LOCUS       sequence-221621-        6490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Endoplasmic Reticulum, Excitation: 559, Emission: 600.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6490)
  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 6490)
  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-Calnexin-N-14
FEATURES             Location/Qualifiers
     source          1..6490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2377..2386
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2383..3093
                     /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    3218..3339
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3255..3274)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3309..3328
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3346..3801)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3483..3504)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3695..3714
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3828..3932
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3934..4291
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3934..4125
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3957..3977)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4142..4277
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4326..5120
                     /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             4326..5120
                     /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(4380..4399)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4990..5009
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5308..5327)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5352..5399
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5728..6316
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6217..6236
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagac cgccatggaa gggaagtggt tgctgtgtat gttactggtg
      601 cttggaactg ctattgttga ggctcatgat ggacatgatg atgatgtgat tgatattgag
      661 gatgaccttg acgatgtcat tgaagaggta gaagactcaa aaccagatac cactgctcct
      721 ccttcatctc ccaaggttac ttacaaagct ccagttccaa caggggaagt atattttgct
      781 gattcttttg acagaggaac tctgtcaggg tggattttat ccaaagccaa gaaagacgat
      841 accgatgatg aaattgccaa atatgatgga aagtgggagg tagaggaaat gaaggagtca
      901 aagcttccag gtgataaagg acttgtgttg atgtctcggg ccaagcatca tgccatctct
      961 gctaaactga acaagccctt cctgtttgac accaagcctc tcattgttca gtatgaggtt
     1021 aatttccaaa atggaataga atgtggtggt gcctatgtga aactgctttc taaaacacca
     1081 gaactcaacc tggatcagtt ccatgacaag accccttata cgattatgtt tggtccagat
     1141 aaatgtggag aggactataa actgcacttc atcttccgac acaaaaaccc caaaacgggt
     1201 atctatgaag aaaaacatgc taagaggcca gatgcagatc tgaagaccta ttttactgat
     1261 aagaaaacac atctttacac actaatcttg aatccagata atagttttga aatactggtt
     1321 gaccaatctg tggtgaatag tggaaatctg ctcaatgaca tgactcctcc tgtaaatcct
     1381 tcacgtgaaa ttgaggaccc agaagaccgg aagcccgagg attgggatga aagaccaaaa
     1441 atcccagatc cagaagctgt caagccagat gactgggatg aagatgcccc tgctaagatt
     1501 ccagatgaag aggccacaaa acccgaaggc tggttagatg atgagcctga gtacgtacct
     1561 gatccagacg cagagaaacc tgaggattgg gatgaagaca tggatggaga atgggaggct
     1621 cctcagattg ccaaccctag atgtgagtca gctcctggat gtggtgtctg gcagcgacct
     1681 gtgattgaca accccaatta taaaggcaaa tggaagcctc ctatgattga caatcccagt
     1741 taccagggaa tctggaaacc caggaaaata ccaaatccag atttctttga agatctggaa
     1801 cctttcagaa tgactccttt tagtgctatt ggtttggagc tgtggtccat gacctctgac
     1861 attttttttg acaactttat catttgtgct gatcgaagaa tagttgatga ttgggccaat
     1921 gatggatggg gcctgaagaa agctgctgat ggggctgctg agccaggcgt tgtggggcag
     1981 atgaacgagg cagctgaaga gcgcccgtgg ctgtgggtag tctatattct aactgtagcc
     2041 cttcctgtgt tcctggttat cctcttctgc tgttctggaa agaaacagac cagtggtatg
     2101 gagtataaga aaactgatgc acctcaaccg gatgtgaagg aagaggaaga agagaaggaa
     2161 gaggaaaagg acaagggaga tgaggaggag gaaggagaag agaaacttga agagaaacag
     2221 aaaagtgatg ctgaagaaga tggtggcact gtcagtcaag aggaggaaga cagaaaacct
     2281 aaagcagagg aggatgaaat tttgaacaga tcaccaagaa acagaaagcc acgaagagag
     2341 gggggtgcgt caggcggcag cggatctgga ccggtcgcca ccatggtgtc taagggcgaa
     2401 gagctgatca aggaaaatat gcgtatgaag gtggtcatgg aaggttcggt caacggccac
     2461 caattcaaat gcacaggtga aggagaaggc aatccgtaca tgggaactca aaccatgagg
     2521 atcaaagtca tcgagggagg acccctgcca tttgcctttg acattcttgc cacgtcgttc
     2581 atgtatggca gccgtacttt tatcaagtac ccgaaaggca ttcctgattt ctttaaacag
     2641 tcctttcctg agggttttac ttgggaaaga gttacgagat acgaagatgg tggagtcgtc
     2701 accgtcatgc aggacaccag ccttgaggat ggctgtctcg tttaccacgt ccaagtcaga
     2761 ggggtaaact ttccctccaa tggtcccgtg atgcagaaga agaccaaggg ttgggagcct
     2821 aatacagaga tgatgtatcc agcagatggt ggtctgaggg gatacactca tatggcactg
     2881 aaagttgatg gtggtggcca tctgtcttgc tctttcgtaa caacttacag gtcaaaaaag
     2941 accgtcggga acatcaagat gcccggtatc catgccgttg atcaccgcct ggaaaggtta
     3001 gaggaaagtg acaatgaaat gttcgtagta caacgcgaac acgcagttgc caagttcgcc
     3061 gggcttggtg gtgggatgga cgagctgtac aagtaagcgg ccgcgactct agatcataat
     3121 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     3181 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     3241 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3301 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     3361 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3421 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3481 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3541 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3601 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3661 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3721 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3781 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3841 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3901 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     3961 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4021 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4081 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4141 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4201 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4261 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     4321 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4381 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4441 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4501 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4561 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4621 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4681 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4741 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4801 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     4861 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4921 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4981 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5041 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5101 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     5161 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5221 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5281 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     5341 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     5401 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     5461 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     5521 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     5581 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5641 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5701 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5761 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5821 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5881 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5941 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6001 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6061 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6121 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6181 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6241 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6301 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6361 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6421 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     6481 tggagttccg
//
