LOCUS       sequence-252582-        6061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Salmonella typhimurium diguanylate cyclase AdrA cloned as AdrA-8xHis
            in pBAD30.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6061)
  AUTHORS   Simm R, Morr M, Kader A, Nimtz M, Romling U
  TITLE     GGDEF and EAL domains inversely regulate cyclic di-GMP levels and 
            transition from sessility to motility.
  JOURNAL   Mol Microbiol. 2004 Aug;53(4):1123-34. doi: 
            10.1111/j.1365-2958.2004.04206.x.
  PUBMED    15306016
REFERENCE   2  (bases 1 to 6061)
  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: 
            "10.1111/j.1365-2958.2004.04206"; journalName: "Mol Microbiol"; 
            date: "2004-08"; volume: "53"; issue: "4"; pages: "1123-34"
COMMENT     Sequence Label: pWJB30
FEATURES             Location/Qualifiers
     source          1..6061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(59..145)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     298..315
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(356..379)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     RBS             1496..1504
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(1558..1842)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(1616..1635)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1869..2747
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     complement(2927..2946)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      3022..3567
                     /direction=RIGHT
                     /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     3468..3487
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3684..3701
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4533..4988)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4670..4691)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4882..4901
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(join(5030..5821,5822..5890))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5653..5672
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5891..5982)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(6001..6028)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gtcctgcccg ccaccctccg ggccgttgct tcgcaacgtt caaatccgct cccggcggat
       61 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
      121 tcgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc atggggagac
      181 cccacactac catcggcgct acggcgtttc acttctgagt tcggcatggg gtcaggtggg
      241 accaccgcgc tactgccgcc aggcaaattc tgttttatca gaccgcttct gcgttctgat
      301 ttaatctgta tcaggctgaa aatcttctct catccgccaa aacagccaag cttcaatgat
      361 gatgatgatg atgatgatgt gccgccactt cggtgcggtt acgtccggca ttctttgctt
      421 tgtaaagcgc catatccgca gactttagcc attcccgata atgaccgatt tgtggcgtta
      481 acggcgccac gcccacgctg atacgtaaca tgacctgcgg cgcgcccggc aagcgtaagg
      541 tatttagccg ttcatgcact cgcgacatgg cggtaatcgc gctgtctgcc ggcgttccgc
      601 acataatcac cgcaaattcg tcgccgccaa agcgcccgat aatatcactc ccgcgtagcg
      661 ttatctgtaa ttgacgcgtc agggcgataa tagcctcatc cccaacatca tgtccccagg
      721 tgtcgttaat gctcttaaaa tgatcgatat cgataatcaa caatgtcgcc tcccgatggt
      781 ggcgtcggct gtgatcgaat tcattacgta ataaaatttc ccagtggcgt cggttataga
      841 cgccggtcat gccatcccgc gagctcatcg cctgcaagcg tcgcttatgt tccgctaatt
      901 taatggccgt ttggtaactt acccaggcga acagaagcgg atacaacata atgacgggca
      961 aagagagcgt gacttccagc gacgagcttc gcatcgcgac gggtagcccg gcaagctgca
     1021 acgtcaccag acatgacgcc agcaaaaggc ccattccgac agtgaatagc cgccgcccgc
     1081 cagcgcccat cagattcatg ctcatcatca taaacagcgc ggcggcaggc aacatattga
     1141 cgcccatcag cgcgatccac atgccggaga ggatcgcatc cactttcagg ttgtaaattt
     1201 cctggcgtag cggatcaagg gctttcgccg cccactgcca tgccagatgc ggccagacaa
     1261 aagaccagcc gaccagcaat agccaccagc cgccaaagac aggttgcgtc actaacacgg
     1321 cggccaccgg taaaaacatc ccccctaacc ccaccgcacg cggtagcctg atgcgccggg
     1381 cgaagcgcaa cccggagcgc tgacgatcat caggagggtc aggcggcgct accgcctgtt
     1441 caaccgcttt tcggtaaaaa ttttcatcat tcattatttt tgggaacatt tagaaacctc
     1501 cttatctaga ggatccccgg gtaccgagct cgaattcgct agcccaaaaa aacgggtatg
     1561 gagaaacagt agagagttgc gataaaaagc gtcaggtagg atccgctaat cttatggata
     1621 aaaatgctat ggcatagcaa agtgtgacgc cgtgcaaata atcaatgtgg acttttctgc
     1681 cgtgattata gacacttttg ttacgcgttt ttgtcatggc tttggtcccg ctttgttaca
     1741 gaatgctttt aataagcggg gttaccggtt tggttagcga gaagagccag taaaagacgc
     1801 agtgacggca atgtctgatg caatatggac aattggtttc ttctctgaat ggcgggagta
     1861 tgaaaagtat ggctgaagcg caaaatgatc ccctgctgcc gggatactcg tttaatgccc
     1921 atctggtggc gggtttaacg ccgattgagg ccaacggtta tctcgatttt tttatcgacc
     1981 gaccgctggg aatgaaaggt tatattctca atctcaccat tcgcggtcag ggggtggtga
     2041 aaaatcaggg acgagaattt gtttgccgac cgggtgatat tttgctgttc ccgccaggag
     2101 agattcatca ctacggtcgt catccggagg ctcgcgaatg gtatcaccag tgggtttact
     2161 ttcgtccgcg cgcctactgg catgaatggc ttaactggcc gtcaatattt gccaatacgg
     2221 ggttctttcg cccggatgaa gcgcaccagc cgcatttcag cgacctgttt gggcaaatca
     2281 ttaacgccgg gcaaggggaa gggcgctatt cggagctgct ggcgataaat ctgcttgagc
     2341 aattgttact gcggcgcatg gaagcgatta acgagtcgct ccatccaccg atggataatc
     2401 gggtacgcga ggcttgtcag tacatcagcg atcacctggc agacagcaat tttgatatcg
     2461 ccagcgtcgc acagcatgtt tgcttgtcgc cgtcgcgtct gtcacatctt ttccgccagc
     2521 agttagggat tagcgtctta agctggcgcg aggaccaacg tatcagccag gcgaagctgc
     2581 ttttgagcac cacccggatg cctatcgcca ccgtcggtcg caatgttggt tttgacgatc
     2641 aactctattt ctcgcgggta tttaaaaaat gcaccggggc cagcccgagc gagttccgtg
     2701 ccggttgtga agaaaaagtg aatgatgtag ccgtcaagtt gtcataattg gtaacgaatc
     2761 agacaattga cggcttgacg gagtagcata gggtttgcag aatccctgct tcgtccattt
     2821 gacaggcaca ttatgcatcg atgataagct gtcaaacatg agcagatcct ctacgccgga
     2881 cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac
     2941 atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgatcctaga aatattttat
     3001 ctgattaata agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga
     3061 aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt ctctgagcta ccaactcttt
     3121 gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc
     3181 ttaaccggcg catgacttca agactaactc ctctaaatca attaccagtg gctgctgcca
     3241 gtggtgcttt tgcatgtctt tccgggttgg actcaagacg atagttaccg gataaggcgc
     3301 agcggtcgga ctgaacgggg ggttcgtgca tacagtccag cttggagcga actgcctacc
     3361 cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg aatgacaccg
     3421 gtaaaccgaa aggcaggaac aggagagcgc acgagggagc cgccaggggg aaacgcctgg
     3481 tatctttata gtcctgtcgg gtttcgccac cactgatttg agcgtcagat ttcgtgatgc
     3541 ttgtcagggg ggcggagcct atggaaaaac ggctttgccg cggccctctc acttccctgt
     3601 taagtatctt cctggcatct tccaggaaat ctccgccccg ttcgtaagcc atttccgctc
     3661 gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa tatatcctgt
     3721 atcacatatt ctgctgacgc accggtgcag ccttttttct cctgccacat gaagcacttc
     3781 actgacaccc tcatcagtgc caacatagta agccagtata cactccgcta gccatgggca
     3841 aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg
     3901 tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt
     3961 ggcagggcgg ggcgtaattt ttttaaggca gttattggtg cccttaaacg cctggttgct
     4021 acgcctgaat aagtgataat aagcggatga atggcagaaa ttcgaaagca aattcgaccc
     4081 ggtcgtcggt tcagggcagg gtcgttaaat agccgcttat gtctattgct ggtttaccgg
     4141 tttattgact accggaagca gtgtgaccgt gtgcttctca aatgcctgac tggtaacagg
     4201 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4261 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4321 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4381 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4441 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4501 ttatcaaaaa ggatcttcac ctagatcctt ttaaattgta aacgttaata ttttgttaaa
     4561 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     4621 aatcccttat aaatcaaaag aatagcccga gatagggttg agtgttgttc cagtttggaa
     4681 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     4741 gggcgatggc ccactacgtg aaccatcacc caaatcaagt tttttggggt cgaggtgccg
     4801 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     4861 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     4921 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     4981 gggcgcgtaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5041 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5101 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5161 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5221 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5281 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5341 ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5401 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5461 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5521 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5581 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5641 cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5701 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5761 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5821 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5881 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5941 tgagcggata catatttgaa tgtatttaga aaaataaaca aaagagtttg tagaaacgca
     6001 aaaaggccat ccgtcaggat ggccttctgc ttaatttgat gcctggcagt ttatggcggg
     6061 c
//
