LOCUS       sequence-462986-        2652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for TetR: PT7-lacO-B0035-TetR-T7 terminator, AmpR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2652)
  AUTHORS   Barthel S, Hoffmann-Becking M, Karimov IG, Erb TJ
  TITLE     Genetically encoded control of in vitro transcription-translation 
            coupled DNA replication
  JOURNAL   bioRxiv 2025.07.08.663768; doi: 10.1101/2025.07.08.663768
REFERENCE   2  (bases 1 to 2652)
  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: "bioRxiv 
            2025.07.08.663768; doi: 10.1101/2025.07.08.663768"
COMMENT     Sequence Label: pTE5428
FEATURES             Location/Qualifiers
     source          1..2652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(108..899,900..968))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     731..750
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(969..1073)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1176..1764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1665..1684
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1891..1910
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1891..1909
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1914..1930
                     /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)."
     CDS             1961..2584
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(2034..2053)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2591..2637
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 agactgcggc cgcgattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
       61 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
      121 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
      181 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
      241 ataccgcggg acccacgctc accggctcca gatttatcag caataaacca gccagccgga
      301 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
      361 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
      421 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
      481 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
      541 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
      601 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
      661 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
      721 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
      781 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
      841 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
      901 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
      961 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     1021 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     1081 ccccgaaaag tgccacctgt catgaccaaa atcccttaac gtgagttttc gttccactga
     1141 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1201 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1261 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1321 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1381 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1441 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     1501 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     1561 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     1621 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     1681 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     1741 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     1801 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     1861 cgtgcggccg caacacgtct cgagtaggag taatacgact cactataggg gaattgtgag
     1921 cggataacaa tttactagag attaaagagg agaataatca atgtctagat tagataaaag
     1981 taaagtgatt aacagcgcat tagagctgct taatgaggtc ggaatcgaag gtttaacaac
     2041 ccgtaaactc gcccagaagc taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa
     2101 taagcgggct ttgctcgacg ccttagccat tgagatgtta gataggcacc atactcactt
     2161 ttgcccttta gaaggggaaa gctggcaaga ttttttacgt aataacgcta aaagttttag
     2221 atgtgcttta ctaagtcatc gcgatggagc aaaagtacat ttaggtacac ggcctacaga
     2281 aaaacagtat gaaactctcg aaaatcaatt agccttttta tgccaacaag gtttttcact
     2341 agagaatgca ttatatgcac tcagcgctgt ggggcatttt actttaggtt gcgtattgga
     2401 agatcaagag catcaagtcg ctaaagaaga aagggaaaca cctactactg atagtatgcc
     2461 gccattatta cgacaagcta tcgaattatt tgatcaccaa ggtgcagagc cagccttctt
     2521 attcggcctt gaattgatca tatgcggatt agagaagcaa cttaagtgtg aaagtgggtc
     2581 ttaagcttaa tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgcgc
     2641 ttgttggaga cg
//
