LOCUS       sequence-393234-        6717 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Nuclear Envelope, Excitation: 505, Emission: 515.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6717)
  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 6717)
  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: Clover-LaminA-C-18
FEATURES             Location/Qualifiers
     source          1..6717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             148..942
                     /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             148..942
                     /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(202..221)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     812..831
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1130..1149)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1174..1221
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1550..2138
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2039..2058
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2313..2616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2617..2820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      2859..2868
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2865..3581
                     /codon_start=1
                     /product="monomeric derivative of the bright green-yellow 
                     fluorescent protein Clover"
                     /label=mClover2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2910..2931)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     3518..3539
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5757..5878
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5794..5813)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5848..5867
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5885..6340)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6022..6043)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6234..6253
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6367..6471
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6473..6717,1..113)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6473..6664
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6496..6516)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6681..6717,1..99)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
       61 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
      121 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
      181 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
      241 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
      301 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
      361 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
      421 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
      481 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
      541 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
      601 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
      661 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
      721 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
      781 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
      841 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
      901 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
      961 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     1021 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     1081 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     1141 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     1201 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     1261 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     1321 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     1381 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     1441 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     1501 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1561 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1621 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1681 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     1741 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     1801 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     1861 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     1921 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     1981 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2041 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2101 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2161 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     2221 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg
     2281 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     2341 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     2401 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     2461 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     2521 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     2581 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     2641 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     2701 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     2761 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     2821 ggtttagtga accgtcagat ccgctagcgc taccggtcgc caccatggtg agcaagggcg
     2881 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2941 acaagttcag cgtccgcggc gagggcgagg gcgatgccac caacggcaag ctgaccctga
     3001 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg
     3061 gctacggcgt ggcctgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     3121 agtccgccat gcccgaaggc tacgtccagg agcgcaccat ctctttcaag gacgacggta
     3181 cctacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     3241 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     3301 tcaacagcca caacgtctat atcacggccg acaagcagaa gaacggcatc aaggctaact
     3361 tcaagatccg ccacaacgtt gaggacggca gcgtgcagct cgccgaccac taccagcaga
     3421 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agccatcagt
     3481 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     3541 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtccggactc agatctggca
     3601 gcggtggagg cagcgcatcc ggcggaagcg gaagcgagac cccgtcccag cggcgcgcca
     3661 cccgcagcgg ggcgcaggcc agctccactc cgctgtcgcc cacccgcatc acccggctgc
     3721 aggagaagga ggacctgcag gagctcaatg atcgcttggc ggtctacatc gaccgtgtgc
     3781 gctcgctgga aacggagaac gcagggctgc gccttcgcat caccgagtct gaagaggtgg
     3841 tcagccgcga ggtgtccggc atcaaggccg cctacgaggc cgagctcggg gatgcccgca
     3901 agacccttga ctcagtagcc aaggagcgcg cccgcctgca gctggagctg agcaaagtgc
     3961 gtgaggagtt taaggagctg aaagcgcgca ataccaagaa ggagggtgac ctgatagctg
     4021 ctcaggctcg gctgaaggac ctggaggctc tgctgaactc caaggaggcc gcactgagca
     4081 ctgctctcag tgagaagcgc acgctggagg gcgagctgca tgatctgcgg ggccaggtgg
     4141 ccaagcttga ggcagcccta ggtgaggcca agaagcaact tcaggatgag atgctgcggc
     4201 gggtggatgc tgagaacagg ctgcagacca tgaaggagga actggacttc cagaagaaca
     4261 tctacagtga ggagctgcgt gagaccaagc gccgtcatga gacccgactg gtggagattg
     4321 acaatgggaa gcagcgtgag tttgagagcc ggctggcgga tgcgctgcag gaactgcggg
     4381 cccagcatga ggaccaggtg gagcagtata agaaggagct ggagaagact tattctgcca
     4441 agctggacaa tgccaggcag tctgctgaga ggaacagcaa cctggtgggg gctgcccacg
     4501 aggagctgca gcagtcgcgc atccgcatcg acagcctctc tgcccagctc agccagctcc
     4561 agaagcagct ggcagccaag gaggcgaagc ttcgagacct ggaggactca ctggcccgtg
     4621 agcgggacac cagccggcgg ctgctggcgg aaaaggagcg ggagatggcc gagatgcggg
     4681 caaggatgca gcagcagctg gacgagtacc aggagcttct ggacatcaag ctggccctgg
     4741 acatggagat ccacgcctac cgcaagctct tggagggcga ggaggagagg ctacgcctgt
     4801 cccccagccc tacctcgcag cgcagccgtg gccgtgcttc ctctcactca tcccagacac
     4861 agggtggggg cagcgtcacc aaaaagcgca aactggagtc cactgagagc cgcagcagct
     4921 tctcacagca cgcacgcact agcgggcgcg tggccgtgga ggaggtggat gaggagggca
     4981 agtttgtccg gctgcgcaac aagtccaatg aggaccagtc catgggcaat tggcagatca
     5041 agcgccagaa tggagatgat cccttgctga cttaccggtt cccaccaaag ttcaccctga
     5101 aggctgggca ggtggtgacg atctgggctg caggagctgg ggccacccac agccccccta
     5161 ccgacctggt gtggaaggca cagaacacct ggggctgcgg gaacagcctg cgtacggctc
     5221 tcatcaactc cactggggaa gaagtggcca tgcgcaagct ggtgcgctca gtgactgtgg
     5281 ttgaggacga cgaggatgag gatggagatg acctgctcca tcaccaccac ggctcccact
     5341 gcagcagctc gggggacccc gctgagtaca acctgcgctc gcgcaccgtg ctgtgcggga
     5401 cctgcgggca gcctgccgac aaggcatctg ccagcggctc aggagcccag gtgggcggac
     5461 ccatctcctc tggctcttct gcctccagtg tcacggtcac tcgcagctac cgcagtgtgg
     5521 ggggcagtgg gggtggcagc ttcggggaca atctggtcac ccgctcctac ctcctgggca
     5581 actccagccc ccgaacccag agcccccaga actgcagcat catgtaagga tccaccggat
     5641 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     5701 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     5761 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5821 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac
     5881 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     5941 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     6001 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     6061 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     6121 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     6181 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     6241 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     6301 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     6361 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     6421 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     6481 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     6541 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     6601 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     6661 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctc
//
