LOCUS       sequence-146031-        3574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of double-cysteine variant of plasmid segregation 
            protein, ParM, as scaffold for ADP biosensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3574)
  AUTHORS   Kunzelmann S, Webb MR
  TITLE     A fluorescent, reagentless biosensor for ADP based on 
            tetramethylrhodamine-labeled ParM.
  JOURNAL   ACS Chem Biol. 2010 Apr 16;5(4):415-25. doi: 10.1021/cb9003173.
  PUBMED    20158267
REFERENCE   2  (bases 1 to 3574)
  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"; journalName: 
            "ACS Chem Biol"; date: "2010-04-16- 16"; volume: "5"; issue: "4"; 
            pages: "415-25"
COMMENT     Sequence Label: pHis17_ParM _2
FEATURES             Location/Qualifiers
     source          1..3574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(14..32)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        100..204
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(205..273,274..1065)
                     /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     complement(423..442)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1236..1824
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1725..1744
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1978..1995
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2070..2089)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2230..2252
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2305..2324
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2305..2323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2355..2377
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3357..3374
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3384..3401
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3411..3428
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3482..3500)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3496..3543
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
       61 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
      121 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
      181 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
      241 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
      301 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
      361 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
      421 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
      481 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
      541 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
      601 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
      661 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
      721 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
      781 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
      841 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
      901 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
      961 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     1021 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     1081 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1141 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     1201 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1261 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1321 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1381 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1441 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1501 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     1561 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     1621 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     1681 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     1741 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     1801 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     1861 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     1921 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     1981 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     2041 tgcggtattt cacaccggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     2101 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     2161 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     2221 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     2281 ggcaggatct cgatcccgcg aaattaatac gactcactat agggagacca caacggtttc
     2341 cctctagaaa taattttgtt taactttaag aaggagatat acatatgttg gtattcattg
     2401 atgacggttc aacaaacatc aaactacagt ggcaggaaag cgacggaaca attaaacagc
     2461 acattagccc gaacagcttc gcacgcgagt gggcagtctc ttttggtgat aaaaaggtct
     2521 ttaactacac actgaacggc gaacagtatt catttgatcc aatcagcccg tgtgctgtag
     2581 tcacaactaa tatcgcatgg caatacagcg acgttaatgt cgttgcagtg catcacgcct
     2641 tactgaccag tggtctgccg gtaagcgaag tggatattgt ttgcacactt cctctgacag
     2701 agtattacga cagaaataac caacccaata cggaaaatat tgagcgtaag aaagcaaact
     2761 tccggaaaaa aattacatta aatggcgggg atacattcac aataaaagat gtaaaagtca
     2821 tgcctgaatc tataccggca ggttatgaag ttctacaaga actggatgag ttagattctt
     2881 tattaattat agatctcggg ggcgccaact tagatatttc tcaggtaatg gggaaattat
     2941 cggggatcag taaaatatac ggagactcat ctcttggtgt ctctctggtt acatctgcag
     3001 taaaagatgc cctttctctt gcgagaacaa aaggaagtag ctatcttgct gactgtataa
     3061 tcattcacag aaaagataat aactatctga agcaacgaat taatgatgag aacaaaatat
     3121 caatagtcac cgaagcaatg aatgaagcac ttcgtaaact tgagcaacgt gtattaaata
     3181 cgctcaatga attttctggt tatactcatg ttatggttat aggcggtggc gcagaattaa
     3241 tagccgatgc agtaaaaaaa cacacacaga ttcgtgatga acgttttttc aaaaccaata
     3301 actctcaata tgatttagtt aacggtatgt atctcatagg taatcaatca ggatcccatc
     3361 atcatcatca tcattaaaag cttcaccacc accaccacca ctaagaattc catcatcatc
     3421 atcatcatta acgatccggc tgctaacaaa gcccgaaagg aagctaggtt ggctgctgcc
     3481 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     3541 ttgctgaaag gaggaactat atccggataa ttct
//
