LOCUS       sequence-316721-        7417 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Alpha Actinin, Excitation: 559, Emission: 600.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7417)
  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 7417)
  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-Alpha-Actinin-19
FEATURES             Location/Qualifiers
     source          1..7417
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        101..404
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        405..608
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     559..579
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3404..3413
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3410..4120
                     /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    4245..4366
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4282..4301)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4336..4355
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4373..4828)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4510..4531)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4722..4741
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4855..4959
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4961..5318
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4961..5152
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4984..5004)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5169..5304
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5353..6147
                     /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             5353..6147
                     /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(5407..5426)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6017..6036
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6335..6354)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6379..6426
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6755..7343
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7244..7263
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa
       61 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      121 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      181 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      241 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      301 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      361 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      421 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      481 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      541 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      601 gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggactcag atctcgagct
      661 caagcttcgc accatcatgg accattatga ttctcagcaa accaacgatt acatgcagcc
      721 agaagaggac tgggaccggg acctgctcct ggacccggcc tgggagaagc agcagagaaa
      781 gacattcacg gcatggtgta actcccacct ccggaaggcg gggacacaga tcgagaacat
      841 cgaagaggac ttccgggatg gcctgaagct catgctgctg ctggaggtca tctcaggtga
      901 acgcttggcc aagccagagc gaggcaagat gagagtgcac aagatctcca acgtcaacaa
      961 ggccctggat ttcatagcca gcaaaggcgt caaactggtg tccatcggag ccgaagaaat
     1021 cgtggatggg aatgtgaaga tgaccctggg catgatctgg accatcatcc tgcgctttgc
     1081 catccaggac atctccgtgg aagagacttc agccaaggaa gggctgctcc tgtggtgtca
     1141 gagaaagaca gccccttaca aaaatgtcaa catccagaac ttccacataa gctggaagga
     1201 tggcctcggc ttctgtgctt tgatccaccg acaccggccc gagctgattg actacgggaa
     1261 gctgcggaag gatgatccac tcacaaatct gaatacggct tttgacgtgg cagagaagta
     1321 cctggacatc cccaagatgc tggatgccga agacatcgtt ggaactgccc gaccggatga
     1381 gaaagccatc atgacttacg tgtctagctt ctaccacgcc ttctctggag cccagaaggc
     1441 ggagacagca gccaatcgca tctgcaaggt gttggccgtc aaccaggaga acgagcagct
     1501 tatggaagac tacgagaagc tggccagtga tctgttggag tggatccgcc gcacaatccc
     1561 gtggctggag aaccgggtgc ccgagaacac catgcatgcc atgcaacaga agctggagga
     1621 cttccgggac taccggcgcc tgcacaagcc gcccaaggtg caggagaagt gccagctgga
     1681 gatcaacttc aacacgctgc agaccaagct gcggctcagc aaccggcctg ccttcatgcc
     1741 ctctgagggc aggatggtct cggacatcaa caatgcctgg ggctgcctgg agcaggtgga
     1801 gaagggctat gaggagtggt tgctgaatga gatccggagg ctggagcgac tggaccacct
     1861 ggcagagaag ttccggcaga aggcctccat ccacgaggcc tggactgacg gcaaagaggc
     1921 catgctgcga cagaaggact atgagaccgc caccctctcg gagatcaagg ccttgctcaa
     1981 gaagcatgag gccttcgaga gtgacctggc tgcccaccag gaccgtgtgg agcagattgc
     2041 cgccatcgca caggagctca atgagctgga ctattatgac tcacccagtg tcaacgcccg
     2101 ttgccaaaag atctgtgacc agtgggacaa tctgggggcc ctaactcaga agcgaaggga
     2161 agctctggag cggaccgaga aactgctgga gaccattgac cagctgtact tggagtatgc
     2221 caagcgggct gcacccttca acaactggat ggagggggcc atggaggacc tgcaggacac
     2281 cttcattgtg cacaccattg aggagatcca gggactgacc acagcccatg agcagttcaa
     2341 ggccaccctc cctgatgccg acaaggagcg cctggccatc ctgggcatcc acaatgaggt
     2401 gtccaagatt gtccagacct accacgtcaa tatggcgggc accaacccct acacaaccat
     2461 cacgcctcag gagatcaatg gcaaatggga ccacgtgcgg cagctggtgc ctcggaggga
     2521 ccaagctctg acggaggagc atgcccgaca gcagcacaat gagaggctac gcaagcagtt
     2581 tggagcccag gccaatgtca tcgggccctg gatccagacc aagatggagg agatcgggag
     2641 gatctccatt gagatgcatg ggaccctgga ggaccagctc agccacctgc ggcagtatga
     2701 gaagagcatc gtcaactaca agccaaagat tgatcagctg gagggcgacc accagctcat
     2761 ccaggaggcg ctcatcttcg acaacaagca caccaactac accatggagc acatccgtgt
     2821 gggctgggag cagctgctca ccaccatcgc caggaccatc aatgaggtag agaaccagat
     2881 cctgacccgg gatgccaagg gcatcagcca ggagcagatg aatgagttcc gggcctcctt
     2941 caaccacttt gaccgggatc actccggcac actgggtccc gaggagttca aagcctgcct
     3001 catcagcttg ggttatgata ttggcaacga cccccaggga gaagcagaat ttgcccgcat
     3061 catgagcatt gtggacccca accgcctggg ggtagtgaca ttccaggcct tcattgactt
     3121 catgtcccgc gagacagccg acacagatac agcagaccaa gtcatggctt ccttcaagat
     3181 cctggctggg gacaagaact acattaccat ggacgagctg cgccgcgagc tgccacccga
     3241 ccaggctgag tactgcatcg cgcggatggc cccctacacc ggccccgact ccgtgccagg
     3301 tgctctggac tacatgtcct tctccacggc gctgtacggc gagagtgacc tcaagcttcg
     3361 aattctgcag tcgacggtac cgcgggcccg ggatccaccg gtcgccacca tggtgtctaa
     3421 gggcgaagag ctgatcaagg aaaatatgcg tatgaaggtg gtcatggaag gttcggtcaa
     3481 cggccaccaa ttcaaatgca caggtgaagg agaaggcaat ccgtacatgg gaactcaaac
     3541 catgaggatc aaagtcatcg agggaggacc cctgccattt gcctttgaca ttcttgccac
     3601 gtcgttcatg tatggcagcc gtacttttat caagtacccg aaaggcattc ctgatttctt
     3661 taaacagtcc tttcctgagg gttttacttg ggaaagagtt acgagatacg aagatggtgg
     3721 agtcgtcacc gtcatgcagg acaccagcct tgaggatggc tgtctcgttt accacgtcca
     3781 agtcagaggg gtaaactttc cctccaatgg tcccgtgatg cagaagaaga ccaagggttg
     3841 ggagcctaat acagagatga tgtatccagc agatggtggt ctgaggggat acactcatat
     3901 ggcactgaaa gttgatggtg gtggccatct gtcttgctct ttcgtaacaa cttacaggtc
     3961 aaaaaagacc gtcgggaaca tcaagatgcc cggtatccat gccgttgatc accgcctgga
     4021 aaggttagag gaaagtgaca atgaaatgtt cgtagtacaa cgcgaacacg cagttgccaa
     4081 gttcgccggg cttggtggtg ggatggacga gctgtacaag taagcggccg cgactctaga
     4141 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     4201 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     4261 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4321 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     4381 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     4441 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     4501 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     4561 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     4621 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4681 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4741 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4801 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4861 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4921 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     4981 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     5041 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     5101 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     5161 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5221 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     5281 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     5341 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5401 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5461 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5521 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5581 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5641 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5701 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5761 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5821 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5881 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5941 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     6001 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     6061 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     6121 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     6181 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     6241 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     6301 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     6361 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     6421 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     6481 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     6541 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     6601 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6661 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6721 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6781 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6841 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6901 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6961 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     7021 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7081 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     7141 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     7201 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     7261 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7321 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7381 tttgctggcc ttttgctcac atgttctttc ctgcgtt
//
