LOCUS       sequence-355953-        5339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses c-di-GMP 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-MrkH
FEATURES             Location/Qualifiers
     source          1..5339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     589..608
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     841..859
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1779..1807)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3086..3106
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3187..3233
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3406..4356)
                     /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      4404..4626
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5132..5166)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(join(5207..5339,1..662))
                     /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"
     primer_bind     complement(5318..5337)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
       61 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
      121 ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
      181 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
      241 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
      301 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
      361 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
      421 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc
      481 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
      541 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
      601 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
      661 atgcgaaacg atcctcatcc tgtctcttga tcagatcttg atcccctgcg ccatcagatc
      721 cttggcggca agaaagccat ccagtttact ttgcagggct tcccaacctt accagagggc
      781 gccccagctg gcaattccga cgtctaagaa accattatta tcatgacatt aacctataaa
      841 aataggcgta tcacgaggcc ctttcgtctt caccacagct aacaccacgt cgtccctatc
      901 tgctgcccta ggtctatgag tggttgctgg ataactttac gggcatgcat aaggctcgta
      961 tgatatattc agggagacca caacggtttc cctctacaaa taattttgtt taacttttct
     1021 agatttaaga aggagatata catatgagta aaggagaagc tgtgattaaa gagttcatgc
     1081 gcttcaaagt tcacatggag ggttctatga acggtcacga gttcgagatc gaaggcgaag
     1141 gcgagggccg tccgtatgaa ggcacccaga ccgccaaact gaaagtgact aaaggcggcc
     1201 cgctgccttt ttcctgggac atcctgagcc cgcaatttat gtacggttct agggcgttca
     1261 tcaaacaccc agcggatatc ccggactatt ataagcagtc ttttccggaa ggtttcaagt
     1321 gggaacgcgt aatgaatttt gaagatggtg gtgccgtgac cgtcactcag gacacctccc
     1381 tggaggatgg caccctgatc tataaagtta aactgcgtgg tactaatttt ccacctgatg
     1441 gcccggtgat gcagaaaaag acgatgggtt gggaggcgtc taccgaacgc ttgtatccgg
     1501 aagatggtgt gctgaaaggc gacattaaaa tggccctgcg cctgaaagat ggcggccgct
     1561 atctggctga cttcaaaacc acgtacaaag ccaagaaacc tgtgcagatg cctggcgcgt
     1621 acaatgtgga ccgcaaactg gacatcacct ctcataatga agattatacg gtggtagagc
     1681 aatatgagcg ctccgagggt cgtcattcta ccggtggcat ggatgaacta tacaaataag
     1741 gaacagacat gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat
     1801 tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa
     1861 catacggaaa acttaccctt aaattgattt gcactactgg aaaactacct gttccatggc
     1921 caacacttgt cactactttg ggttatggtg ttcaatgctt tgcgagatac ccagatcata
     1981 tgaaacagca tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta
     2041 tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt gaaggtgata
     2101 cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga aacattcttg
     2161 gacacaaatt ggaatacaac tatggtagtg gtaccgaggg taccattaag accagcaaat
     2221 acgaaatcat tgcgatcttc cgtgaggaac tgcgtaagcg taccgagatc gaaattttct
     2281 ttaacaacac cagcatcatc acccagctga cccgtgtgga tttcgcggag tttcacatcc
     2341 aaacccaccg taagattccg agcggtcaca aaatccgttt cctgctgcac agcgacagcg
     2401 gcaaaattga gtttaacgcg gcgctgacca agcacgataa cagcggtgtt gacaaaggca
     2461 tccgttacgc gtttagcctg ccggaatgcc tgcaggtggt tcaacgtcgt cgtgatccgc
     2521 gtttccgtct gcgtcacgag cacgactttt actgccgtgg tcgtcacaag aacggcgaaa
     2581 actacctgtt cgacatcaaa gatattagcg acggtggctg cgcgctgatg accaagaccc
     2641 cgaacctgaa atttctgagc cacaacgcgc tgctgaagaa cgcggtgctg atgctggcgg
     2701 agtatggcga aatcaccatt gatctggtgg ttaaaaacgt gatcgttatt accctggaca
     2761 acgcgaacga ggaaagcgaa agctactatc agatcagctg ccaattcaag tttcgtcacc
     2821 tggacgatca gcgtcgtatc gagaagattc tgctggatct gattctgaac tcacacaatg
     2881 tatacatcac ggcagacaaa caaaagaatg gaatcaaagc gaacttcaaa attagacaca
     2941 acattgaaga tggaggtgtt caactagcag accattatca acaaaatact ccaattggcg
     3001 atggccctgt ccttttacca gacaaccatt acctgtccta ccaatctaag ctttcgaaag
     3061 atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta
     3121 cacatggcat ggatgaacta tacaaataaa tgtccagacc tgcaggcatg caagctctag
     3181 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttga
     3241 aatgcaccaa aaactcgtaa aagctctgat gtatctatct tttttacacc gttttcatct
     3301 gtgcatatgg acagttttcc ctttgatatc taacggtgaa cagttgttct acttttgttt
     3361 gttagtcttg atgcttcact gatagataca agagccataa gaacctcaga tccttccgta
     3421 tttagccagt atgttctcta gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc
     3481 attgagatca tgcttacttt gcatgtcact caaaaatttt gcctcaaaac tggtgagctg
     3541 aatttttgca gttaaagcat cgtgtagtgt ttttcttagt ccgttacgta ggtaggaatc
     3601 tgatgtaatg gttgttggta ttttgtcacc attcattttt atctggttgt tctcaagttc
     3661 ggttacgaga tccatttgtc tatctagttc aacttggaaa atcaacgtat cagtcgggcg
     3721 gcctcgctta tcaaccacca atttcatatt gctgtaagtg tttaaatctt tacttattgg
     3781 tttcaaaacc cattggttaa gccttttaaa ctcatggtag ttattttcaa gcattaacat
     3841 gaacttaaat tcatcaaggc taatctctat atttgccttg tgagttttct tttgtgttag
     3901 ttcttttaat aaccactcat aaatcctcat agagtatttg ttttcaaaag acttaacatg
     3961 ttccagatta tattttatga atttttttaa ctggaaaaga taaggcaata tctcttcact
     4021 aaaaactaat tctaattttt cgcttgagaa cttggcatag tttgtccact ggaaaatctc
     4081 aaagccttta accaaaggat tcctgatttc cacagttctc gtcatcagct ctctggttgc
     4141 tttagctaat acaccataag cattttccct actgatgttc atcatctgag cgtattggtt
     4201 ataagtgaac gataccgtcc gttctttcct tgtagggttt tcaatcgtgg ggttgagtag
     4261 tgccacacag cataaaatta gcttggtttc atgctccgtt aagtcatagc gactaatcgc
     4321 tagttcattt gctttgaaaa caactaattc agacatacat ctcaattggt ctaggtgatt
     4381 ttaatcacta taccaattga gatgggctag tcaatgataa ttactagtcc ttttcctttg
     4441 agttgtgggt atctgtaaat tctgctagac ctttgctgga aaacttgtaa attctgctag
     4501 accctctgta aattccgcta gacctttgtg tgtttttttt gtttatattc aagtggttat
     4561 aatttataga ataaagaaag aataaaaaaa gataaaaaga atagatccca gccctgtgta
     4621 taactcacta ctttagtcag ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc
     4681 tcctctacaa aacagacctt aaaaccctaa aggcttaagt agcaccctcg caagctcggg
     4741 caaatcgctg aatattcctt ttgtctccga ccatcaggca cctgagtcgc tgtctttttc
     4801 gtgacattca gttcgctgcg ctcacggctc tggcagtgaa tgggggtaaa tggcactaca
     4861 ggcgcctttt atggattcat gcaaggaaac tacccataat acaagaaaag cccgtcacgg
     4921 gcttctcagg gcgttttatg gcgggtctgc tatgtggtgc tatctgactt tttgctgttc
     4981 agcagttcct gccctctgat tttccagtct gaccacttcg gattatcccg tgacaggtca
     5041 ttcagactgg ctaatgcacc cagtaaggca gcggtatcat caacaggctt acccgtctta
     5101 ctgtcaacca gacccgccag gattctcatg cataaaaaac gcccggcggc aaccgagcgt
     5161 tctgaattaa ttaatcatcg cgactgcaga gatcccagag tccttattag aagaactcgt
     5221 caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga
     5281 ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgct
//
