LOCUS       sequence-136841-        7622 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nabi2.226 is a FRET (Forster Resonance Energy Transfer)-based 
            protein voltage sensor. This probe contains Clover and mRuby2 
            inserted at different locations in the Ciona voltage sensitive 
            domain..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7622)
  AUTHORS   Sung U, Sepehri-Rad M, Piao HH, Jin L, Hughes T, Cohen LB, Baker BJ
  TITLE     Developing Fast Fluorescent Protein Voltage Sensors by Optimizing 
            FRET Interactions.
  JOURNAL   PLoS One. 2015 Nov 20;10(11):e0141585. doi: 
            10.1371/journal.pone.0141585. eCollection 2015.
  PUBMED    26587834
REFERENCE   2  (bases 1 to 7622)
  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: "PLoS One.";
            date: "2015-11-20"; volume: "10(11):e0141585. doi"; pages: " 
            10.1371/journal.pone.0141585. eCollection 2015"
COMMENT     Sequence Label: Nabi2.226
FEATURES             Location/Qualifiers
     source          1..7622
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1220..1241)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1828..1849
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2324..3034
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     primer_bind     complement(3047..3064)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3053..3277
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3323..3751
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3410..3429)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3620..3641
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3760..3780)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3765..4094
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3945..4080
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4007..4026
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4143..5168
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4143..5168
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    5298..5419
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5335..5354)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5389..5408
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5468..5484)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5468..5484)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5481..5503)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5492..5508
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5516..5522,5523..5540,5541..5546))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5561..5582
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5699..5716)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5870..6455)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5950..5969)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6626..7417,7418..7486))
                     /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     7249..7268
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7487..7591)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttatggag ggattcgacg gttcagattt tagtcctcca gctgatttag ttggcgttga
      961 cggtgcagtc atgcggaacg tcgttgacgt cacgataaat ggtgacgtca ctgctccgcc
     1021 gaaagctgcg ccaagaaaaa gtgaatcggt aaagaaagtt cattggaatg atgtagacca
     1081 aggaccgagt gaaaaaccag agacaagaca ggaggaacga atagatatac ccgagatttc
     1141 aggtctatgg tggggcgaga atgaacatgg agtgatggtg agcaagggcg aggagctgtt
     1201 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1261 cgtccgcggc gagggcgagg gcgatgccac caacggcaag ctgaccctga agttcatctg
     1321 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg gctacggcgt
     1381 ggcctgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1441 gcccgaaggc tacgtccagg agcgcaccat ctctttcaag gacgacggta cctacaagac
     1501 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1561 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact tcaacagcca
     1621 caacgtctat atcacggccg acaagcagaa gaacggcatc aaggctaact tcaagatccg
     1681 ccacaacgtt gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     1741 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agccatcagt ccgccctgag
     1801 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccga
     1861 cgatgggaga atggagatac ctactactgg tgtaggtcgc gtccagtttc gtgtccgagc
     1921 agtgattgat catctaggga tgcgagtctt tggagtcttc ctaattttct tggacatcat
     1981 cctcatgatc attgatctca gtcttccagg aaaaagtgaa tcttcacaat ccttttatga
     2041 cgggatggct ttggctcttt cttgttattt catgctggat ttaggattaa ggatatttgc
     2101 ctacgggccc aagaatttct tcaccaaccc ctgggaggtt gctgatggtt tgattatcgt
     2161 tgtcacattc gtcgtcacga tattttacac tgtgttagat gaatacgtgc aagaaacagg
     2221 agccgatggt ttggggcaat tggttgtgtt ggcccgtttg ctgcgtgtgg ttcgattagc
     2281 aagaatattt tattcccacc aacaaatgaa ggcttcaagc agaatggtgt ctaagggcga
     2341 agagctgatc aaggaaaata tgcgtatgaa ggtggtcatg gaaggttcgg tcaacggcca
     2401 ccaattcaaa tgcacaggtg aaggagaagg caatccgtac atgggaactc aaaccatgag
     2461 gatcaaagtc atcgagggag gacccctgcc atttgccttt gacattcttg ccacgtcgtt
     2521 catgtatggc agccgtactt ttatcaagta cccgaaaggc attcctgatt tctttaaaca
     2581 gtcctttcct gagggtttta cttgggaaag agttacgaga tacgaagatg gtggagtcgt
     2641 caccgtcatg caggacacca gccttgagga tggctgtctc gtttaccacg tccaagtcag
     2701 aggggtaaac tttccctcca atggtcccgt gatgcagaag aagaccaagg gttgggagcc
     2761 taatacagag atgatgtatc cagcagatgg tggtctgagg ggatacactc atatggcact
     2821 gaaagttgat ggtggtggcc atctgtcttg ctctttcgta acaacttaca ggtcaaaaaa
     2881 gaccgtcggg aacatcaaga tgcccggtat ccatgccgtt gatcaccgcc tggaaaggtt
     2941 agaggaaagt gacaatgaaa tgttcgtagt acaacgcgaa cacgcagttg ccaagttcgc
     3001 cgggcttggt ggtgggatgg acgagctgta caagtaatag gcgcgccctc gactgtgcct
     3061 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3121 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3181 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3241 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3301 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3361 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3421 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3481 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3541 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3601 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3661 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3721 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3781 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3841 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3901 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3961 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4021 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4081 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg
     4141 tgatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg
     4201 acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg
     4261 atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag
     4321 atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca
     4381 ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt
     4441 tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg
     4501 atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag
     4561 gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt
     4621 atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg
     4681 agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg
     4741 gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg
     4801 cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg
     4861 cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc
     4921 cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg
     4981 acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg
     5041 gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg
     5101 gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa
     5161 aggaatagca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5221 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5281 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5341 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5401 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5461 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5521 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5581 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5641 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5701 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5761 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5821 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5881 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5941 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6001 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6061 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6121 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6181 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6241 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6301 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6361 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     6421 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6481 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6541 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6601 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6661 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6721 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6781 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6841 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6901 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6961 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7021 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7081 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7141 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7201 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7261 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7321 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7381 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7441 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7501 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7561 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7621 tc
//
