LOCUS       sequence-482107-        4042 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Over-express E. coli Pol III subunit theta in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4042)
  TITLE     O'Donnell lab plasmids 2025
REFERENCE   2  (bases 1 to 4042)
  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"
COMMENT     Sequence Label: Ec holE-pCDFDuet
FEATURES             Location/Qualifiers
     source          1..4042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(30..74)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     protein_bind    183..207
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(208..226)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    609..633
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(634..652)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        698..775
                     /gene="lacI"
                     /label=lacI promoter
     CDS             776..1858
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(795..814)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        2919..3010
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3011..3802
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     terminator      complement(3973..4020)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4016..4034
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 gcctaggtta attaagctgc gctagtagac gagtccatgt gctggcgttc aaatttcgca
       61 gcagcggttt ctttaccaga ctcgagggta ccgacgtcag cgatcgcgtg gccggccgat
      121 atccaattga gatctgccat atgtatatct ccttcttata cttaactaat atactaagat
      181 ggggaattgt tatccgctca caattcccct atagtgagtc gtattaattt cgattatgcg
      241 gccgtgtaca atacgattac tttctgttcg acttaagcat tatgcggccg caagcttgtc
      301 gacctgcagg cgcgccgagc tcgaattcgg atccttattt aagtttgggc tcgtaaggta
      361 aacgtgacag attgaccgaa gccaaacggt gggcaataag ccgctcgcga aaccagctgc
      421 gcaaatgttc aggctgttca cgttcaaccg cttcagcgat caccggcata ttgtagcgtt
      481 ctttaaatgc caccccggcc gccgccaaat cgacattcac tttatccatt tctgtttgat
      541 ccagtttagc cagattcttc aacatggtat atctccttat taaagttaaa caaaattatt
      601 tctacagggg aattgttatc cgctcacaat tcccctatag tgagtcgtat taatttccta
      661 atgcaggagt cgcataaggg agagcgtcga gatcccggac accatcgaat ggcgcaaaac
      721 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa
      781 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg
      841 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat
      901 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt
      961 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc
     1021 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag
     1081 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct
     1141 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
     1201 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
     1261 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
     1321 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
     1381 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
     1441 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
     1501 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
     1561 gcgcgttggt gcggacatct cggtagtggg atacgacgat accgaagaca gctcatgtta
     1621 tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
     1681 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
     1741 actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt
     1801 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     1861 gcaacgcaat taatgtaagt tagctcactc attaggcacc gggatctcga ccgatgccct
     1921 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact aacatgagaa
     1981 ttacaactta tatcgtatgg ggctgacttc aggtgctaca tttgaagaga taaattgcac
     2041 tgaaatctag agcggttcag tagaaaagat caaaggatct tcttgagatc ctttttttct
     2101 gcgcgtaatc ttttgccctg taaacgaaaa aaccacctgg ggaggtggtt tgatcgaagg
     2161 ttaagcagtt ccccaactga cttaaccttc gaaaaaacca cctggggagg tggtttgatc
     2221 gaaggttaag tcagttgggg aactgcttaa ccgtggtaac tggctttcgc agagcacagc
     2281 aaccaaatct gtccttccag tgtagccgga ctttggcgca cacttcaaga gcaaccgcgt
     2341 gtttagctaa acaaatcctc tgcgaactcc cagttaccaa tggctgctgc cagtggcgtt
     2401 ttaccgtgct tttccgggtt ggactcaagt gaacagttac cggataaggc gcagcagtcg
     2461 ggctgaacgg ggagttcttg cttacagccc agcttggagc gaacgaccta caccgagccg
     2521 agataccagt gtgtgagcta tgagaaagcg ccacacttcc cgtaagggag aaaggcggaa
     2581 caggtatccg gtaaacggca gggtcggaac aggagagcgc aagagggagc gacccgccgg
     2641 aaacggtggg gatctttaag tcctgtcggg tttcgcccgt actgtcagat tcatggttga
     2701 gcctcacggc tcccacagat gcaccggaaa agcgtctgtt tatgtgaact ctggcaggag
     2761 ggcggagcct atggaaaaac gccaccggcg cggccctgct gttttgcctc acatgttagt
     2821 cccctgctta tccacggaat ctgtgggtaa ctttgtatgt gtccgcagcg cccgccgcag
     2881 tctcacgccc ggagcgtagc gaccgagtga gctagctatt tgtttatttt tctaaataca
     2941 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3001 aaggaagagt atgagggaag cggtgatcgc cgaagtatcg actcaactat cagaggtagt
     3061 tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt acggctccgc
     3121 agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg tgaccgtaag
     3181 gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt cggcttcccc
     3241 tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg acgacatcat
     3301 tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc gcaatgacat
     3361 tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct tgctgacaaa
     3421 agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct ttgatccggt
     3481 tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga actcgccgcc
     3541 cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc
     3601 agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc
     3661 ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac aagaagaaga
     3721 tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga aaggcgagat
     3781 caccaaggta gtcggcaaat aatgtctaac aattcgttca agccgagggg ccgcaagatc
     3841 cggccacgat gacccggtcg tcggttcagg gcagggtcgt taaatagccg cttatgtcta
     3901 ttgctggttt accggtttat tgactaccgg aagcagtgtg accgtgtgct tctcaaatgc
     3961 ctgaggtttc agcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctag
     4021 ttattgctca gcggtggcag ca
//
