LOCUS       sequence-339477-        4458 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ionic strength FRET sensor with mCerulean3 and mCitrine for E. coli 
            expression. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4458)
  AUTHORS   Liu B, Poolman B, Boersma AJ
  TITLE     Ionic Strength Sensing in Living Cells.
  JOURNAL   ACS Chem Biol. 2017 Oct 20;12(10):2510-2514. doi: 
            10.1021/acschembio.7b00348. Epub 2017 Sep 6.
  PUBMED    28853549
REFERENCE   2  (bases 1 to 4458)
  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: 
            "10.1021/acschembio.7b00348"; journalName: "ACS Chem Biol"; date: 
            "2017-10-20- 20"; volume: "12"; issue: "10"; pages: "2510-2514"
COMMENT     Sequence Label: pRSETA_ionRD
FEATURES             Location/Qualifiers
     source          1..4458
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             103..120
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1395..1406
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(1859..1877)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1873..1920
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2017..2472
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2104..2123)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2314..2335
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2498..2602
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2603..2671,2672..3463)
                     /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     complement(2821..2840)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3634..4222
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4123..4142
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4376..4393
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcatcatca tcaccatcat
      121 gtgagcaaag gtgaagaact gtttaccggt gttgttccga ttctggttga actggatggt
      181 gacgttaatg gtcacaaatt ttcagttagc ggtgaaggcg aaggtgatgc aacctatggt
      241 aaactgaccc tgaaatttat ctgtaccacc ggcaaactgc cggttccgtg gccgaccctg
      301 gttaccaccc tgagctgggg tgttcagtgt tttgcacgtt atccggatca catgaaacag
      361 cacgattttt tcaaaagcgc aatgccggaa ggttatgttc aagaacgtac catcttcttc
      421 aaagatgacg gcaactataa aacccgtgcc gaagttaaat ttgaaggtga taccctggtg
      481 aatcgcattg aactgaaagg catcgatttt aaagaggatg gtaatatcct gggccacaaa
      541 ctggaatata atgccattca tggcaacgtg tatatcaccg cagataaaca gaaaaacggc
      601 atcaaagcaa attttggcct gaactgcaat attgaagatg gttcagttca gctggcagat
      661 cattatcagc agaatacccc gattggtgat ggtccggttc tgctgccgga taatcattat
      721 ctgagcaccc agagcaaact gagcaaagat ccgaatgaaa aacgtgatca catggtgctg
      781 ctggaatttg ttaccgcagc aggtattacc ctgggtatgg atgaactgta taaagcagca
      841 gcagccgcac gtgcagccgc agcccgtgcc gctgcagctc gtgcggcagc agcacgcgca
      901 gctgctgctc gcgctgcagc agctcgtgca ggtagcggtg gtagtggtgg ttcaggtggc
      961 agcggtggct ctggtggctc aggcgcagat gcagcagcgg ctgatgccgc tgccgctgac
     1021 gcagctgccg cagatgccgc agctgcagat gcagccgcag cggatgctgc agccgcagct
     1081 tccatggtgt caaaaggcga ggaactgttc acaggcgtgg tgccgatcct ggtagagctg
     1141 gacggggatg tgaatggcca taaattcagc gtttcaggtg aaggtgaggg cgacgccacg
     1201 tacggaaaac tgacactgaa attcatttgc acaacaggta aactgcctgt gccttggcct
     1261 acactggtga ccacctttgg ttatggtctg atgtgctttg ctcgctatcc tgaccacatg
     1321 aaacaacatg atttctttaa atctgccatg cctgaaggct acgtgcaaga gcgcaccatt
     1381 tttttcaaag acgatgggaa ttacaaaaca cgtgcggagg tgaaattcga gggcgataca
     1441 ctggttaacc gtatcgagct gaaaggtatc gacttcaaag aggacggaaa cattctgggt
     1501 cataaactgg aatacaacta caacagccat aacgtgtaca tcatggccga caaacaaaaa
     1561 aacgggatta aagtgaactt caaaatccgc cacaacatcg aagatggcag cgtgcagctg
     1621 gccgaccact atcaacaaaa cacaccgatc ggcgacggtc ctgtactgct gcctgacaac
     1681 cactatctgt catatcagag cgcactgtca aaagatccta acgagaaacg cgaccacatg
     1741 gttctgctgg aattcgtgac agccgctggc attacactgg gcatggacga gctgtacaaa
     1801 taaaagcttg atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     1861 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     1921 ctgaaaggag gaactatatc cggatctggc gtaatagcga agaggcccgc accgatcgcc
     1981 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     2041 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     2101 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     2161 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     2221 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     2281 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     2341 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     2401 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     2461 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2521 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2581 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2641 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2701 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2761 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     2821 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     2881 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     2941 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3001 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3061 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3121 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3181 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3241 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3301 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3361 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3421 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3481 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3541 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3601 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3661 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3721 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3781 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3841 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3901 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3961 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4021 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4081 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4141 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4201 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4261 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4321 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4381 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     4441 gccgattcat taatgcag
//
