LOCUS       sequence-462971-        3744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tetO-phi29 DNA polymerase wild-type reporter: PT7-tetO-B0035-p2 
            wt-T7 terminator, AmpR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3744)
  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 3744)
  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: pTE5426
FEATURES             Location/Qualifiers
     source          1..3744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1727..1773
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(1895..2686,2687..2755))
                     /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     2518..2537
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2756..2860)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2963..3551
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3452..3471
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3678..3697
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3678..3696
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3699..3717
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 aatgccgcgt aaaatgtaca gctgcgattt tgaaacgacg acgaaagttg aagattgccg
       61 tgtctgggcc tatggttata tgaacatcga agaccattca gaatataaaa ttggcaactc
      121 gctggatgaa tttatggcgt gggtgctgaa agttcaggcc gacctgtact tccacaatct
      181 gaaatttgat ggtgcgttca ttatcaactg gctggaacgt aatggcttta aatggagcgc
      241 cgatggtctg ccgaacacct ataatacgat tatctctcgt atgggccaat ggtatatgat
      301 tgatatctgc ctgggctaca aaggtaaacg caaaattcat accgtgatct atgacagcct
      361 gaaaaaactg ccgtttccgg tgaagaaaat tgcgaaagat ttcaaactga ccgtcctgaa
      421 aggcgatatt gactatcaca aagaacgtcc ggttggttac aaaatcacgc cggaagaata
      481 tgcgtacatt aaaaacgata tccagattat cgcagaagct ctgctgattc agtttaaaca
      541 aggcctggat cgcatgaccg ccggcagtga ctccctgaaa ggtttcaaag atatcatcac
      601 cacgaaaaaa tttaagaaag tgttcccgac cctgagcctg ggtctggata aagaagttcg
      661 ttatgcatac cgcggcggtt ttacgtggct gaacgaccgt ttcaaagaaa aagaaattgg
      721 cgagggtatg gtctttgatg tgaatagtct gtatccggct cagatgtact cccgcctgct
      781 gccgtatggc gaaccgatcg ttttcgaggg taaatatgtc tgggatgaag actacccgct
      841 gcatattcag cacatccgtt gtgaatttga actgaaagaa ggctatattc cgaccattca
      901 aatcaaacgt agccgcttct ataagggtaa cgaatacctg aaaagctctg gcggtgaaat
      961 cgcagacctg tggctgagta acgtcgatct ggaactgatg aaagaacatt acgatctgta
     1021 caacgttgaa tacatctccg gcctgaaatt taaagccacc acgggtctgt ttaaagattt
     1081 cattgacaaa tggacctaca tcaaaaccac gtctgaaggt gcaatcaaac agctggctaa
     1141 actgatgctg aacagcctgt atggcaaatt tgcatctaat ccggatgtta ccggtaaagt
     1201 cccgtacctg aaagaaaatg gcgctctggg ttttcgcctg ggcgaagaag aaaccaaaga
     1261 tccggtgtat acgccgatgg gtgttttcat taccgcgtgg gcacgttaca ccaccatcac
     1321 cgccgcacaa gcgtgctatg accgcattat ctactgtgat accgactcaa ttcatctgac
     1381 cggcacggaa atcccggatg tgattaaaga tatcgttgac ccgaaaaaac tgggttattg
     1441 ggcacacgaa tcgaccttta aacgtgctaa atacctgcgc cagaaaacgt acatccaaga
     1501 catctacatg aaagaagtcg atggcaaact ggtggaaggt tcaccggatg actataccga
     1561 cattaaattt tcggtgaaat gcgccggcat gaccgataaa attaagaaag aagtgacgtt
     1621 cgaaaatttc aaagtgggtt tcagtcgcaa aatgaaaccg aaaccggtcc aagttccggg
     1681 cggcgttgtg ctggtcgatg acaccttcac gatcaaataa gcttaatagc ataacccctt
     1741 ggggcctcta aacgggtctt gaggggtttt ttgcgctaat gggagacgag ctgcggccgc
     1801 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     1861 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     1921 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     1981 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgggacc
     2041 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     2101 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     2161 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     2221 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     2281 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     2341 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     2401 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     2461 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     2521 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     2581 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     2641 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     2701 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     2761 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     2821 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     2881 cacctgtcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2941 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3001 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3061 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     3121 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3181 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3241 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3301 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3361 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3421 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3481 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3541 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3601 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt gcggccgcaa
     3661 cacgtctcgt actggagtaa tacgactcac tatagggatc cctatcagtg atagagatac
     3721 tagagattaa agaggagaat aatc
//
