LOCUS       sequence-414114-        3667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  p15A-tetR-tet-ME-RBS-mRFP-ME-U1-Barcode-U2-dbI terminator-oriT-Cm. 
            Plasmid backbone for fragment expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3667)
  AUTHORS   Biggs BW, Price MN, Lai D, Escobedo J, Fortanel Y, Huang YY, Kim K, 
            Trotter VV, Kuehl JV, Lui LM, Chakraborty R, Deutschbauer AM, Arkin 
            AP
  TITLE     High-throughput protein characterization by complementation using 
            DNA barcoded fragment libraries.
  JOURNAL   Mol Syst Biol. 2024 Oct 7. doi: 10.1038/s44320-024-00068-z.
  PUBMED    39375541
REFERENCE   2  (bases 1 to 3667)
  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: "Mol Syst 
            Biol. 2024 Oct 7. doi: 10.1038/s44320-024-00068-z."
COMMENT     Sequence Label: pBWB507
FEATURES             Location/Qualifiers
     source          1..3667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..627)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     532..551
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        643..698
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    679..697
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    714..732
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     CDS             753..1430
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     misc_feature    1431..1449
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     terminator      1562..1633
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1649..1676
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1702..1719)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1836..2381)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1916..1935)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            2466..2574
                     /note="incP origin of transfer"
     terminator      complement(2761..2855)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2876..3535)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3450..3469
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3536..3638)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 ttaagaccca ctttcacatt taagttgttt ttctaatccg catatgatca attcaaggcc
       61 gaataagaag gctggctctg caccttggtg atcaaataat tcgatagctt gtcgtaataa
      121 tggcggcata ctatcagtag taggtgtttc cctttcttct ttagcgactt gatgctcttg
      181 atcttccaat acgcaaccta aagtaaaatg ccccacagcg ctgagtgcat ataatgcatt
      241 ctctagtgaa aaaccttgtt ggcataaaaa ggctaattga ttttcgagag tttcatactg
      301 tttttctgta ggccgtgtac ctaaatgtac ttttgctcca tcgcgatgac ttagtaaagc
      361 acatctaaaa cttttagcgt tattacgtaa aaaatcttgc cagctttccc cttctaaagg
      421 gcaaaagtga gtatggtgcc tatctaacat ctcaatggct aaggcgtcga gcaaagcccg
      481 cttatttttt acatgccaat acaatgtagg ctgctctaca cctagcttct gggcgagttt
      541 acgggttgtt aaaccttcga ttccgacctc attaagcagc tctaatgcgc tgttaatcac
      601 tttactttta tctaatctag acatcattaa ttcctaattt ttgttgacac tctatcgttg
      661 atagagttat tttaccactc cctatcagtg atagagaaaa gaattcagac catagatgtg
      721 tataagagac agtttaagaa ggagatatac atatggcgag tagcgaagac gttatcaaag
      781 agttcatgcg tttcaaagtt cgtatggaag gttccgttaa cggtcacgag ttcgaaatcg
      841 aaggtgaagg tgaaggtcgt ccgtacgaag gtacccagac cgctaaactg aaagttacca
      901 aaggtggtcc gctgccgttc gcttgggaca tcctgtcccc gcagttccag tacggttcca
      961 aagcttacgt taaacacccg gctgacatcc cggactacct gaaactgtcc ttcccggaag
     1021 gtttcaaatg ggaacgtgtt atgaacttcg aagacggtgg tgttgttacc gttacccagg
     1081 actcctccct gcaagacggt gagttcatct acaaagttaa actgcgtggt accaacttcc
     1141 cgtccgacgg tccggttatg cagaaaaaaa ccatgggttg ggaagcttcc accgaacgta
     1201 tgtacccgga agacggtgct ctgaaaggtg aaatcaaaat gcgtctgaaa ctgaaagacg
     1261 gtggtcacta cgacgctgaa gttaaaacca cctacatggc taaaaaaccg gttcagctgc
     1321 cgggtgctta caaaaccgac atcaaactgg acatcacctc ccacaacgaa gactacacca
     1381 tcgttgaaca gtacgaacgt gctgaaggtc gtcactcaac cggtgcttaa ctgtctctta
     1441 tacacatctg tcgacctgca gcgtacgtaa catcgtccga tactgtaaga gacctcgtgg
     1501 acatcactgt ctagggataa cagggtaatg gatccaaact cgagtaagga tctccaggca
     1561 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     1621 ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta
     1681 taccgggata tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg
     1741 gcgagcggaa atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact
     1801 taacagggaa gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac
     1861 aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga
     1921 taccaggcgt ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt
     1981 accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc
     2041 cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg
     2101 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc
     2161 actggcagca gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt
     2221 aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt
     2281 tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt
     2341 tttcagagca agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc
     2401 agataaaata tttctagatt tcagtgcaat ttatctcttc aaatgtagca cctgttcttg
     2461 cttatcggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg cgaataaggg
     2521 acagtgaaga aggaacaccc gctcgcgggt gggcctactt cacctatcct gcccggctga
     2581 cgccgttgga tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt
     2641 gcgaaaaagg atggatatac cgaaaaaatc gctataatga ccccgaagca gggttatgca
     2701 gcggaaaaga atgtcatgat aagtcagccc catacgatat aagttgttac tagtgcttgg
     2761 attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag
     2821 ttctgaggtc attactggat ctatcaacag gagtccaagc gagctcgata tcaaattacg
     2881 ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga
     2941 agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt
     3001 gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt
     3061 ttaaatcaaa actggtgaaa ctcacccagg gattggctga aacgaaaaac atattctcaa
     3121 taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata
     3181 tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag
     3241 tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt
     3301 ctttcattgc catacgaaat tccggatgag cattcatcag gcgggcaaga atgtgaataa
     3361 aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca
     3421 gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt
     3481 tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag
     3541 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
     3601 cattatggtg aaagttggaa cctcttacgt gccgatcaag gtctcatttt cgccagatat
     3661 cgacgtc
//
