LOCUS       sequence-238453-        5386 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Cytosolic FRET, Excitation: , Emission: .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5386)
  AUTHORS   Shaner NC, Lambert GG, Chammas A, Ni Y, Cranfill PJ, Baird MA, Sell 
            BR, Allen JR, Day RN, Israelsson M, Davidson MW, Wang J
  TITLE     A bright monomeric green fluorescent protein derived from 
            Branchiostoma lanceolatum.
  JOURNAL   Nat Methods. 2013 May;10(5):407-9. doi: 10.1038/nmeth.2413. Epub 
            2013 Mar 24.
  PUBMED    23524392
REFERENCE   2  (bases 1 to 5386)
  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: 
            "10.1038/nmeth.2413"; journalName: "Nat Methods"; date: "2013-05"; 
            volume: "10"; issue: "5"; pages: "407-9"
COMMENT     Sequence Label: mNeonGreen-mRuby2-FRET-10
FEATURES             Location/Qualifiers
     source          1..5386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        109..412
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        413..616
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     567..587
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      643..652
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             649..1359
                     /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"
     regulatory      1384..1393
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1390..2100
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      1645..1654
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2222..2343
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2259..2278)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2313..2332
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2350..2805)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     2699..2718
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2832..2936
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2938..3295
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2938..3129
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2961..2981)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3146..3281
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3330..4124
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3330..4124
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3384..3403)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3994..4013
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4312..4331)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4356..4403
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4732..5320
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5221..5240
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata
       61 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      121 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      181 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      241 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      301 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      361 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      421 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      481 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      541 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      601 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagccaccat ggtgtctaag
      661 ggcgaagagc tgatcaagga aaatatgcgt atgaaggtgg tcatggaagg ttcggtcaac
      721 ggccaccaat tcaaatgcac aggtgaagga gaaggcaatc cgtacatggg aactcaaacc
      781 atgaggatca aagtcatcga gggaggaccc ctgccatttg cctttgacat tcttgccacg
      841 tcgttcatgt atggcagccg tacttttatc aagtacccga aaggcattcc tgatttcttt
      901 aaacagtcct ttcctgaggg ttttacttgg gaaagagtta cgagatacga agatggtgga
      961 gtcgtcaccg tcatgcagga caccagcctt gaggatggct gtctcgttta ccacgtccaa
     1021 gtcagagggg taaactttcc ctccaatggt cccgtgatgc agaagaagac caagggttgg
     1081 gagcctaata cagagatgat gtatccagca gatggtggtc tgaggggata cactcatatg
     1141 gcactgaaag ttgatggtgg tggccatctg tcttgctctt tcgtaacaac ttacaggtca
     1201 aaaaagaccg tcgggaacat caagatgccc ggtatccatg ccgttgatca ccgcctggaa
     1261 aggttagagg aaagtgacaa tgaaatgttc gtagtacaac gcgaacacgc agttgccaag
     1321 ttcgccgggc ttggtggtgg gatggacgag ctgtacaagt ccggactcag atccccaccg
     1381 gtcgccacca tggtgagcaa gggcgaggag gataacatgg cctctctccc agcgacacat
     1441 gagttacaca tctttggctc catcaacggt gtggactttg acatggtggg tcagggcacc
     1501 ggcaatccaa atgatggtta tgaggagtta aacctgaagt ccaccaaggg tgacctccag
     1561 ttctccccct ggattctggt ccctcatatc gggtatggct tccatcagta cctgccctac
     1621 cctgacggga tgtcgccttt ccaggccgcc atggtagatg gctccggcta ccaagtccat
     1681 cgcacaatgc agtttgaaga tggtgcctcc cttactgtta actaccgcta cacctacgag
     1741 ggaagccaca tcaaaggaga ggcccaggtg aaggggactg gtttccctgc tgacggtcct
     1801 gtgatgacca actcgctgac cgctgcggac tggtgcaggt cgaagaagac ttaccccaac
     1861 gacaaaacca tcatcagtac ctttaagtgg agttacacca ctggaaatgg caagcgctac
     1921 cggagcactg cgcggaccac ctacaccttt gccaagccaa tggcggctaa ctatctgaag
     1981 aaccagccga tgtacgtgtt ccgtaagacg gagctcaagc actccaagac cgagctcaac
     2041 ttcaaggagt ggcaaaaggc ctttaccgat gtgatgggca tggacgagct gtacaagtaa
     2101 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2161 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2221 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2281 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2341 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     2401 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     2461 agaccgagat agggttgagt gttgttccag ttcggaacaa gagtccacta ttaaagaacg
     2521 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     2581 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     2641 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     2701 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     2761 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     2821 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2881 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     2941 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3001 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3061 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3121 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3181 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3241 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     3301 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3361 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3421 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3481 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     3541 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3601 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3661 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3721 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3781 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     3841 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     3901 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     3961 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4021 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4081 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4141 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4201 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4261 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     4321 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     4381 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4441 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4501 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4561 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     4621 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4681 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4741 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4801 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4861 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4921 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4981 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5041 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5101 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5161 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5221 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5281 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5341 tttacggttc ctggcctttt gctggccttt tgctcacatg ttcttt
//
