LOCUS       sequence-462976-        3817 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inactive tetO-10xHis-phi29 DNA polymerase wild-type reporter: 
            PT7-tetO-B0035-10xHis-p2 dead-T7 terminator, AmpR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3817)
  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 3817)
  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: pTE5452
FEATURES             Location/Qualifiers
     source          1..3817
                     /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    1912..1930
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1965..1994
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2007..2018
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      3756..3802
                     /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 cgtactggag taatacgact cactataggg atccctatca
     1921 gtgatagaga tactagagat taaagaggag aataatcaat gggccatcat catcatcatc
     1981 atcatcatca tcacagcagc ggccatatcg aaggtcgtca tggctctcga atgccgcgta
     2041 aaatgtacag ctgcgatttt gaaacgacga cgaaagttga agattgccgt gtctgggcct
     2101 atggttatat gaacatcgaa gaccattcag aatataaaat tggcaactcg ctggatgaat
     2161 ttatggcgtg ggtgctgaaa gttcaggccg acctgtactt ccacaatctg aaatttgatg
     2221 gtgcgttcat tatcaactgg ctggaacgta atggctttaa atggagcgcc gatggtctgc
     2281 cgaacaccta taatacgatt atctctcgta tgggccaatg gtatatgatt gatatctgcc
     2341 tgggctacaa aggtaaacgc aaaattcata ccgtgatcta tgacagcctg aaaaaactgc
     2401 cgtttccggt gaagaaaatt gcgaaagatt tcaaactgac cgtcctgaaa ggcgatattg
     2461 actatcacaa agaacgtccg gttggttaca aaatcacgcc ggaagaatat gcgtacatta
     2521 aaaacgatat ccagattatc gcagaagctc tgctgattca gtttaaacaa ggcctggatc
     2581 gcatgaccgc cggcagtgac tccctgaaag gtttcaaaga tatcatcacc acgaaaaaat
     2641 ttaagaaagt gttcccgacc ctgagcctgg gtctggataa agaagttcgt tatgcatacc
     2701 gcggcggttt tacgtggctg aacgaccgtt tcaaagaaaa agaaattggc gagggtatgg
     2761 tctttgaagt gaatagtctg tatccggctc agatgtactc ccgcctgctg ccgtatggcg
     2821 aaccgatcgt tttcgagggt aaatatgtct gggatgaaga ctacccgctg catattcagc
     2881 acatccgttg tgaatttgaa ctgaaagaag gctatattcc gaccattcaa atcaaacgta
     2941 gccgcttcta taagggtaac gaatacctga aaagctctgg cggtgaaatc gcagacctgt
     3001 ggctgagtaa cgtcgatctg gaactgatga aagaacatta cgatctgtac aacgttgaat
     3061 acatctccgg cctgaaattt aaagccacca cgggtctgtt taaagatttc attgacaaat
     3121 ggacctacat caaaaccacg tctgaaggtg caatcaaaca gctggctaaa ctgatgctga
     3181 acagcctgta tggcaaattt gcatctaatc cggatgttac cggtaaagtc ccgtacctga
     3241 aagaaaatgg cgctctgggt tttcgcctgg gcgaagaaga aaccaaagat ccggtgtata
     3301 cgccgatggg tgttttcatt accgcgtggg cacgttacac caccatcacc gccgcacaag
     3361 cgtgctatga ccgcattatc tactgtgata ccgactcaat tcatctgacc ggcacggaaa
     3421 tcccggatgt gattaaagat atcgttgacc cgaaaaaact gggttattgg gcacacgaat
     3481 cgacctttaa acgtgctaaa tacctgcgcc agaaaacgta catccaagac atctacatga
     3541 aagaagtcga tggcaaactg gtggaaggtt caccggatga ctataccgac attaaatttt
     3601 cggtgaaatg cgccggcatg accgataaaa ttaagaaaga agtgacgttc gaaaatttca
     3661 aagtgggttt cagtcgcaaa atgaaaccga aaccggtcca agttccgggc ggcgttgtgc
     3721 tggtcgatga caccttcacg atcaaataag cttaatagca taaccccttg gggcctctaa
     3781 acgggtcttg aggggttttt tgcgctagac ggagacg
//
