LOCUS       sequence-355954-        5339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses c-di-GMP mutant biosensor in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5339)
  AUTHORS   Vrabioiu AM, Berg HC
  TITLE     Signaling events that occur when cells of Escherichia coli encounter
            a glass surface.
  JOURNAL   Proc Natl Acad Sci U S A. 2022 Feb 8;119(6). pii: 2116830119. doi: 
            10.1073/pnas.2116830119.
  PUBMED    35131853
REFERENCE   2  (bases 1 to 5339)
  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"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2022-02-8"; volume: "119(6). pii: 
            2116830119. doi"; pages: " 10.1073/pnas.2116830119"
COMMENT     Sequence Label: pEB2_mScarletI_mVenusNB-MrkHR113A
FEATURES             Location/Qualifiers
     source          1..5339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(78..97)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     688..707
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     940..958
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1878..1906)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3185..3205
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3286..3332
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3505..4455)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      4503..4725
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5231..5265)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(join(5306..5339,1..761))
                     /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"
ORIGIN
        1 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
       61 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc
      121 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
      181 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg
      241 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
      301 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
      361 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
      421 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
      481 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
      541 cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg
      601 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
      661 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
      721 gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct
      781 gtctcttgat cagatcttga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc
      841 cagtttactt tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccgac
      901 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
      961 tttcgtcttc accacagcta acaccacgtc gtccctatct gctgccctag gtctatgagt
     1021 ggttgctgga taactttacg ggcatgcata aggctcgtat gatatattca gggagaccac
     1081 aacggtttcc ctctacaaat aattttgttt aacttttcta gatttaagaa ggagatatac
     1141 atatgagtaa aggagaagct gtgattaaag agttcatgcg cttcaaagtt cacatggagg
     1201 gttctatgaa cggtcacgag ttcgagatcg aaggcgaagg cgagggccgt ccgtatgaag
     1261 gcacccagac cgccaaactg aaagtgacta aaggcggccc gctgcctttt tcctgggaca
     1321 tcctgagccc gcaatttatg tacggttcta gggcgttcat caaacaccca gcggatatcc
     1381 cggactatta taagcagtct tttccggaag gtttcaagtg ggaacgcgta atgaattttg
     1441 aagatggtgg tgccgtgacc gtcactcagg acacctccct ggaggatggc accctgatct
     1501 ataaagttaa actgcgtggt actaattttc cacctgatgg cccggtgatg cagaaaaaga
     1561 cgatgggttg ggaggcgtct accgaacgct tgtatccgga agatggtgtg ctgaaaggcg
     1621 acattaaaat ggccctgcgc ctgaaagatg gcggccgcta tctggctgac ttcaaaacca
     1681 cgtacaaagc caagaaacct gtgcagatgc ctggcgcgta caatgtggac cgcaaactgg
     1741 acatcacctc tcataatgaa gattatacgg tggtagagca atatgagcgc tccgagggtc
     1801 gtcattctac cggtggcatg gatgaactat acaaataagg aacagacatg agtaaaggag
     1861 aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc
     1921 acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta
     1981 aattgatttg cactactgga aaactacctg ttccatggcc aacacttgtc actactttgg
     2041 gttatggtgt tcaatgcttt gcgagatacc cagatcatat gaaacagcat gactttttca
     2101 agagtgccat gcccgaaggt tatgtacagg aaagaactat atttttcaaa gatgacggga
     2161 actacaagac acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat agaatcgagt
     2221 taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg gaatacaact
     2281 atggtagtgg taccgagggt accattaaga ccagcaaata cgaaatcatt gcgatcttcc
     2341 gtgaggaact gcgtaagcgt accgagatcg aaattttctt taacaacacc agcatcatca
     2401 cccagctgac ccgtgtggat ttcgcggagt ttcacatcca aacccaccgt aagattccga
     2461 gcggtcacaa aatccgtttc ctgctgcaca gcgacagcgg caaaattgag tttaacgcgg
     2521 cgctgaccaa gcacgataac agcggtgttg acaaaggcat ccgttacgcg tttagcctgc
     2581 cggaatgcct gcaggtggtt caacgtcgtc gtgatccggc attccgtctg cgtcacgagc
     2641 acgactttta ctgccgtggt cgtcacaaga acggcgaaaa ctacctgttc gacatcaaag
     2701 atattagcga cggtggctgc gcgctgatga ccaagacccc gaacctgaaa tttctgagcc
     2761 acaacgcgct gctgaagaac gcggtgctga tgctggcgga gtatggcgaa atcaccattg
     2821 atctggtggt taaaaacgtg atcgttatta ccctggacaa cgcgaacgag gaaagcgaaa
     2881 gctactatca gatcagctgc caattcaagt ttcgtcacct ggacgatcag cgtcgtatcg
     2941 agaagattct gctggatctg attctgaact cacacaatgt atacatcacg gcagacaaac
     3001 aaaagaatgg aatcaaagcg aacttcaaaa ttagacacaa cattgaagat ggaggtgttc
     3061 aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag
     3121 acaaccatta cctgtcctac caatctaagc tttcgaaaga tcccaacgaa aagagagacc
     3181 acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat
     3241 acaaataaat gtccagacct gcaggcatgc aagctctaga ggcatcaaat aaaacgaaag
     3301 gctcagtcga aagactgggc ctttcgtttt atctgttgaa atgcaccaaa aactcgtaaa
     3361 agctctgatg tatctatctt ttttacaccg ttttcatctg tgcatatgga cagttttccc
     3421 tttgatatct aacggtgaac agttgttcta cttttgtttg ttagtcttga tgcttcactg
     3481 atagatacaa gagccataag aacctcagat ccttccgtat ttagccagta tgttctctag
     3541 tgtggttcgt tgtttttgcg tgagccatga gaacgaacca ttgagatcat gcttactttg
     3601 catgtcactc aaaaattttg cctcaaaact ggtgagctga atttttgcag ttaaagcatc
     3661 gtgtagtgtt tttcttagtc cgttacgtag gtaggaatct gatgtaatgg ttgttggtat
     3721 tttgtcacca ttcattttta tctggttgtt ctcaagttcg gttacgagat ccatttgtct
     3781 atctagttca acttggaaaa tcaacgtatc agtcgggcgg cctcgcttat caaccaccaa
     3841 tttcatattg ctgtaagtgt ttaaatcttt acttattggt ttcaaaaccc attggttaag
     3901 ccttttaaac tcatggtagt tattttcaag cattaacatg aacttaaatt catcaaggct
     3961 aatctctata tttgccttgt gagttttctt ttgtgttagt tcttttaata accactcata
     4021 aatcctcata gagtatttgt tttcaaaaga cttaacatgt tccagattat attttatgaa
     4081 tttttttaac tggaaaagat aaggcaatat ctcttcacta aaaactaatt ctaatttttc
     4141 gcttgagaac ttggcatagt ttgtccactg gaaaatctca aagcctttaa ccaaaggatt
     4201 cctgatttcc acagttctcg tcatcagctc tctggttgct ttagctaata caccataagc
     4261 attttcccta ctgatgttca tcatctgagc gtattggtta taagtgaacg ataccgtccg
     4321 ttctttcctt gtagggtttt caatcgtggg gttgagtagt gccacacagc ataaaattag
     4381 cttggtttca tgctccgtta agtcatagcg actaatcgct agttcatttg ctttgaaaac
     4441 aactaattca gacatacatc tcaattggtc taggtgattt taatcactat accaattgag
     4501 atgggctagt caatgataat tactagtcct tttcctttga gttgtgggta tctgtaaatt
     4561 ctgctagacc tttgctggaa aacttgtaaa ttctgctaga ccctctgtaa attccgctag
     4621 acctttgtgt gttttttttg tttatattca agtggttata atttatagaa taaagaaaga
     4681 ataaaaaaag ataaaaagaa tagatcccag ccctgtgtat aactcactac tttagtcagt
     4741 tccgcagtat tacaaaagga tgtcgcaaac gctgtttgct cctctacaaa acagacctta
     4801 aaaccctaaa ggcttaagta gcaccctcgc aagctcgggc aaatcgctga atattccttt
     4861 tgtctccgac catcaggcac ctgagtcgct gtctttttcg tgacattcag ttcgctgcgc
     4921 tcacggctct ggcagtgaat gggggtaaat ggcactacag gcgcctttta tggattcatg
     4981 caaggaaact acccataata caagaaaagc ccgtcacggg cttctcaggg cgttttatgg
     5041 cgggtctgct atgtggtgct atctgacttt ttgctgttca gcagttcctg ccctctgatt
     5101 ttccagtctg accacttcgg attatcccgt gacaggtcat tcagactggc taatgcaccc
     5161 agtaaggcag cggtatcatc aacaggctta cccgtcttac tgtcaaccag acccgccagg
     5221 attctcatgc ataaaaaacg cccggcggca accgagcgtt ctgaattaat taatcatcgc
     5281 gactgcagag atcccagagt ccttattaga agaactcgtc aagaaggcga tagaaggcg
//
