LOCUS       sequence-476591-        3816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tetO-10xHis-phi29 DNA polymerase PovSak reporter: 
            PT7-tetO-B0035-10xHis-p2 povsak-T7 terminator, AmpR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3816)
  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 3816)
  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: pTE5451
FEATURES             Location/Qualifiers
     source          1..3816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(107..898,899..967))
                     /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     730..749
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(968..1072)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1175..1763
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1664..1683
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1890..1909
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1890..1908
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1911..1929
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1964..1993
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2006..2017
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      3755..3801
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 agctgcggcc gcgattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
       61 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
      121 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
      181 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
      241 taccgcggga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
      301 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
      361 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
      421 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
      481 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
      541 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
      601 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
      661 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
      721 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
      781 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
      841 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
      901 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
      961 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     1021 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     1081 cccgaaaagt gccacctgtc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     1141 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     1201 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     1261 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     1321 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     1381 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     1441 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     1501 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     1561 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     1621 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     1681 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     1741 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     1801 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     1861 gtgcggccgc aacacgtctc gtactggagt aatacgactc actataggga tccctatcag
     1921 tgatagagat actagagatt aaagaggaga ataatcaatg ggccatcatc atcatcatca
     1981 tcatcatcat cacagcagcg gccatatcga aggtcgtcat ggctctcgaa tgccgcgtaa
     2041 acgttacagc tgcgattttg aaacgacgac gaaagttgaa gattgccgtg tctgggccta
     2101 tggttatatg aacatcgaag accattcaga atataaaatt ggcaactcgc tggatgaatt
     2161 tatggcgtgg gcgctgaaag ttcaggccga cctgtacttc cacaatctga aatttgatgg
     2221 tgcgttcatt atcaactggc tggaacgtaa tggctttaaa tggagcgccg atggtctgcc
     2281 gaacacctat aatacgatta tctctcgtac cggccaatgg tatatgattg atatctgcct
     2341 gggctacaaa ggtaaacgca aaattcatac cgtgatctat gacagcctga aaaaactgcc
     2401 gtttccggtg aagaaaattg cgaaagattt caaactgacc gtcctgaaag gcgatattga
     2461 ctatcacaaa gaacgtccgg ttggttacaa aatcacgccg gaagaatatg cgtacattaa
     2521 aaacgatatc cagattatcg cagaagctct gctgattcag tttaaacaag gcctggatcg
     2581 catgaccgcc ggcagtgact ccctgaaaga tttcaaagat atcatcacca cgaaaaaatt
     2641 taagaaagtg ttcccgaccc tgagcctggg tctggataaa aaagttcgtt atgcataccg
     2701 cggcggtttt acgtggctga acgaccgttt caaagaaaaa gaaattggcg agggtatggt
     2761 ctttgatgtg aatagtctgt atccggctca gatgtactcc cgcctgctgc cgtatggcga
     2821 accgatcgtt ttcgagggta aatatgtctg ggatgaagac tacccgctgc atattcagca
     2881 catccgttgt gaatttgaac tgaaagaagg ctatattccg accattcaaa tcaaacgtag
     2941 ccgcttctat aagggtaacg aatacctgaa aagctctggc ggtgaaatcg cagacctgtg
     3001 gctgagtaac gtcgatctgg aactgatgaa agaacattac gatctgtaca acgttgaata
     3061 catctccggc ctgaaattta aagccaccac gggtctgttt aaagatttca ttgacaaatg
     3121 gacctacatc aaaaccacgt ctgaaggtgc aatcaaacag ctggctaaac tgatgctgaa
     3181 cagcctgtat ggcaaatttg catctaatcc ggatgttacc ggtaaagtcc cgtacctgaa
     3241 agaaaatggc gctctgggtt ttcgcctggg cgaagaagaa accaaagatc cggtgtatac
     3301 gccgatgggt gttttcatta ccgcgtgggc acgttacacc accatcaccg ccgcacaagc
     3361 gtgctatgac cgcattatct actgtgatac cgactcaatt catctgaccg gcacggaaat
     3421 cccggatgtg attaaagata tcgttgaccc gaaaaaactg ggttattggg cacacgaatc
     3481 gacctttaaa cgtgctaaat acctgcgcca gaaaacgtac atccaagaca tctacatgaa
     3541 agaagtcgat ggcaaactgg tggaaggttc accggatgac tataccgaca ttaaattttc
     3601 ggtgaaatgc gccggcatga ccgataaaat taagaaagaa gtgacgttcg aaaatttcaa
     3661 agtgggtttc agtcgcaaaa tgaccccgaa accggtccaa gttccgggcg gcgttgtgct
     3721 ggtcgataac accttcacga tcaaataagc ttaatagcat aaccccttgg ggcctctaaa
     3781 cgggtcttga ggggtttttt gcgctaatgg gagacg
//
