LOCUS       sequence-348284-        5543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet inducible, KAN Resistant, non labelled synthesis domain of the 
            E. coli relA gene on a low copy backbone. Used to induce 
            controllable synthesis of ppGpp in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5543)
  AUTHORS   Buke F, Grilli J, Cosentino Lagomarsino M, Bokinsky G, Tans SJ
  TITLE     ppGpp is a bacterial cell size regulator.
  JOURNAL   Curr Biol. 2022 Feb 28;32(4):870-877.e5. doi: 
            10.1016/j.cub.2021.12.033. Epub 2022 Jan 5.
  PUBMED    34990598
REFERENCE   2  (bases 1 to 5543)
  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"; doi: 
            "10.1016/j.cub.2021.12.033"; journalName: "Curr Biol"; date: 
            "2022-02-28- 28"; volume: "32"; issue: "4"; pages: "870-877.e5"
COMMENT     Sequence Label: pRelA'
FEATURES             Location/Qualifiers
     source          1..5543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(172..266)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(297..1091)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(408..427)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1018..1037
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(1234..1860)
                     /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     1765..1784
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        1876..1931
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    1912..1930
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      3358..3429
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3445..3472
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4003..4953)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 ctgctatgtg gtgctatctg actttttgct gttcagcagt tcctgccctc tgattttcca
       61 gtctgaccac ttcggattat cccgtgacag gtcattcaga ctggctaatg cacccagtaa
      121 ggcagcggta tcatcaacag gcttacccgt cttactgtcc ctagtgcttg gattctcacc
      181 aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt
      241 cattactgga tctatcaaca ggagtccaag cgagctctcg aaccccagag tcccgctcag
      301 aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg
      361 taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta
      421 gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca
      481 gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg
      541 agatcctcgc cgtcgggcat gcgcgccttg agcctggcga acagttcggc tggcgcgagc
      601 ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt
      661 gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta
      721 tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat
      781 gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg
      841 acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct
      901 gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg
      961 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc
     1021 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct
     1081 tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cagatcatga tcccctgcgc
     1141 catcagatcc ttggcggcaa gaaagccatc cagtttactt tgcagggctt cccaacctta
     1201 ccagagggcg ccccagctgg caattccgac gtcttaagac ccactttcac atttaagttg
     1261 tttttctaat ccgcatatga tcaattcaag gccgaataag aaggctggct ctgcaccttg
     1321 gtgatcaaat aattcgatag cttgtcgtaa taatggcggc atactatcag tagtaggtgt
     1381 ttccctttct tctttagcga cttgatgctc ttgatcttcc aatacgcaac ctaaagtaaa
     1441 atgccccaca gcgctgagtg catataatgc attctctagt gaaaaacctt gttggcataa
     1501 aaaggctaat tgattttcga gagtttcata ctgtttttct gtaggccgtg tacctaaatg
     1561 tacttttgct ccatcgcgat gacttagtaa agcacatcta aaacttttag cgttattacg
     1621 taaaaaatct tgccagcttt ccccttctaa agggcaaaag tgagtatggt gcctatctaa
     1681 catctcaatg gctaaggcgt cgagcaaagc ccgcttattt tttacatgcc aatacaatgt
     1741 aggctgctct acacctagct tctgggcgag tttacgggtt gttaaacctt cgattccgac
     1801 ctcattaagc agctctaatg cgctgttaat cactttactt ttatctaatc tagacatcat
     1861 taattcctaa tttttgttga cactctatcg ttgatagagt tattttacca ctccctatca
     1921 gtgatagaga aaagaattca aaagatctag gaggaaaaaa atggttgcgg taagaagtgc
     1981 acatatcaat aaggctggtg aatttgatcc ggaaaaatgg atcgcaagtc tgggtattac
     2041 cagccagaag tcgtgtgagt gcttagccga aacctgggcg tattgtctgc aacagacgca
     2101 ggggcatccg gatgccagtc tgttattgtg gcgtggtgtt gagatggtgg agatcctctc
     2161 gacattaagt atggacattg acacgctgcg ggcggcgctg cttttccctc tggcggatgc
     2221 caacgtagtc agcgaagatg tgctgcgtga gagcgtcggt aagtcggtcg ttaaccttat
     2281 tcacggcgtg cgtgatatgg cggcgatccg ccagctgaaa gcgacgcaca ctgattctgt
     2341 ttcctccgaa caggtcgata acgttcgccg gatgttattg gcgatggtcg atgattttcg
     2401 ctgcgtagtc atcaaactgg cggagcgtat tgctcatctg cgcgaagtaa aagatgcgcc
     2461 ggaagatgaa cgtgtactgg cggcaaaaga gtgtaccaac atctacgcac cgctggctaa
     2521 ccgtctcgga atcggacaac tgaaatggga actggaagat tactgcttcc gttacctcca
     2581 tccaaccgaa tacaaacgaa ttgccaaact gctgcatgaa cggcgtctcg accgcgaaca
     2641 ctacatcgaa gagttcgttg gtcatctgcg cgctgagatg aaagctgaag gcgttaaagc
     2701 ggaagtgtat ggtcgtccga aacacatcta cagcatctgg cgtaaaatgc agaaaaagaa
     2761 cctcgccttt gatgagctgt ttgatgtgcg tgcggtacgt attgtcgccg agcgtttaca
     2821 ggattgctat gccgcactgg ggatagtgca cactcactat cgccacctgc cggatgagtt
     2881 tgacgattac gtcgctaacc cgaaaccaaa cggttatcag tctattcata ccgtggttct
     2941 ggggccgggt ggaaaaaccg ttgagatcca aatccgcacc aaacagatgc atgaagatgc
     3001 agagttgggt gttgctgcgc actggaaata taaagagggc gcggctgctg gcggcgcacg
     3061 ttcgggacat gaagaccgga ttgcctggct gcgtaaactg attgcgtggc aggaagagat
     3121 ggctgattcc ggcgaaatgc tcgacgaagt acgtagtcag gtctttgacg accgggtgta
     3181 cgtctttacg ccgaaaggtg atgtcgttga tttgcctgcg ggatcaacgc cgctggactt
     3241 cgcttaccac atccacagtg atgtcggaca ccgctgcatc ggggcaaaaa ttggcgggcg
     3301 cattgtgccg ttcacctacc agctgggatc ctaactcgag taaggatctc caggcatcaa
     3361 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     3421 aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatacc
     3481 tagggtacgg gttttgctgc ccgcaaacgg gctgttctgg tgttgctagt ttgttatcag
     3541 aatcgcagat ccggcttcag ccggtttgcc ggctgaaagc gctatttctt ccagaattgc
     3601 catgattttt tccccacggg aggcgtcact ggctcccgtg ttgtcggcag ctttgattcg
     3661 ataagcagca tcgcctgttt caggctgtct atgtgtgact gttgagctgt aacaagttgt
     3721 ctcaggtgtt caatttcatg ttctagttgc tttgttttac tggtttcacc tgttctatta
     3781 ggtgttacat gctgttcatc tgttacattg tcgatctgtt catggtgaac agctttgaat
     3841 gcaccaaaaa ctcgtaaaag ctctgatgta tctatctttt ttacaccgtt ttcatctgtg
     3901 catatggaca gttttccctt tgatatgtaa cggtgaacag ttgttctact tttgtttgtt
     3961 agtcttgatg cttcactgat agatacaaga gccataagaa cctcagatcc ttccgtattt
     4021 agccagtatg ttctctagtg tggttcgttg tttttgcgtg agccatgaga acgaaccatt
     4081 gagatcatac ttactttgca tgtcactcaa aaattttgcc tcaaaactgg tgagctgaat
     4141 ttttgcagtt aaagcatcgt gtagtgtttt tcttagtccg ttatgtaggt aggaatctga
     4201 tgtaatggtt gttggtattt tgtcaccatt catttttatc tggttgttct caagttcggt
     4261 tacgagatcc atttgtctat ctagttcaac ttggaaaatc aacgtatcag tcgggcggcc
     4321 tcgcttatca accaccaatt tcatattgct gtaagtgttt aaatctttac ttattggttt
     4381 caaaacccat tggttaagcc ttttaaactc atggtagtta ttttcaagca ttaacatgaa
     4441 cttaaattca tcaaggctaa tctctatatt tgccttgtga gttttctttt gtgttagttc
     4501 ttttaataac cactcataaa tcctcataga gtatttgttt tcaaaagact taacatgttc
     4561 cagattatat tttatgaatt tttttaactg gaaaagataa ggcaatatct cttcactaaa
     4621 aactaattct aatttttcgc ttgagaactt ggcatagttt gtccactgga aaatctcaaa
     4681 gcctttaacc aaaggattcc tgatttccac agttctcgtc atcagctctc tggttgcttt
     4741 agctaataca ccataagcat tttccctact gatgttcatc atctgagcgt attggttata
     4801 agtgaacgat accgtccgtt ctttccttgt agggttttca atcgtggggt tgagtagtgc
     4861 cacacagcat aaaattagct tggtttcatg ctccgttaag tcatagcgac taatcgctag
     4921 ttcatttgct ttgaaaacaa ctaattcaga catacatctc aattggtcta ggtgatttta
     4981 atcactatac caattgagat gggctagtca atgataatta ctagtccttt tcccgggtga
     5041 tctgggtatc tgtaaattct gctagacctt tgctggaaaa cttgtaaatt ctgctagacc
     5101 ctctgtaaat tccgctagac ctttgtgtgt tttttttgtt tatattcaag tggttataat
     5161 ttatagaata aagaaagaat aaaaaaagat aaaaagaata gatcccagcc ctgtgtataa
     5221 ctcactactt tagtcagttc cgcagtatta caaaaggatg tcgcaaacgc tgtttgctcc
     5281 tctacaaaac agaccttaaa accctaaagg cttaagtagc accctcgcaa gctcgggcaa
     5341 atcgctgaat attccttttg tctccgacca tcaggcacct gagtcgctgt ctttttcgtg
     5401 acattcagtt cgctgcgctc acggctctgg cagtgaatgg gggtaaatgg cactacaggc
     5461 gccttttatg gattcatgca aggaaactac ccataataca agaaaagccc gtcacgggct
     5521 tctcagggcg ttttatggcg ggt
//
