LOCUS       sequence-462966-        2797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sfGFP-Pepper reporter: PT7-tetO-B0035-sfGFP-pepper-T7 terminator, 
            AmpR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2797)
  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 2797)
  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: pTE5422
FEATURES             Location/Qualifiers
     source          1..2797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2..715
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      780..826
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(948..1739,1740..1808))
                     /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     1571..1590
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1809..1913)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2016..2604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2505..2524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2731..2750
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2731..2749
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2752..2770
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 aatgcgtaaa ggcgaagagc tgttcactgg tgtcgtccct attctggtgg aactggatgg
       61 tgatgtcaac ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg caactaatgg
      121 taaactgacg ctgaagttca tctgtactac tggtaaactg ccggttcctt ggccgactct
      181 ggtaacgacg ctgacttatg gtgttcagtg ctttgctcgt tatccggacc atatgaagca
      241 gcatgacttc ttcaagtccg ccatgccgga aggctatgtg caggaacgca cgatttcctt
      301 taaggatgac ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg ataccctggt
      361 aaaccgcatt gagctgaaag gcattgactt taaagaggac ggcaatatcc tgggccataa
      421 gctggaatac aattttaaca gccacaatgt ttacatcacc gccgataaac aaaaaaatgg
      481 cattaaagcg aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc agctggctga
      541 tcactaccag caaaacactc caatcggtga tggtcctgtt ctgctgccag acaatcacta
      601 tctgagcacg caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc atatggttct
      661 gctggagttc gtaaccgcag cgggcatcac gcatggtatg gatgaactgt acaaataagc
      721 ttggccccaa tcgtggcgtg tcggcctgct tcggcaggca ctggcgccgg gccgcttaat
      781 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgcgct tgttggagac
      841 gagctgcggc cgcgattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
      901 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
      961 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     1021 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     1081 ataccgcggg acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     1141 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     1201 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     1261 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     1321 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     1381 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     1441 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     1501 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1561 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     1621 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1681 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1741 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1801 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     1861 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     1921 ccccgaaaag tgccacctgt catgaccaaa atcccttaac gtgagttttc gttccactga
     1981 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2041 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2101 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2161 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2221 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2281 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2341 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2401 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2461 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2521 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2581 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     2641 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     2701 cgtgcggccg caacacgtct cgagtaggag taatacgact cactataggg atccctatca
     2761 gtgatagaga tactagagat taaagaggag aataatc
//
