcaa caacattacg caaactatta
     1621 actggcgaac tacttactct ggcttcacgg caacaattaa tagactggct tgaagcggat
     1681 aaagttgcag gaccactact gcgttcggca cttcctgctg gctggtttat tgctgataaa
     1741 tctggggcag gagagcgtgg ttcacggggt atcattgccg cacttggacc agatggtaag
     1801 ccttcccgta tcgtagttat ctacacgacg ggtagtcagg caactatgga cgaacgaaat
     1861 agacagattg ctgaaatagg ggcttcactg attaagcatt ggtaaaccga tacaattaaa
     1921 ggctcctttt ggagcctttt tttttggacg gaccggtaga aaagatcaaa ggatcttctt
     1981 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2041 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2101 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     2161 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2221 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2281 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2341 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2401 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2461 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2521 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2581 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     2641 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     2701 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     2761 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
     2821 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     2881 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     2941 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     3001 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     3061 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     3121 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     3181 gtcactgatg cctccgtgta agggggattt ctgttcatgg ggtaatgata ccgatgaaac
     3241 gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt
     3301 gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc
     3361 aatgccagcg cttcgttaat acagatgtag gtgtt
//
