LOCUS       sequence-174027-        6038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Gap Junctions, Excitation: 559, Emission: 600.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6038)
  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 6038)
  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-Cx43-7
FEATURES             Location/Qualifiers
     source          1..6038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1162..1171
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1168..1878
                     /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    2003..2124
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2040..2059)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2094..2113
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2131..2586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2268..2289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2480..2499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2613..2717
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2719..3076
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2719..2910
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2742..2762)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2927..3062
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3111..3905
                     /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             3111..3905
                     /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(3165..3184)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3775..3794
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4093..4112)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4137..4184
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4513..5101
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5002..5021
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5276..5579
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5580..5783
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5734..5754
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 atgggtgact ggagtgcctt ggggaagctt ctggacaagg tccaagccta ctccaccgct
       61 ggagggaagg tgtggctgtc agtgctcttc atattcagaa tcctgctcct ggggacagct
      121 gttgagtcag cttggggtga tgaacagtct gcctttcgct gtaacactca acaacctggc
      181 tgcgaaaacg tctgctatga caagtccttc cccatctctc acgtgcgctt ctgggtcctt
      241 cagatcatat tcgtgtctgt gcccacactc ctgtacttgg cccatgtgtt ctatgtgatg
      301 aggaaggaag agaagctaaa caagaaagaa gaggagctca aagtggccca gactgacggg
      361 gtcaacgtgg agatgcacct gaagcagatt gaaatcaaga agttcaagta cgggattgaa
      421 gagcacggca aggtgaaaat gaggggcggc ttgctgagaa cctacatcat cagcatcctc
      481 ttcaagtctg tcttcgaggt ggccttcctg ctcatccagt ggtacatcta tgggttcagc
      541 ttgagcgcgg tctacacctg caagagagat ccctgccccc accaggtaga ctgcttcctc
      601 tcacgtccca cggagaaaac catcttcatc atcttcatgc tggtggtgtc cttggtgtct
      661 ctcgctttga acatcattga gctcttctac gtcttcttca agggcgttaa ggatcgcgtg
      721 aagggaagaa gcgatcctta ccacgccacc actggcccac tgagcccatc aaaagactgc
      781 ggatctccaa aatacgccta cttcaatggc tgctcctcac caacggctcc actctcgcct
      841 atgtctcctc ctgggtacaa gctggttact ggtgacagaa acaattcctc gtgccgcaat
      901 tacaacaagc aagctagcga gcaaaactgg gcgaactaca gcgcagagca aaatcgcatg
      961 gggcaggccg gaagcaccat ctccaactcg cacgcccagc cgttcgattt ccccgacgac
     1021 aaccagaatg ccaaaaaagt tgctgctgga catgaactcc agccattagc catcgtggac
     1081 caacgacctt ccagcagagc cagcagccgc gccagcagca ggcctcggcc tgatgacctg
     1141 gagattgcgg atccaccggt cgccaccatg gtgtctaagg gcgaagagct gatcaaggaa
     1201 aatatgcgta tgaaggtggt catggaaggt tcggtcaacg gccaccaatt caaatgcaca
     1261 ggtgaaggag aaggcaatcc gtacatggga actcaaacca tgaggatcaa agtcatcgag
     1321 ggaggacccc tgccatttgc ctttgacatt cttgccacgt cgttcatgta tggcagccgt
     1381 acttttatca agtacccgaa aggcattcct gatttcttta aacagtcctt tcctgagggt
     1441 tttacttggg aaagagttac gagatacgaa gatggtggag tcgtcaccgt catgcaggac
     1501 accagccttg aggatggctg tctcgtttac cacgtccaag tcagaggggt aaactttccc
     1561 tccaatggtc ccgtgatgca gaagaagacc aagggttggg agcctaatac agagatgatg
     1621 tatccagcag atggtggtct gaggggatac actcatatgg cactgaaagt tgatggtggt
     1681 ggccatctgt cttgctcttt cgtaacaact tacaggtcaa aaaagaccgt cgggaacatc
     1741 aagatgcccg gtatccatgc cgttgatcac cgcctggaaa ggttagagga aagtgacaat
     1801 gaaatgttcg tagtacaacg cgaacacgca gttgccaagt tcgccgggct tggtggtggg
     1861 atggacgagc tgtacaagta agcggccgcg actctagatc ataatcagcc ataccacatt
     1921 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     1981 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     2041 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2101 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     2161 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     2221 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     2281 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     2341 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     2401 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     2461 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     2521 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     2581 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2641 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2701 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     2761 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2821 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2881 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     2941 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3001 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3061 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3121 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3181 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3241 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     3301 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3361 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3421 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3481 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3541 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3601 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     3661 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3721 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3781 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     3841 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     3901 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     3961 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4021 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     4081 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     4141 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     4201 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     4261 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     4321 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     4381 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4441 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4501 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4561 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4621 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4681 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4741 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4801 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4861 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4921 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4981 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5041 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5101 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5161 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt
     5221 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     5281 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     5341 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     5401 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     5461 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     5521 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     5581 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     5641 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     5701 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     5761 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga
     5821 ctcagatctc gagctcaagc ttcgaattcg gcagcacttt tctttcattg gggggaaggc
     5881 gtgaggaaag taccaaacag cagcagactt ttaaacttta aacagacagg tctgagagcc
     5941 tgaactctca tttttccttt gacttcagcc tccaaggagt tccaccaact ttggcgcgcc
     6001 ggcttcactt tcattaagtg aaagagaggt gcccagac
//
