LOCUS       sequence-461677-        3592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone tet operator and TetR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3592)
  AUTHORS   Pignon E, Hollo G, Steiner T, van Vliet S, Schaerli Y
  TITLE     Uptake and leakage rates differentially shape community arrangement 
            and composition of microbial consortia.
  JOURNAL   ISME J. 2025 Jun 13:wraf122. doi: 10.1093/ismejo/wraf122.
  PUBMED    40509754
REFERENCE   2  (bases 1 to 3592)
  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: "ISME J. 
            2025 Jun 13:wraf122. doi: 10.1093/ismejo/wraf122."
COMMENT     Sequence Label: pEP28
FEATURES             Location/Qualifiers
     source          1..3592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             46..57
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             64..687
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /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="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(137..156)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      707..778
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      794..821
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    836..854
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    861..879
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      919..976
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     CDS             complement(1134..1793)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1708..1727
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1794..1896)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      complement(2491..3036)
                     /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(2571..2590)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3504..3522
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3526..3569)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    join(3572..3592,1)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 gtggtgcaaa acctttgcgg tatggcatga tagcgcctac tagagaaaga ggagaaatac
       61 tagatgtcca gattagataa aagtaaagtg attaacagcg cattagagct gcttaatgag
      121 gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt agagcagcct
      181 acattgtatt ggcatgtaaa aaataagcgg gctttgctcg acgccttagc cattgagatg
      241 ttagataggc accatactca cttttgccct ttagaagggg aaagctggca agatttttta
      301 cgtaataacg ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg agcaaaagta
      361 catttaggta cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt
      421 ttatgccaac aaggtttttc actagagaat gcattatatg cactcagcgc tgtggggcat
      481 tttactttag gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa
      541 acacctacta ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcac
      601 caaggtgcag agccagcctt cttattcggc cttgaattga tcatatgcgg attagaaaaa
      661 caacttaaat gtgaaagtgg gtcctaataa tactagagcc aggcatcaaa taaaacgaaa
      721 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta
      781 ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatatac tagagtccct
      841 atcagtgata gagattgaca tccctatcag tgatagagat actgagcact actagagaac
      901 tagtagcggc cgctgcagtc cggcaaaaaa gggcaaggtg tcaccaccct gccatttttc
      961 tttaaaaccg aaaagattac ttcgcgttat gcaggcttcc tcgctcactg actcgctgcg
     1021 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tctcgagtcc
     1081 cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc tgattacgcc
     1141 ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag
     1201 ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc
     1261 gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt
     1321 aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata
     1381 aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg
     1441 tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt
     1501 tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct
     1561 ttcattgcca tacggaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag
     1621 gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc
     1681 tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta
     1741 cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc cattttagct
     1801 tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca
     1861 ttatggtgaa agttggaacc tcttacgtgc cgatcaaaca ccccttgtat tactgtttat
     1921 gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca
     1981 gagattttga gacacaacgt ggctttgttg aataaatcga acttttgctg agttgaagga
     2041 tcagatcacg catcttcccg acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat
     2101 caccaactgg tccacctaca acaaagctct catcaaccgt ggctccctca ctttctggct
     2161 ggatgatggg gcgattcagg cctggtatga gtcagcaaca ccttcttcac gaggcagacc
     2221 tcagcgctag cggagtgtat actggcttac tatgttggca ctgatgaggg tgtcagtgaa
     2281 gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg tcagcagaat atgtgataca
     2341 ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag
     2401 cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca
     2461 gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca
     2521 tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca
     2581 ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg
     2641 tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt
     2701 aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg
     2761 ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg
     2821 cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc
     2881 taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa
     2941 gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca
     3001 gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat taaggggtct
     3061 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3121 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3181 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3241 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3301 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3361 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3421 actttatccg cctccatcca gtctattcca tggtgccacc tgacgtctaa gaaaccatta
     3481 ttatcatgac attaacctat aaaaataggc gtatcacgag gcagaatttc agataaaaaa
     3541 aatccttagc tttcgctaag gatgatttct ggaattcgcg gccgcttcta ga
//
