LOCUS       sequence-441879-        3139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Promoter probe vector with dropout sequence for promoter sequences 
            with 3WJdB reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3139)
  AUTHORS   Barthel S, Brenker L, Diehl C, Bohra N, Giaveri S, Paczia N, Erb TJ
  TITLE     In vitro transcription-based biosensing of glycolate for prototyping
            of a complex enzyme cascade.
  JOURNAL   Synth Biol (Oxf). 2024 Sep 20;9(1):ysae013. doi: 
            10.1093/synbio/ysae013. eCollection 2024.
  PUBMED    39399720
REFERENCE   2  (bases 1 to 3139)
  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: "Synth Biol 
            (Oxf)."; date: "2024-09-20"; volume: "9(1):ysae013. doi"; pages: " 
            10.1093/synbio/ysae013. eCollection 2024"
COMMENT     Sequence Label: pTE5417_p3WJdB_promoter dropout vector
FEATURES             Location/Qualifiers
     source          1..3139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             160..171
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             178..891
                     /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      903..974
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      990..1017
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1193..1239
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(1361..2152,2153..2221))
                     /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     1984..2003
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2222..2326)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2429..3017
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2918..2937
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggagtgagac cgaaagtgaa acgtgatttc atgcgtcatt ttgaacattt tgtaaatctt
       61 atttaataat gtgtgcggca attcacattt aatttatgaa tgttttctta acatcgcggc
      121 aactcaagaa acggcaggtt cggatcttag ctactagaga aagaggagaa atactagatg
      181 cgtaaaggcg aagagctgtt cactggtgtc gtccctattc tggtggaact ggatggtgat
      241 gtcaacggtc ataagttttc cgtgcgtggc gagggtgaag gtgacgcaac taatggtaaa
      301 ctgacgctga agttcatctg tactactggt aaactgccgg ttccttggcc gactctggta
      361 acgacgctga cttatggtgt tcagtgcttt gctcgttatc cggaccatat gaagcagcat
      421 gacttcttca agtccgccat gccggaaggc tatgtgcagg aacgcacgat ttcctttaag
      481 gatgacggca cgtacaaaac gcgtgcggaa gtgaaatttg aaggcgatac cctggtaaac
      541 cgcattgagc tgaaaggcat tgactttaaa gaggacggca atatcctggg ccataagctg
      601 gaatacaatt ttaacagcca caatgtttac atcaccgccg ataaacaaaa aaatggcatt
      661 aaagcgaatt ttaaaattcg ccacaacgtg gaggatggca gcgtgcagct ggctgatcac
      721 taccagcaaa acactccaat cggtgatggt cctgttctgc tgccagacaa tcactatctg
      781 agcacgcaaa gcgttctgtc taaagatccg aacgagaaac gcgatcatat ggttctgctg
      841 gagttcgtaa ccgcagcggg catcacgcat ggtatggatg aactgtacaa atgaccaggc
      901 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
      961 cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt
     1021 ataggtctca aatgcccaca tactctgatg atccgagacg gtcgggtcca gatattcgta
     1081 tctgtcgagt agagtgtggg ctcggatcat tcatggcaag agacggtcgg gtccagatat
     1141 tcgtatctgt cgagtagagt gtgggctctt gccatgtgta tgtggggctt aatagcataa
     1201 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc gcttgttgga gacgagctgc
     1261 ggccgcgatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1321 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1381 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1441 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1501 gggacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1561 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1621 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1681 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1741 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1801 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1861 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1921 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1981 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     2041 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     2101 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     2161 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2221 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     2281 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     2341 aagtgccacc tgtcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2401 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2461 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2521 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     2581 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2641 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2701 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2761 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2821 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2881 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2941 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3001 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3061 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtgcgg
     3121 ccgcaacacg tctcgagta
//
