LOCUS       sequence-462984-        3745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tetO-phi29 DNA polymerase 'dead' reporter: PT7-tetO-B0035-p2 
            D249E-T7 terminator, AmpR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3745)
  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 3745)
  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: pTE5427
FEATURES             Location/Qualifiers
     source          1..3745
                     /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"
     terminator      3684..3730
                     /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 gccgcgtaaa atgtacagct
     1981 gcgattttga aacgacgacg aaagttgaag attgccgtgt ctgggcctat ggttatatga
     2041 acatcgaaga ccattcagaa tataaaattg gcaactcgct ggatgaattt atggcgtggg
     2101 tgctgaaagt tcaggccgac ctgtacttcc acaatctgaa atttgatggt gcgttcatta
     2161 tcaactggct ggaacgtaat ggctttaaat ggagcgccga tggtctgccg aacacctata
     2221 atacgattat ctctcgtatg ggccaatggt atatgattga tatctgcctg ggctacaaag
     2281 gtaaacgcaa aattcatacc gtgatctatg acagcctgaa aaaactgccg tttccggtga
     2341 agaaaattgc gaaagatttc aaactgaccg tcctgaaagg cgatattgac tatcacaaag
     2401 aacgtccggt tggttacaaa atcacgccgg aagaatatgc gtacattaaa aacgatatcc
     2461 agattatcgc agaagctctg ctgattcagt ttaaacaagg cctggatcgc atgaccgccg
     2521 gcagtgactc cctgaaaggt ttcaaagata tcatcaccac gaaaaaattt aagaaagtgt
     2581 tcccgaccct gagcctgggt ctggataaag aagttcgtta tgcataccgc ggcggtttta
     2641 cgtggctgaa cgaccgtttc aaagaaaaag aaattggcga gggtatggtc tttgaagtga
     2701 atagtctgta tccggctcag atgtactccc gcctgctgcc gtatggcgaa ccgatcgttt
     2761 tcgagggtaa atatgtctgg gatgaagact acccgctgca tattcagcac atccgttgtg
     2821 aatttgaact gaaagaaggc tatattccga ccattcaaat caaacgtagc cgcttctata
     2881 agggtaacga atacctgaaa agctctggcg gtgaaatcgc agacctgtgg ctgagtaacg
     2941 tcgatctgga actgatgaaa gaacattacg atctgtacaa cgttgaatac atctccggcc
     3001 tgaaatttaa agccaccacg ggtctgttta aagatttcat tgacaaatgg acctacatca
     3061 aaaccacgtc tgaaggtgca atcaaacagc tggctaaact gatgctgaac agcctgtatg
     3121 gcaaatttgc atctaatccg gatgttaccg gtaaagtccc gtacctgaaa gaaaatggcg
     3181 ctctgggttt tcgcctgggc gaagaagaaa ccaaagatcc ggtgtatacg ccgatgggtg
     3241 ttttcattac cgcgtgggca cgttacacca ccatcaccgc cgcacaagcg tgctatgacc
     3301 gcattatcta ctgtgatacc gactcaattc atctgaccgg cacggaaatc ccggatgtga
     3361 ttaaagatat cgttgacccg aaaaaactgg gttattgggc acacgaatcg acctttaaac
     3421 gtgctaaata cctgcgccag aaaacgtaca tccaagacat ctacatgaaa gaagtcgatg
     3481 gcaaactggt ggaaggttca ccggatgact ataccgacat taaattttcg gtgaaatgcg
     3541 ccggcatgac cgataaaatt aagaaagaag tgacgttcga aaatttcaaa gtgggtttca
     3601 gtcgcaaaat gaaaccgaaa ccggtccaag ttccgggcgg cgttgtgctg gtcgatgaca
     3661 ccttcacgat caaataagct taatagcata accccttggg gcctctaaac gggtcttgag
     3721 gggttttttg cgctagacgg agacg
//
