LOCUS       sequence-136837-        7610 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nabi2.30 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 7610)
  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 7610)
  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: Nabi2.30
FEATURES             Location/Qualifiers
     source          1..7610
                     /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             2312..3022
                     /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(3035..3052)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3041..3265
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3311..3739
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3398..3417)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3608..3629
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3748..3768)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3753..4082
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3933..4068
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3995..4014
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4131..5156
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4131..5156
                     /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    5286..5407
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5323..5342)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5377..5396
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5456..5472)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5456..5472)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5469..5491)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5480..5496
                     /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(5504..5510,5511..5528,5529..5534))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5549..5570
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5687..5704)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5858..6443)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5938..5957)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6614..7405,7406..7474))
                     /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     7237..7256
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7475..7579)
                     /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 gatggtgtct aagggcgaag agctgatcaa
     2341 ggaaaatatg cgtatgaagg tggtcatgga aggttcggtc aacggccacc aattcaaatg
     2401 cacaggtgaa ggagaaggca atccgtacat gggaactcaa accatgagga tcaaagtcat
     2461 cgagggagga cccctgccat ttgcctttga cattcttgcc acgtcgttca tgtatggcag
     2521 ccgtactttt atcaagtacc cgaaaggcat tcctgatttc tttaaacagt cctttcctga
     2581 gggttttact tgggaaagag ttacgagata cgaagatggt ggagtcgtca ccgtcatgca
     2641 ggacaccagc cttgaggatg gctgtctcgt ttaccacgtc caagtcagag gggtaaactt
     2701 tccctccaat ggtcccgtga tgcagaagaa gaccaagggt tgggagccta atacagagat
     2761 gatgtatcca gcagatggtg gtctgagggg atacactcat atggcactga aagttgatgg
     2821 tggtggccat ctgtcttgct ctttcgtaac aacttacagg tcaaaaaaga ccgtcgggaa
     2881 catcaagatg cccggtatcc atgccgttga tcaccgcctg gaaaggttag aggaaagtga
     2941 caatgaaatg ttcgtagtac aacgcgaaca cgcagttgcc aagttcgccg ggcttggtgg
     3001 tgggatggac gagctgtaca agtaataggc gcgccctcga ctgtgccttc tagttgccag
     3061 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3121 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3181 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3241 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     3301 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3361 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3421 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3481 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3541 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3601 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3661 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3721 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3781 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3841 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3901 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3961 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4021 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4081 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg atgaaaaagc
     4141 ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg
     4201 acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc
     4261 gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt
     4321 atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca
     4381 gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc
     4441 ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg
     4501 cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat
     4561 acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa
     4621 ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt
     4681 gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg
     4741 tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg
     4801 attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc
     4861 agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg
     4921 cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg
     4981 atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg
     5041 tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag
     5101 tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagcacg
     5161 tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5221 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5281 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5341 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5401 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5461 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5521 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5581 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5641 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5701 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5761 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5821 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5881 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5941 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6001 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6061 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6121 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6181 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6241 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6301 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6361 cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta
     6421 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6481 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6541 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6601 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6661 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6721 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6781 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6841 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6901 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6961 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7021 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7081 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7141 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7201 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7261 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7321 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7381 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7441 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7501 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7561 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
