LOCUS       sequence-111221-        6085 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET biosensor Destination vector for E. coli expression. Encodes a 
            FRET pair for fusion with ligand binding domain of interest..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6085)
  AUTHORS   Jones AM, Danielson JA, Manojkumar SN, Lanquar V, Grossmann G, 
            Frommer WB
  TITLE     Abscisic acid dynamics in roots detected with genetically encoded 
            FRET sensors.
  JOURNAL   eLife. 2014 Apr 15;3:e01741. doi: 10.7554/eLife.01741.
  PUBMED    24737862
REFERENCE   2  (bases 1 to 6085)
  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: "eLife."; 
            date: "2014-04-15"; volume: "3:e01741. doi"; pages: " 
            10.7554/eLife.01741"
COMMENT     Sequence Label: pFLIP34
FEATURES             Location/Qualifiers
     source          1..6085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..38
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        19..37
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             69..91
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             111..128
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             132..164
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(168..176,177..191)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     protein_bind    906..1030
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1055..1060,1061..1078,1079..1085)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     primer_bind     complement(1205..1224)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2198..2503
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2382..2401
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2544..2668)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3342..3371
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3430..3448)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3444..3491
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3588..4043
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3675..3694)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3885..3906
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4124..4228
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4229..4297,4298..5089)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLID*MEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5260..5848
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5749..5768
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6002..6019
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta
       61 gaaataattt tgtttaactt taagaaggag atatacatat gcggggttct catcatcatc
      121 atcatcatgg tatggctagc atgactggtg gacagcaaat gggtcgggat ctgtacgacg
      181 atgacgataa ggatccgggc ctcgaggttt ctaaaggtga agaattgttt acgggcgtcg
      241 tcccgatcct cgtggaactc gacggggatg ttaacgggca taagttttcg gtcagcgggg
      301 aaggggaggg ggacgcgacg tatgggaagc tcactctcaa gctgatctgt acgacgggga
      361 aactcccggt cccgtggccg acgctggtca cgacgctggg atacgggctc caatgctttg
      421 cgaggtatcc ggaccacatg aaacagcatg actttttcaa atcggcgatg ccggagggat
      481 acgtgcagga acggacgatc tttttcaaag acgatgggaa ctataagacg cgggcggaag
      541 tcaagtttga aggggacacg ctcgtcaacc ggatcgaact caaggggatt gacttcaaag
      601 aggatgggaa catactcggc cataagctcg aatacaatta caactcgcat aacgtataca
      661 tcaccgcgga taagcaaaag aacgggatca aagccaattt caaaatccgg cataacatag
      721 aggatggggg ggtccaactg gcggatcact atcagcaaaa cacgccgata ggggatgggc
      781 cggtcctcct cccggataac cattacctct cgtaccaaag cgcgctctcg aaggacccga
      841 atgagaaacg ggaccacatg gttctcctgg agttcgtcac ggcggcgggc ataggtaccg
      901 atatcacaag tttgtacaaa aaagctgaac gagaaacgta aaatgatata aatatcaata
      961 tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatcc
     1021 agtcactatg gcggccgcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     1081 taatgtgtgg attttgagtt aggatccgtc gagattttca ggagctaagg aagctaaaat
     1141 ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca
     1201 ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat
     1261 tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca
     1321 cattcttgcc cgcctgatga atgctcatcc ggaattccgt atggcaatga aagacggtga
     1381 gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac
     1441 gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc
     1501 gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa
     1561 tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc
     1621 caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga
     1681 caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg ggcttccatg
     1741 tcggcagaat gcttaatgaa ttacacagta ctgcgatgag tggcagggcg gggcgtaaac
     1801 gcgtggatcc ggcttactaa aagccagata acagtatgcg tatttgcgcg ctgatttttg
     1861 cggtataaga atatatactg atatgtatac ccgaagtatg tcaaaaagag gtatgctatg
     1921 aagcagcgta ttacagtgac agttgacagc gacagctatc agttgctcaa ggcatatatg
     1981 atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa tgaagcccgt cgtctgcgtg
     2041 ccgaacgctg gaaagcggaa aatcaggaag ggatggctga ggtcgcccgg tttattgaaa
     2101 tgaacggctc ttttgctgac gagaacaggg gctggtgaaa tgcagtttaa ggtttacacc
     2161 tataaaactt ttgctgacga gaacaggggc tggtgaaatg cagtttaagg tttacaccta
     2221 taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc
     2281 cgggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtctcccg
     2341 tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat
     2401 ggccagtgtg ccggtctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa
     2461 tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag gctcccttat
     2521 acacagccag tctgcaggtc gaccatagtg actggatatg ttgtgtttta cagtattatg
     2581 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
     2641 ctcgttcagc tttcttgtac aaagtggtga tatcgactag taccacaatg ggcgtaatca
     2701 agcccgacat gaagatcaag ctgaagatgg agggcaacgt gaatggccac gccttcgtga
     2761 tcgagggcga gggcgagggc aagccctacg acggcaccaa caccatcaac ctggaggtga
     2821 aggagggagc ccccctgccc ttctcctacg acattctgac caccgcgttc gcctacggca
     2881 acagggcctt caccaagtac cccgacgaca tccccaacta cttcaagcag tccttccccg
     2941 agggctactc ttgggagcgc accatgacct tcgaggacaa gggcatcgtg aaggtgaagt
     3001 ccgacatctc catggaggag gactccttca tctacgagat acacctcaag ggcgagaact
     3061 tcccccccaa cggccccgtg atgcagaaga agaccaccgg ctgggacgcc tccaccgaga
     3121 ggatgtacgt gcgcgacggc gtgctgaagg gcgacgtcaa gcacaagctg ctgctggagg
     3181 gcggcggcca ccaccgcgtt gacttcaaga ccatctacag ggccaagaag gcggtgaagc
     3241 tgcccgacta tcactttgtg gaccaccgca tcgagatcct gaaccacgac aaggactaca
     3301 acaaggtgac cgtttacgag agcgccgtgg cccgcaactc cgaacaaaaa ttgataagtg
     3361 aggaagattt ataagggccc gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     3421 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
     3481 ggggtttttt gctgaaagga ggaactatat ccggatctgg cgtaatagcg aagaggcccg
     3541 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     3601 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3661 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3721 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagag ctttacggca
     3781 cctcgaccgc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     3841 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3901 aactggaaca acactcaacc ctatcgcggt ctattctttt gatttataag ggattttgcc
     3961 gatttcggcc tattggttaa aaaatgagct gatttaacaa atatttaacg cgaattttaa
     4021 caaaatatta acgtttacaa tttcgcctga tgcggtattt tctccttacg catctgtgcg
     4081 gtatttcaca ccgcatacag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4141 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4201 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4261 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4321 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4381 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4441 agttctgcta tgtgatacac tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4501 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4561 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4621 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4681 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4741 accaaacgac gagagtgaca ccacgatgcc tgtagcaatg ccaacaacgt tgcgcaaact
     4801 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact gaatggaggc
     4861 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4921 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4981 taagcgctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5041 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5101 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5161 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5221 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5281 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5341 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5401 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5461 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5521 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5581 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5641 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5701 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5761 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5821 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5881 gggcttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5941 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6001 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     6061 gcgttggccg attcattaat gcagg
//
