LOCUS       sequence-136838-        7643 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nabi2.104 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 7643)
  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 7643)
  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: Nabi2.104
FEATURES             Location/Qualifiers
     source          1..7643
                     /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             2345..3055
                     /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(3068..3085)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3074..3298
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3344..3772
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3431..3450)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3641..3662
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3781..3801)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3786..4115
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3966..4101
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4028..4047
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4164..5189
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4164..5189
                     /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    5319..5440
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5356..5375)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5410..5429
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5489..5505)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5489..5505)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5502..5524)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5513..5529
                     /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(5537..5543,5544..5561,5562..5567))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5582..5603
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5720..5737)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5891..6476)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5971..5990)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6647..7438,7439..7507))
                     /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     7270..7289
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7508..7612)
                     /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 agaagaacaa tatcacagaa
     2341 caagatggtg tctaagggcg aagagctgat caaggaaaat atgcgtatga aggtggtcat
     2401 ggaaggttcg gtcaacggcc accaattcaa atgcacaggt gaaggagaag gcaatccgta
     2461 catgggaact caaaccatga ggatcaaagt catcgaggga ggacccctgc catttgcctt
     2521 tgacattctt gccacgtcgt tcatgtatgg cagccgtact tttatcaagt acccgaaagg
     2581 cattcctgat ttctttaaac agtcctttcc tgagggtttt acttgggaaa gagttacgag
     2641 atacgaagat ggtggagtcg tcaccgtcat gcaggacacc agccttgagg atggctgtct
     2701 cgtttaccac gtccaagtca gaggggtaaa ctttccctcc aatggtcccg tgatgcagaa
     2761 gaagaccaag ggttgggagc ctaatacaga gatgatgtat ccagcagatg gtggtctgag
     2821 gggatacact catatggcac tgaaagttga tggtggtggc catctgtctt gctctttcgt
     2881 aacaacttac aggtcaaaaa agaccgtcgg gaacatcaag atgcccggta tccatgccgt
     2941 tgatcaccgc ctggaaaggt tagaggaaag tgacaatgaa atgttcgtag tacaacgcga
     3001 acacgcagtt gccaagttcg ccgggcttgg tggtgggatg gacgagctgt acaagtaata
     3061 ggcgcgccct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3121 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3181 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3241 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3301 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3361 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3421 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3481 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3541 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3601 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3661 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3721 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3781 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3841 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3901 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3961 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4021 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4081 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4141 ttttcggatc tgatcagcac gtgatgaaaa agcctgaact caccgcgacg tctgtcgaga
     4201 agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag
     4261 aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct
     4321 gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc
     4381 cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc
     4441 gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc
     4501 agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt
     4561 tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg
     4621 cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt
     4681 ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc
     4741 acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag
     4801 cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct
     4861 tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc
     4921 atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc
     4981 aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat
     5041 gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa
     5101 gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc
     5161 ccagcactcg tccgagggca aaggaatagc acgtgctacg agatttcgat tccaccgccg
     5221 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5281 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5341 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5401 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5461 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5521 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5581 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5641 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5701 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5761 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5821 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5881 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5941 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6001 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6061 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6121 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6181 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6241 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6301 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6361 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6421 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6481 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6541 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6601 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6661 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6721 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6781 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6841 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6901 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6961 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7021 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7081 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7141 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7201 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7261 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7321 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7381 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7441 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7501 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7561 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7621 cccgaaaagt gccacctgac gtc
//
