LOCUS       sequence-189871-        3155 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutively expresses the voltage indicator PROPS under the Sigma
            70 promoter in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3155)
  AUTHORS   Bruni GN, Weekley RA, Dodd BJT, Kralj JM
  TITLE     Voltage-gated calcium flux mediates Escherichia coli 
            mechanosensation.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Aug 29;114(35):9445-9450. doi: 
            10.1073/pnas.1703084114. Epub 2017 Aug 14.
  PUBMED    28808010
REFERENCE   2  (bases 1 to 3155)
  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: 
            "10.1073/pnas.1703084114"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2017-08-29- 29"; volume: "114"; issue: "35"; pages: 
            "9445-9450"
COMMENT     Sequence Label: pKL003
FEATURES             Location/Qualifiers
     source          1..3155
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..42
                     /label=BBa_J23118
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.56"
     CDS             823..840
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(896..913)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     primer_bind     complement(1110..1127)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1281..1869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1361..1380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2040..2831,2832..2900))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2663..2682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2901..3005)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3073..3091
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3095..3138)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    join(3141..3155,1..7)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 tctagagttg acggctagct cagtcctagg tattgtgcta gcattaactt taagaaggag
       61 atatacatac ccatgggtaa attattactg atattaggta gtgttattgc acttcctaca
      121 tttgctgcag gtggtggtga ccttgatgct agtgattaca ctggtgtttc tttttggtta
      181 gttactgctg ctttattagc atctactgta tttttctttg ttgaaagaga tagagtttct
      241 gcaaaatgga aaacatcatt aactgtatct ggtcttgtta ctggtattgc tttctggcat
      301 tacatgtaca tgagaggggt atggattgaa actggtgatt cgccaactgt atttagatac
      361 attaattggt tactaacagt tcctctatta atatgtgaat tctacttaat tcttgctgct
      421 gcaactaatg ttgctggatc attatttaag aaattactag ttggttctct tgttatgctt
      481 gtgtttggtt acatgggtga agcaggaatc atggctgcat ggcctgcatt cattattggg
      541 tgtttagctt gggtatacat gatttatgaa ttatgggctg gagaaggaaa atctgcatgt
      601 aatactgcaa gtcctgctgt gcaatcagct tacaacacaa tgatgtatat tatcatcttt
      661 ggttgggcga tttatcctgt aggttatttc acaggttacc tgatgggtga cggtggatca
      721 gctcttaact taaaccttat ctataacctt gctgactttg ttaacaagat tctatttggt
      781 ttaattatat ggaatgttgc tgttaaagaa tcttctaatg ctcaccatca ccatcaccat
      841 tgagtttaaa cggtctccag cttggctgtt ttggcggatg agagaagatt ttcagcctga
      901 tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta
      961 gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg
     1021 gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag
     1081 gctcagtcga aagactgggc cttggcttcc tcgctcactg actcgctgcg ctcggtcgtt
     1141 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     1201 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1261 aaggccgcgt tgctggcgtt tttccacagg ctccgccccc ctgacgagca tcacaaaaat
     1321 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1381 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1441 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1501 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1561 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1621 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1681 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     1741 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1801 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1861 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1921 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     1981 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     2041 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2101 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     2161 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     2221 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     2281 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     2341 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     2401 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     2461 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     2521 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     2581 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     2641 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     2701 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     2761 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     2821 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     2881 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     2941 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     3001 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     3061 ttaacctata aaaataggcg tatcacgagg cagaatttca gataaaaaaa atccttagct
     3121 ttcgctaagg atgatttctg gaattcgcgg ccgct
//
