LOCUS       sequence-461676-        5126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PutP (Sodium/proline symporter) under tet operator, TetR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5126)
  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 5126)
  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: pEP17
FEATURES             Location/Qualifiers
     source          1..5126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             45..56
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             63..686
                     /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(136..155)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      706..777
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      793..820
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    835..853
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    860..878
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             897..908
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2452..2509
                     /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(2667..3326)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3241..3260
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3327..3429)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      complement(4024..4569)
                     /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(4104..4123)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5037..5055
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(5059..5102)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    5105..5126
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 tggtgcaaaa cctttgcggt atggcatgat agcgcctact agagaaagag gagaaatact
       61 agatgtccag attagataaa agtaaagtga ttaacagcgc attagagctg cttaatgagg
      121 tcggaatcga aggtttaaca acccgtaaac tcgcccagaa gctaggtgta gagcagccta
      181 cattgtattg gcatgtaaaa aataagcggg ctttgctcga cgccttagcc attgagatgt
      241 tagataggca ccatactcac ttttgccctt tagaagggga aagctggcaa gattttttac
      301 gtaataacgc taaaagtttt agatgtgctt tactaagtca tcgcgatgga gcaaaagtac
      361 atttaggtac acggcctaca gaaaaacagt atgaaactct cgaaaatcaa ttagcctttt
      421 tatgccaaca aggtttttca ctagagaatg cattatatgc actcagcgct gtggggcatt
      481 ttactttagg ttgcgtattg gaagatcaag agcatcaagt cgctaaagaa gaaagggaaa
      541 cacctactac tgatagtatg ccgccattat tacgacaagc tatcgaatta tttgatcacc
      601 aaggtgcaga gccagccttc ttattcggcc ttgaattgat catatgcgga ttagaaaaac
      661 aacttaaatg tgaaagtggg tcctaataat actagagcca ggcatcaaat aaaacgaaag
      721 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac
      781 tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttatatact agagtcccta
      841 tcagtgatag agattgacat ccctatcagt gatagagata ctgagcacta ctagagaaag
      901 aggagaaata ctagatggct attagcacac cgatgttggt gacattttgt gtctatatct
      961 ttggcatgat attgattggg tttatcgcct ggcgatcaac gaaaaacttt gacgactata
     1021 ttctgggcgg tcgtagtctt gggccattcg tgacggcatt atcggcgggt gcgtcggata
     1081 tgagcggctg gctgttaatg gggttgccgg gcgctgtttt tctttccggg atttccgaaa
     1141 gctggatcgc cattggcctg acattaggcg cgtggattaa ctggaagctg gtggccgggc
     1201 ggttgcgtgt gcataccgaa tacaacaata acgccttaac actgccggat tatttcaccg
     1261 ggcgctttga agataaaagc cgcattttgc gcattatctc tgcgctggtt attttgctgt
     1321 tcttcaccat ttattgcgct tcgggcattg tggcaggcgc gcgtctgttt gaaagtacct
     1381 ttggcatgag ctacgaaacg gctctgtggg cgggcgctgc ggcgacgatc ctttacacct
     1441 ttattggcgg tttcctcgcg gtgagctgga ctgacactgt acaggccagc ctgatgattt
     1501 ttgccctgat cctgacgccg gttatcgtca ttatcagtgt cggtggcttt ggtgactcgc
     1561 tggaagtgat caaacaaaag agcatcgaaa acgttgatat gctcaaaggt ctgaactttg
     1621 ttgccattat ctcactgatg ggttgggggc tgggttactt cgggcagccg cacattctgg
     1681 cgcgttttat ggcggcggat tctcaccaca gcattgtcca tgcgcgtcgt attagtatga
     1741 cctggatgat cctctgcctg gcaggggcgg tggctgtcgg cttctttggg attgcttact
     1801 ttaacgatca tccggcgttg gctggtgcgg taaatcagaa cgccgagcgt gtgtttatcg
     1861 aactggcgca aattctgttt aacccgtgga ttgccgggat tctgctgtcg gcaattctgg
     1921 cggcggtaat gtcaacctta agttgccagc tgctggtgtg ctccagtgcg attaccgaag
     1981 atttgtacaa agcgtttctg cgtaaacatg ccagccagaa agagctggtg tgggtagggc
     2041 gtgtgatggt gctggtggtg gcgctggtgg cgattgcgct ggcggcaaac ccggaaaacc
     2101 gcgtgctggg cttagtgagc tacgcgtggg caggctttgg cgcggcgttt ggtccagtgg
     2161 tgctgttctc ggtgatgtgg tcacgcatga cgcgtaacgg tgcgctggcg gggatgatca
     2221 tcggtgcgct gacggttatc gtctggaaac agttcggctg gctgggactg tacgaaatta
     2281 ttccgggctt tatcttcggc agtattggga ttgtagtgtt tagtttgctg ggtaaagcgc
     2341 cgtcagcggc gatgcaaaaa cgctttgccg aggccgatgc gcactatcat tcggctccgc
     2401 cgtcacggtt gcaggaaagc taagggactt aactagtagc ggccgctgca gtccggcaaa
     2461 aaagggcaag gtgtcaccac cctgccattt ttctttaaaa ccgaaaagat tacttcgcgt
     2521 tatgcaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2581 atcagctcac tcaaaggcgg taatctcgag tcccgtcaag tcagcgtaat gctctgccag
     2641 tgttacaacc aattaaccaa ttctgattac gccccgccct gccactcatc gcagtactgt
     2701 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga
     2761 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg
     2821 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag
     2881 ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt
     2941 tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg
     3001 tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg
     3061 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacggaa ttccggatga
     3121 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc
     3181 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
     3241 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
     3301 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac
     3361 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg
     3421 tgccgatcaa acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     3481 tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa cgtggctttg
     3541 ttgaataaat cgaacttttg ctgagttgaa ggatcagatc acgcatcttc ccgacaacgc
     3601 agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac tggtccacct acaacaaagc
     3661 tctcatcaac cgtggctccc tcactttctg gctggatgat ggggcgattc aggcctggta
     3721 tgagtcagca acaccttctt cacgaggcag acctcagcgc tagcggagtg tatactggct
     3781 tactatgttg gcactgatga gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg
     3841 ctgcaccggt gcgtcagcag aatatgtgat acaggatata ttccgcttcc tcgctcactg
     3901 actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag
     3961 atttcctgga agatgccagg aagatactta acagggaagt gagagggccg cggcaaagcc
     4021 gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg
     4081 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg gctccctcgt
     4141 gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt
     4201 gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta
     4261 tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4321 ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag
     4381 gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac
     4441 tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa
     4501 aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa
     4561 cgatctcaag aagatcatct tattaagggg tctgacgctc agtggaacga aaactcacgt
     4621 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4681 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4741 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4801 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4861 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4921 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4981 ccatggtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     5041 ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt agctttcgct aaggatgatt
     5101 tctggaattc gcggccgctt ctagag
//
