LOCUS       sequence-313814-        7414 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Alpha Actinin, Excitation: 549, Emission: 565.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7414)
  AUTHORS   Shaner NC, Lin MZ, McKeown MR, Steinbach PA, Hazelwood KL, Davidson 
            MW, Tsien RY
  TITLE     Improving the photostability of bright monomeric orange and red 
            fluorescent proteins.
  JOURNAL   Nat Methods. 2008 Jun;5(6):545-51. doi: 10.1038/nmeth.1209. Epub 
            2008 May 4.
  PUBMED    18454154
REFERENCE   2  (bases 1 to 7414)
  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"; doi: 
            "10.1038/nmeth.1209"; journalName: "Nat Methods"; date: "2008-06"; 
            volume: "5"; issue: "6"; pages: "545-51"
COMMENT     Sequence Label: mOrange2-Alpha-Actinin-19
FEATURES             Location/Qualifiers
     source          1..7414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        49..352
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        353..556
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     507..527
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3352..3361
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3358..4068
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     polyA_signal    4190..4311
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4227..4246)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4281..4300
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4318..4773)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4455..4476)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4667..4686
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4800..4904
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4906..5263
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4906..5097
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4929..4949)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5114..5249
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5298..6092
                     /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             5298..6092
                     /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(5352..5371)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5962..5981
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6280..6299)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6324..6371
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6700..7288
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7189..7208
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
       61 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      121 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      181 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      241 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      301 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      361 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      421 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      481 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      541 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggactcagat
      601 ctcgagctca agcttcgcac catcatggac cattatgatt ctcagcaaac caacgattac
      661 atgcagccag aagaggactg ggaccgggac ctgctcctgg acccggcctg ggagaagcag
      721 cagagaaaga cattcacggc atggtgtaac tcccacctcc ggaaggcggg gacacagatc
      781 gagaacatcg aagaggactt ccgggatggc ctgaagctca tgctgctgct ggaggtcatc
      841 tcaggtgaac gcttggccaa gccagagcga ggcaagatga gagtgcacaa gatctccaac
      901 gtcaacaagg ccctggattt catagccagc aaaggcgtca aactggtgtc catcggagcc
      961 gaagaaatcg tggatgggaa tgtgaagatg accctgggca tgatctggac catcatcctg
     1021 cgctttgcca tccaggacat ctccgtggaa gagacttcag ccaaggaagg gctgctcctg
     1081 tggtgtcaga gaaagacagc cccttacaaa aatgtcaaca tccagaactt ccacataagc
     1141 tggaaggatg gcctcggctt ctgtgctttg atccaccgac accggcccga gctgattgac
     1201 tacgggaagc tgcggaagga tgatccactc acaaatctga atacggcttt tgacgtggca
     1261 gagaagtacc tggacatccc caagatgctg gatgccgaag acatcgttgg aactgcccga
     1321 ccggatgaga aagccatcat gacttacgtg tctagcttct accacgcctt ctctggagcc
     1381 cagaaggcgg agacagcagc caatcgcatc tgcaaggtgt tggccgtcaa ccaggagaac
     1441 gagcagctta tggaagacta cgagaagctg gccagtgatc tgttggagtg gatccgccgc
     1501 acaatcccgt ggctggagaa ccgggtgccc gagaacacca tgcatgccat gcaacagaag
     1561 ctggaggact tccgggacta ccggcgcctg cacaagccgc ccaaggtgca ggagaagtgc
     1621 cagctggaga tcaacttcaa cacgctgcag accaagctgc ggctcagcaa ccggcctgcc
     1681 ttcatgccct ctgagggcag gatggtctcg gacatcaaca atgcctgggg ctgcctggag
     1741 caggtggaga agggctatga ggagtggttg ctgaatgaga tccggaggct ggagcgactg
     1801 gaccacctgg cagagaagtt ccggcagaag gcctccatcc acgaggcctg gactgacggc
     1861 aaagaggcca tgctgcgaca gaaggactat gagaccgcca ccctctcgga gatcaaggcc
     1921 ttgctcaaga agcatgaggc cttcgagagt gacctggctg cccaccagga ccgtgtggag
     1981 cagattgccg ccatcgcaca ggagctcaat gagctggact attatgactc acccagtgtc
     2041 aacgcccgtt gccaaaagat ctgtgaccag tgggacaatc tgggggccct aactcagaag
     2101 cgaagggaag ctctggagcg gaccgagaaa ctgctggaga ccattgacca gctgtacttg
     2161 gagtatgcca agcgggctgc acccttcaac aactggatgg agggggccat ggaggacctg
     2221 caggacacct tcattgtgca caccattgag gagatccagg gactgaccac agcccatgag
     2281 cagttcaagg ccaccctccc tgatgccgac aaggagcgcc tggccatcct gggcatccac
     2341 aatgaggtgt ccaagattgt ccagacctac cacgtcaata tggcgggcac caacccctac
     2401 acaaccatca cgcctcagga gatcaatggc aaatgggacc acgtgcggca gctggtgcct
     2461 cggagggacc aagctctgac ggaggagcat gcccgacagc agcacaatga gaggctacgc
     2521 aagcagtttg gagcccaggc caatgtcatc gggccctgga tccagaccaa gatggaggag
     2581 atcgggagga tctccattga gatgcatggg accctggagg accagctcag ccacctgcgg
     2641 cagtatgaga agagcatcgt caactacaag ccaaagattg atcagctgga gggcgaccac
     2701 cagctcatcc aggaggcgct catcttcgac aacaagcaca ccaactacac catggagcac
     2761 atccgtgtgg gctgggagca gctgctcacc accatcgcca ggaccatcaa tgaggtagag
     2821 aaccagatcc tgacccggga tgccaagggc atcagccagg agcagatgaa tgagttccgg
     2881 gcctccttca accactttga ccgggatcac tccggcacac tgggtcccga ggagttcaaa
     2941 gcctgcctca tcagcttggg ttatgatatt ggcaacgacc cccagggaga agcagaattt
     3001 gcccgcatca tgagcattgt ggaccccaac cgcctggggg tagtgacatt ccaggccttc
     3061 attgacttca tgtcccgcga gacagccgac acagatacag cagaccaagt catggcttcc
     3121 ttcaagatcc tggctgggga caagaactac attaccatgg acgagctgcg ccgcgagctg
     3181 ccacccgacc aggctgagta ctgcatcgcg cggatggccc cctacaccgg ccccgactcc
     3241 gtgccaggtg ctctggacta catgtccttc tccacggcgc tgtacggcga gagtgacctc
     3301 aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccggt cgccaccatg
     3361 gtgagcaagg gcgaggagaa taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     3421 cgcatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     3481 ccctacgagg gctttcagac cgctaagctg aaggtgacca agggtggccc cctgcccttc
     3541 gcctgggaca tcctgtcccc tcatttcacc tacggctcca aggcctacgt gaagcacccc
     3601 gccgacatcc ccgactactt caagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     3661 atgaactacg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     3721 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtgatg
     3781 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggtgcc
     3841 ctgaagggca agatcaagat gaggctgaag ctgaaggacg gcggccacta cacctccgag
     3901 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta catcgtcgac
     3961 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     4021 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaagc ggccgcgact
     4081 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     4141 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     4201 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4261 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     4321 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     4381 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     4441 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     4501 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     4561 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     4621 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     4681 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     4741 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     4801 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4861 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     4921 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4981 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     5041 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     5101 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     5161 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     5221 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     5281 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     5341 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     5401 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     5461 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     5521 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     5581 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     5641 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5701 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5761 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5821 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5881 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5941 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     6001 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     6061 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     6121 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     6181 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     6241 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     6301 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     6361 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     6421 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     6481 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     6541 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6601 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6661 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6721 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6781 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6841 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6901 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6961 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     7021 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     7081 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     7141 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     7201 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     7261 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     7321 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     7381 taaccgtatt accgccatgc attagttatt aata
//
