LOCUS       sequence-173191-        5221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Nucleus/Centromeres, Excitation: 559, Emission: 600.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5221)
  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 5221)
  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-CENPB-N-22
FEATURES             Location/Qualifiers
     source          1..5221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    508..560
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      568..577
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             574..1284
                     /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    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"
     rep_origin      complement(1537..1992)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1674..1695)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1886..1905
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2019..2123
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2125..2482
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2125..2316
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2148..2168)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2333..2468
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2517..3311
                     /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             2517..3311
                     /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(2571..2590)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3181..3200
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3499..3518)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3543..3590
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3919..4507
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4408..4427
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4682..4985
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4986..5189
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5140..5160
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5216..5221,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atgggcccca agcggcggca gctgacgttc cgggagaagt cgcggatcat ccaggaggtg
       61 gaggagaacc cggacctacg caagggcgag atcgcgcggc gcttcaacat cccgccgtcc
      121 acgctgagca cgatcctgaa gaacaagcgc gccatcctgg cgtcggagcg caagtacggg
      181 gtggcctcca cctgccgcaa gaccaacaag ctgtctccct acgacaagct cgagggcttg
      241 ctcatcgcct ggttccagca gatccgcgcc gccggcctgc cggtcaaggg catcatcctc
      301 aaggagaagg cgctgcgcat agccgaggag ctgggcatgg acgacttcac cgcctccaac
      361 ggctggctgg accgcttccg ccggcgccac ggcgtggtgt cctgcagcgg cgtggcccgc
      421 gcccgcgcgc gaaacgctgc cccccgcacc ccggcggcgc ctgccagtcc ggccgcggtg
      481 ccctcggagg gcagtggcgg gagcactctc gagctcaagc ttcgaattct gcagtcgacg
      541 gtaccgcggg cccgggatcc accggtcgcc accatggtgt ctaagggcga agagctgatc
      601 aaggaaaata tgcgtatgaa ggtggtcatg gaaggttcgg tcaacggcca ccaattcaaa
      661 tgcacaggtg aaggagaagg caatccgtac atgggaactc aaaccatgag gatcaaagtc
      721 atcgagggag gacccctgcc atttgccttt gacattcttg ccacgtcgtt catgtatggc
      781 agccgtactt ttatcaagta cccgaaaggc attcctgatt tctttaaaca gtcctttcct
      841 gagggtttta cttgggaaag agttacgaga tacgaagatg gtggagtcgt caccgtcatg
      901 caggacacca gccttgagga tggctgtctc gtttaccacg tccaagtcag aggggtaaac
      961 tttccctcca atggtcccgt gatgcagaag aagaccaagg gttgggagcc taatacagag
     1021 atgatgtatc cagcagatgg tggtctgagg ggatacactc atatggcact gaaagttgat
     1081 ggtggtggcc atctgtcttg ctctttcgta acaacttaca ggtcaaaaaa gaccgtcggg
     1141 aacatcaaga tgcccggtat ccatgccgtt gatcaccgcc tggaaaggtt agaggaaagt
     1201 gacaatgaaa tgttcgtagt acaacgcgaa cacgcagttg ccaagttcgc cgggcttggt
     1261 ggtgggatgg acgagctgta caagtaagcg gccgcgactc tagatcataa tcagccatac
     1321 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     1381 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     1441 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     1501 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     1561 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     1621 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     1681 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     1741 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     1801 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     1861 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     1921 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     1981 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2041 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2101 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     2161 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2221 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2281 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     2341 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     2401 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     2461 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     2521 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     2581 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     2641 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     2701 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     2761 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     2821 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     2881 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     2941 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3001 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     3061 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3121 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3181 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3241 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3301 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     3361 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3421 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3481 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     3541 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     3601 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     3661 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     3721 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     3781 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3841 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3901 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3961 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4021 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4081 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4141 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4201 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4261 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4321 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4381 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4441 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4501 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4561 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     4621 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     4681 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     4741 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     4801 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     4861 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     4921 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     4981 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     5041 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     5101 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     5161 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagccac
     5221 c
//
