LOCUS       sequence-296256-        6800 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses far-red FP mNeptune2 fused to Dronpa145N. Dronpa145N 
            oligomerizes under violet light (390-400nm) and dissociates under 
            cyan light (490-500nm).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6800)
  AUTHORS   Zhou XX, Chung HK, Lam AJ, Lin MZ
  TITLE     Optical control of protein activity by fluorescent protein domains.
  JOURNAL   Science. 2012 Nov 9;338(6108):810-4. doi: 10.1126/science.1226854.
  PUBMED    23139335
REFERENCE   2  (bases 1 to 6800)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2012-11-9- 9"; volume: "338"; issue: 
            "6108"; pages: "810-4"
COMMENT     Sequence Label: pcDNA3-mNeptune2-N
FEATURES             Location/Qualifiers
     source          1..6800
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(40..60)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        45..374
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      225..360
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     287..306
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             441..1235
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(495..514)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1105..1124
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1409..1530
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1446..1465)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1500..1519
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1579..1595)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1579..1595)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1592..1614)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1603..1619
                     /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(1627..1633,1634..1651,1652..1657))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1672..1693
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1810..1827)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1981..2569)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2061..2080)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2740..3531,3532..3600))
                     /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     3363..3382
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3601..3705)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3780..3799)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3971..4350
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4351..4554
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4505..4525
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4599..4618
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4599..4617
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4631..5362
                     /codon_start=1
                     /product="far-red fluorescent protein with spectra similar 
                     to those of mNeptune but improved brightness"
                     /label=mNeptune2
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTG
                     RIKVVEGGPLPFAFDILATCFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTVTQDTSLQDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRCD
                     MALKLVGGGHLHCNLKTTYRSKKPAKNLKMPGVYFVDRRLERIKEADNETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     promoter        complement(6089..6107)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6090..6107)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6127..6144)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6133..6357
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6403..6800,1..31)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6490..6509)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6700..6721
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
       61 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
      121 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
      181 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
      241 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
      301 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
      361 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
      421 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
      481 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
      541 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
      601 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
      661 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
      721 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
      781 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
      841 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
      901 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
      961 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     1021 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     1081 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     1141 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     1201 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     1261 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     1321 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     1381 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     1441 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     1501 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     1561 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     1621 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     1681 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1741 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1801 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     1861 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     1921 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1981 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2041 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2101 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2161 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2221 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2281 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2341 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2401 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2461 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2521 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2581 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2641 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2701 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2761 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2821 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2881 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2941 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3001 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3061 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3121 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3181 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3241 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3301 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3361 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3421 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3481 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3541 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3601 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3661 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3721 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     3781 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     3841 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     3901 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     3961 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     4021 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     4081 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     4141 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     4201 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     4261 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     4321 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     4381 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     4441 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     4501 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
     4561 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
     4621 acccaagctt atggtgtcta agggcgaaga gctgattaag gagaacatgc acatgaagct
     4681 gtacatggag ggcaccgtga acaaccacca cttcaagtgc acatccgagg gcgaaggcaa
     4741 gccctacgag ggcacccaga ccggcagaat caaggtggtc gagggcggcc ctctcccctt
     4801 cgccttcgac atcctggcta cctgcttcat gtacggcagc aagaccttca tcaaccacac
     4861 ccagggcatc cccgatttct ttaagcagtc cttccctgag ggcttcacat gggagagagt
     4921 caccacatac gaagacgggg gcgtgcttac cgttacccag gacaccagcc tccaggacgg
     4981 ctgcttgatc tacaacgtca agctcagagg ggtgaacttc ccatccaacg gccctgtgat
     5041 gcagaagaaa acactcggct gggaggccag taccgagacc ctgtaccccg ctgacggcgg
     5101 cctggaaggc agatgcgaca tggccctgaa gctcgtgggc gggggccacc tgcactgcaa
     5161 cctgaagacc acatacagat ccaagaaacc cgctaagaac ctcaagatgc ccggcgtcta
     5221 ctttgtggac cgcagactgg aaagaatcaa ggaggccgac aatgagacct acgtcgagca
     5281 gcacgaggtg gctgtggcca gatactgcga cctccctagc aaactggggc acaaacttaa
     5341 tggcatggac gagctgtaca agggacttaa gggatccgaa ttcggcatga gcgtgatcaa
     5401 gcccgacatg aaaatcaagc tgaggatgga gggtgccgtg aacggtcacc cgtttgcgat
     5461 cgagggcgtg ggactgggca agccgttcga gggaaagcaa agcatggacc tcaaggtgaa
     5521 ggagggcgga ccgctgccct tcgcctacga cattctgacc actgtcttct gctacggtaa
     5581 tcgcgtcttc gcaaagtatc ccgaaaacat cgtggattac tttaagcaga gcttcccgga
     5641 gggttactcc tgggagcgat ccatgaacta cgaggacgga gggatctgca acgcgacgaa
     5701 cgatataacc ttggacggcg actgctacat ttacgaaatt cggttcgacg gcgtgaattt
     5761 cccagcgaat ggccctgtaa tgcaaaaacg cacggtaaag tgggagccca gcaccgagaa
     5821 cctgtacgtc cgcgacggag tcctgaaggg agacgtgaac atggcgcttt cattggaagg
     5881 tggcgggcac tacagatgcg atttcaagac gacctataag gcgaaaaagg tggtccaact
     5941 gccggactac catttcgtcg accaccacat cgagatcaag agccacgaca aagactatag
     6001 caacgtgaat ctccacgagc atgccgaggc ccacagcgag ctgcccaggc aggccaagta
     6061 actcgagcat gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct
     6121 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     6181 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     6241 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     6301 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     6361 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     6421 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     6481 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     6541 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     6601 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     6661 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     6721 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     6781 cggcctattg gttaaaaaat
//
