LOCUS       sequence-421916-        6711 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Nuclear Envelope, Excitation: 559, Emission: 600.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6711)
  AUTHORS   Lam AJ, St-Pierre F, Gong Y, Marshall JD, Cranfill PJ, Baird MA, 
            McKeown MR, Wiedenmann J, Davidson MW, Schnitzer MJ, Tsien RY, Lin 
            MZ
  TITLE     Improving FRET dynamic range with bright green and red fluorescent 
            proteins.
  JOURNAL   Nat Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171.
  PUBMED    22961245
REFERENCE   2  (bases 1 to 6711)
  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: "Nat 
            Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171."
COMMENT     Sequence Label: mRuby2-LaminA-C-18
FEATURES             Location/Qualifiers
     source          1..6711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             92..886
                     /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             92..886
                     /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(146..165)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     756..775
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1074..1093)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1118..1165
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1494..2082
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1983..2002
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2257..2560
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2561..2764
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      2803..2812
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2809..3519
                     /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"
     polyA_signal    5695..5816
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5732..5751)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5786..5805
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5823..6278)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5960..5981)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6172..6191
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6305..6409
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6411..6711,1..57)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6411..6602
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6434..6454)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6619..6711,1..43)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
       61 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
      121 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
      181 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
      241 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
      301 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
      361 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
      421 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
      481 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
      541 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
      601 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
      661 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
      721 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
      781 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
      841 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
      901 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
      961 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     1021 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     1081 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     1141 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     1201 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     1261 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     1321 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     1381 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     1441 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     1501 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     1561 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     1621 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     1681 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     1741 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     1801 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     1861 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     1921 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     1981 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2041 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2101 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2161 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     2221 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     2281 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     2341 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     2401 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     2461 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     2521 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     2581 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     2641 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     2701 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     2761 agctggttta gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat ggtgtctaag
     2821 ggcgaagagc tgatcaagga aaatatgcgt atgaaggtgg tcatggaagg ttcggtcaac
     2881 ggccaccaat tcaaatgcac aggtgaagga gaaggcaatc cgtacatggg aactcaaacc
     2941 atgaggatca aagtcatcga gggaggaccc ctgccatttg cctttgacat tcttgccacg
     3001 tcgttcatgt atggcagccg tacttttatc aagtacccga aaggcattcc tgatttcttt
     3061 aaacagtcct ttcctgaggg ttttacttgg gaaagagtta cgagatacga agatggtgga
     3121 gtcgtcaccg tcatgcagga caccagcctt gaggatggct gtctcgttta ccacgtccaa
     3181 gtcagagggg taaactttcc ctccaatggt cccgtgatgc agaagaagac caagggttgg
     3241 gagcctaata cagagatgat gtatccagca gatggtggtc tgaggggata cactcatatg
     3301 gcactgaaag ttgatggtgg tggccatctg tcttgctctt tcgtaacaac ttacaggtca
     3361 aaaaagaccg tcgggaacat caagatgccc ggtatccatg ccgttgatca ccgcctggaa
     3421 aggttagagg aaagtgacaa tgaaatgttc gtagtacaac gcgaacacgc agttgccaag
     3481 ttcgccgggc ttggtggtgg gatggacgag ctgtacaagt ccggactcag atctggcagc
     3541 ggtggaggca gcgcatccgg cggaagcgga agcgagaccc cgtcccagcg gcgcgccacc
     3601 cgcagcgggg cgcaggccag ctccactccg ctgtcgccca cccgcatcac ccggctgcag
     3661 gagaaggagg acctgcagga gctcaatgat cgcttggcgg tctacatcga ccgtgtgcgc
     3721 tcgctggaaa cggagaacgc agggctgcgc cttcgcatca ccgagtctga agaggtggtc
     3781 agccgcgagg tgtccggcat caaggccgcc tacgaggccg agctcgggga tgcccgcaag
     3841 acccttgact cagtagccaa ggagcgcgcc cgcctgcagc tggagctgag caaagtgcgt
     3901 gaggagttta aggagctgaa agcgcgcaat accaagaagg agggtgacct gatagctgct
     3961 caggctcggc tgaaggacct ggaggctctg ctgaactcca aggaggccgc actgagcact
     4021 gctctcagtg agaagcgcac gctggagggc gagctgcatg atctgcgggg ccaggtggcc
     4081 aagcttgagg cagccctagg tgaggccaag aagcaacttc aggatgagat gctgcggcgg
     4141 gtggatgctg agaacaggct gcagaccatg aaggaggaac tggacttcca gaagaacatc
     4201 tacagtgagg agctgcgtga gaccaagcgc cgtcatgaga cccgactggt ggagattgac
     4261 aatgggaagc agcgtgagtt tgagagccgg ctggcggatg cgctgcagga actgcgggcc
     4321 cagcatgagg accaggtgga gcagtataag aaggagctgg agaagactta ttctgccaag
     4381 ctggacaatg ccaggcagtc tgctgagagg aacagcaacc tggtgggggc tgcccacgag
     4441 gagctgcagc agtcgcgcat ccgcatcgac agcctctctg cccagctcag ccagctccag
     4501 aagcagctgg cagccaagga ggcgaagctt cgagacctgg aggactcact ggcccgtgag
     4561 cgggacacca gccggcggct gctggcggaa aaggagcggg agatggccga gatgcgggca
     4621 aggatgcagc agcagctgga cgagtaccag gagcttctgg acatcaagct ggccctggac
     4681 atggagatcc acgcctaccg caagctcttg gagggcgagg aggagaggct acgcctgtcc
     4741 cccagcccta cctcgcagcg cagccgtggc cgtgcttcct ctcactcatc ccagacacag
     4801 ggtgggggca gcgtcaccaa aaagcgcaaa ctggagtcca ctgagagccg cagcagcttc
     4861 tcacagcacg cacgcactag cgggcgcgtg gccgtggagg aggtggatga ggagggcaag
     4921 tttgtccggc tgcgcaacaa gtccaatgag gaccagtcca tgggcaattg gcagatcaag
     4981 cgccagaatg gagatgatcc cttgctgact taccggttcc caccaaagtt caccctgaag
     5041 gctgggcagg tggtgacgat ctgggctgca ggagctgggg ccacccacag cccccctacc
     5101 gacctggtgt ggaaggcaca gaacacctgg ggctgcggga acagcctgcg tacggctctc
     5161 atcaactcca ctggggaaga agtggccatg cgcaagctgg tgcgctcagt gactgtggtt
     5221 gaggacgacg aggatgagga tggagatgac ctgctccatc accaccacgg ctcccactgc
     5281 agcagctcgg gggaccccgc tgagtacaac ctgcgctcgc gcaccgtgct gtgcgggacc
     5341 tgcgggcagc ctgccgacaa ggcatctgcc agcggctcag gagcccaggt gggcggaccc
     5401 atctcctctg gctcttctgc ctccagtgtc acggtcactc gcagctaccg cagtgtgggg
     5461 ggcagtgggg gtggcagctt cggggacaat ctggtcaccc gctcctacct cctgggcaac
     5521 tccagccccc gaacccagag cccccagaac tgcagcatca tgtaaggatc caccggatct
     5581 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     5641 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     5701 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5761 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     5821 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     5881 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     5941 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     6001 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     6061 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     6121 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     6181 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     6241 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     6301 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     6361 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     6421 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     6481 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     6541 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     6601 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     6661 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc c
//
