LOCUS       sequence-196978-        7516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses E.coli relAE335Q from the NS1 site in the S. elongatus 
            PCC7942 genome.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7516)
  AUTHORS   Puszynska AM, O'Shea EK
  TITLE     ppGpp Controls Global Gene Expression in Light and in Darkness in S.
            elongatus.
  JOURNAL   Cell Rep. 2017 Dec 12;21(11):3155-3165. doi: 
            10.1016/j.celrep.2017.11.067.
  PUBMED    29241543
REFERENCE   2  (bases 1 to 7516)
  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.celrep.2017.11"; journalName: "Cell Rep"; date: 
            "2017-12-12- 12"; volume: "21"; issue: "11"; pages: "3155-3165"
COMMENT     Sequence Label: relAE335Q_NS1
FEATURES             Location/Qualifiers
     source          1..7516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     86..108
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        join(1243..1248,1249..1265,1266..1272)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1280..1296
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             2755..2778
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3657..4448
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      6602..7190
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7091..7110
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7344..7361
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7376..7516
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7442..7461)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
       61 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
      121 tcaccgaaac gcgcgaggca ggatcgatcc ggcagccggc ggagcgctgc tttcttggca
      181 agcggtcgcc agccccaacg ccagggctgc cagcccgaaa cagcggggca aggcagcttg
      241 gaagggcgat cgcagcacgg gcatggcaat gtctctctga aggaatgcag accttattcg
      301 tacagccagg gttgaatcgt gggggtccaa tcacttagct ctgctgggct aaaccagaga
      361 gcaatttcct gttgtgctgt ttcgattgca tccgagccat ggatgatgtt gcggccaata
      421 ttgacaccaa aatcaccacg gatggtgccc ggttctgccg tcagcggatt ggtagcgccg
      481 atcaacttgc gagcagccgc cacaacgcct tcgccttcca agacgatcgc cacgatcggc
      541 ccagaggtga tgaactcgac gaggccattg aagaaggggc gctcgcggtg gacagcatag
      601 tgctgttcgg ccagctcgcg actgggcttc agctgcttta ggcccaccag tttgaagcct
      661 ttttgctcaa agcggccgat gatcgtaccg accaaacccc gctgaacgcc atcgggcttg
      721 atggcaataa atgtgcgttc cacagacatc tagatagtcc tcaagacgag gcaagcattg
      781 agcttgcctt cctatggttc gggatcactg ggattcttga caagcgatcg cggtcacatc
      841 gctatctctt aggacttcgc agcgggcgag tcggattgac ccggtaggga tttcgccaga
      901 tcaatgcccg tggtttgttt cagcttctcc agcaagctag cgatttgggt agcgctgcct
      961 tccccttcgc caatcacagt gatcgactcc acgtcgatat ctggcacggt gcctgaaagc
     1021 gtgacgagca gggactcgac ctgctccaga agctcgaaat tcgagctcga ctgcacggtg
     1081 caccaatgct tctggcgtca ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta
     1141 aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat gttttttgcg
     1201 ccgacatcat aacggttctg gcaaatattc tgaaatgagc tgttgacaat taatcatccg
     1261 gctcgtataa tgtgtggaat tgtgagcgga taacaatttc atacgctcac aattggtacc
     1321 ggtgatacca gcatcgtctt gatgcccttg gcagcaccct gctaaggagg caacaagatg
     1381 gttgcggtaa gaagtgcaca tatcaataag gctggtgaat ttgatccgga aaaatggatc
     1441 gcaagtctgg gtattaccag ccagaagtcg tgtgagtgct tagccgaaac ctgggcgtat
     1501 tgtctgcaac agacgcaggg gcatccggat gccagtctgt tattgtggcg tggtgttgag
     1561 atggtggaga tcctctcgac attaagtatg gacattgaca cgctgcgggc ggcgctgctt
     1621 ttccctctgg cggatgccaa cgtagtcagc gaagatgtgc tgcgtgagag cgtcggtaag
     1681 tcggtcgtta accttattca cggcgtgcgt gatatggcgg cgatccgcca gctgaaagcg
     1741 acgcacactg attctgtttc ctccgaacag gtcgataacg ttcgccggat gttattggcg
     1801 atggtcgatg attttcgctg cgtagtcatc aaactggcgg agcgtattgc tcatctgcgc
     1861 gaagtaaaag atgcgccgga agatgaacgt gtactggcgg caaaagagtg taccaacatc
     1921 tacgcaccgc tggctaaccg tctcggaatc ggacaactga aatgggaact ggaagattac
     1981 tgcttccgtt acctccatcc aaccgaatac aaacgaattg ccaaactgct gcatgaacgg
     2041 cgtctcgacc gcgaacacta catcgaagag ttcgttggtc atctgcgcgc tgagatgaaa
     2101 gctgaaggcg ttaaagcgga agtgtatggt cgtccgaaac acatctacag catctggcgt
     2161 aaaatgcaga aaaagaacct cgcctttgat gagctgtttg atgtgcgtgc ggtacgtatt
     2221 gtcgccgagc gtttacagga ttgctatgcc gcactgggga tagtgcacac tcactatcgc
     2281 cacctgccgg atgagtttga cgattacgtc gctaacccga aaccaaacgg ttatcagtct
     2341 attcataccg tggttctggg gccgggtgga aaaaccgttc agatccaaat ccgcaccaaa
     2401 cagatgcatg aagatgcaga gttgggtgtt gctgcgcact ggaaatataa agagggcgcg
     2461 gctgctggcg gcgcacgttc gggacatgaa gaccggattg cctggctgcg taaactgatt
     2521 gcgtggcagg aagagatggc tgattccggc gaaatgctcg acgaagtacg tagtcaggtc
     2581 tttgacgacc gggtgtacgt ctttacgccg aaaggtgatg tcgttgattt gcctgcggga
     2641 tcaacgccgc tggacttcgc ttaccacatc cacagtgatg tcggacaccg ctgcatcggg
     2701 gcaaaaattg gcgggcgcat tgtgccgttc acctaccagc tgggcagcgg cagcgattac
     2761 aaagatgatg atgataaata gtgcatgcct gcaggtcgac tctagagctt tatgcttgta
     2821 aaccgttttg tgaaaaaatt tttaaaataa aaaaggggac ctctagggtc cccaattaat
     2881 tagtaatata atctattaaa ggtcattcaa aaggtcatcc accggatcaa ttcccctgct
     2941 cgcgcaggct gggtgccaag ctctcgggta acatcaaggc ccgatccttg gagcccttgc
     3001 cctcccgcac gatgatcgtg ccgtgatcga aatccagatc cttgacccgc agttgcaaac
     3061 cctcactgat ccgcatgccc gttccataca gaagctgggc gaacaaacga tgctcgcctt
     3121 ccagaaaacc gaggatgcga accacttcat ccggggtcag caccaccggc aagcgccgcg
     3181 acggccgagg tcttccgatc tcctgaagcc agggcagatc cgtgcacagc accttgccgt
     3241 agaagaacag caaggccgcc aatgcctgac gatgcgtgga gaccgaaacc ttgcgctcgt
     3301 tcgccagcca ggacagaaat gcctcgactt cgctgctgcc caaggttgcc gggtgacgca
     3361 caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg gttcgtaagc
     3421 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     3481 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt ggggtacagt
     3541 ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat
     3601 ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta aacatcatga
     3661 gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc
     3721 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc
     3781 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa
     3841 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga
     3901 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc
     3961 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct
     4021 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata
     4081 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc
     4141 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg
     4201 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa
     4261 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc
     4321 ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc
     4381 gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg
     4441 gcaaataatg tctaacaatt cgttcaagcc gacgccgctt cgcggcgcgg cttaactcaa
     4501 gcgttagatg cactaagcac ataattgctc acagccaaac tatcaggtca agtctgcttt
     4561 tattattttt aagcgtgcat aataagccct acacaaattg ggagatatat catgaaaggc
     4621 tggctttttc ttgttatcgc aatagttggc gaagtaatcg caacatccgc attaaaatct
     4681 agcgagggct ttactaagct gatccggtgg atgacctttt gaatgacctt taatagatta
     4741 tattactaat taattgggga ccctagaggt cccctttttt attttaaaaa ttttttcaca
     4801 aaacggttta caagcatact agaggatcgg cggccgcgga tccccgggta ccgagctcga
     4861 atttcgagct tctggagcag gaagatgtcg cgggcattag caccagcggt ctgccaagcc
     4921 tccgccagcc gttgggtccc ttccgcttga gcttttccat cttcgacgat acgggcggcg
     4981 gccccccgcg cttccgcgat cgcccgttta caagctgcct cagctggggc gatcacatcg
     5041 gcttgaagtt gctgctgcac ctgtttgatc cgctcctgct gcacagggag ttctgcttgg
     5101 ctacgagcga cttcggtagc aatgtccgct tcagcttcgg ccaccaccgc ttcgcgccgc
     5161 gtcaacgcat cctgaatccg gcgctcggcc tcggcttggg cgatcgctac atcgcgatcg
     5221 atccgacgca gggccgtgat cttgtcattt tcggccgttt ggatcgcaga ggcagcctgg
     5281 gcatcggctt cagcaattcg ggcatctcgc tgcagatcag cccgctgctt gcgtccacta
     5341 gccgagagat aaccgacctc atcggaaatg ttctggactt gcagcgtatc gaggactaga
     5401 cccagctgct caaggtcatc ctccgcctct tccagcagac ttttggcaaa ggcaattttg
     5461 tcctcgttga tctgctccgg cgtgaggctg gctaaaacac cacgcaagtt gccttcgagg
     5521 gtctccttgg caatttgctc gatttcctta cggtttttgc caagcagccg ctcgatcgcg
     5581 ttgtggatgg tcggttcttc cccagcaatc ttgatattgg caacgccttc aacagtcagg
     5641 ggaatgccgc ccttggagaa ggcattggaa acgcgcaact caatgatcat gttggtcaga
     5701 tccatgcgga gcgctttttc cagcagaggt acccgcaggc tgctgccgcc cttgaccaag
     5761 cgatagccaa ctcggcggcc atcactactg cggcgactac tgccagcaaa gatcaaaatt
     5821 tcactgggtt ggcagatgta gtagagattg cgcaggacta agctgccagc cccggcgact
     5881 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5941 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     6001 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6061 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     6121 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6181 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6241 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6301 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6361 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6421 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6481 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6541 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6601 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6661 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6721 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     6781 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6841 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6901 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6961 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7021 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7081 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7141 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7201 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     7261 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     7321 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     7381 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
     7441 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
     7501 ctgggtcatg gctgcg
//
