LOCUS       sequence-412563-        5985 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Note- This plasmid contains Keratin 18. Localization: Cytokeratin, 
            Excitation: 549, Emission: 565.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5985)
  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 5985)
  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-Keratin-17
FEATURES             Location/Qualifiers
     source          1..5985
                     /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      1875..1884
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1881..2591
                     /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    2713..2834
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2750..2769)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2804..2823
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2841..3296)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2978..2999)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3190..3209
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3323..3427
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3429..3786
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3429..3620
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3452..3472)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3637..3772
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3821..4615
                     /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             3821..4615
                     /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(3875..3894)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4485..4504
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4803..4822)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4847..4894
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5223..5811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5712..5731
                     /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 gctagcacca
      541 tgagcttcac cactcgctcc accttctcca ccaactaccg gtccctgggc tctgtccagg
      601 cgcccagcta cggcgcccgg ccggtcagca gcgcggccag cgtctatgca ggcgctgggg
      661 gctctggttc ccggatctcc gtgtcccgct ccaccagctt caggggcggc atggggtccg
      721 ggggcctggc caccgggata gccgggggtc tggcaggaat gggaggcatc cagaacgaga
      781 aggagaccat gcaaagcctg aacgaccgcc tggcctctta cctggacaga gtgaggagcc
      841 tggagaccga gaaccggagg ctggagagca aaatccggga gcacttggag aagaagggac
      901 cccaggtcag agactggagc cattacttca agatcatcga ggacctgagg gctcagatct
      961 tcgcaaatac tgtggacaat gcccgcatcg ttctgcagat tgacaatgcc cgtcttgctg
     1021 ctgatgactt tagagtcaag tatgagacag agctggccat gcgccagtct gtggagaacg
     1081 acatccatgg gctccgcaag gtcattgatg acaccaatat cacacgactg cagctggaga
     1141 cagagatcga ggctctcaag gaggagctgc tcttcatgaa gaagaaccac gaagaggaag
     1201 taaaaggcct acaagcccag attgccagct ctgggttgac cgtggaggta gatgccccca
     1261 aatctcagga cctcgccaag atcatggcag acatccgggc ccaatatgac gagctggctc
     1321 ggaagaaccg agaggagcta gacaagtact ggtctcagca gattgaggag agcaccacag
     1381 tgctcaccac acagtctgct gaggttggag ctgctgagac gacgctcaca gagctgagac
     1441 gtacagtcca gtccttggag atcgacctgg actccatgag aaatctgaag gccagcttgg
     1501 agaacagcct gagggaggtg gaggcccgct acgccctaca gatggagcag ctcaacggga
     1561 tcctgctgca ccttgagtca gagctggcac agacccgggc agagggacag cgccaggccc
     1621 aggagtatga ggccctgctg aacatcaagg tcaagctgga ggctgagatc gccacctacc
     1681 gccgcctgct ggaagatggc gaggacttta atcttggtga tgccttggac agcagcaact
     1741 ccatgcaaac catccaaaag accaccaccc gccggatagt ggatggcaaa gtggtgtctg
     1801 agaccaatga caccaaagtt ctgaggcatc gaattctgca gtcgacggta ccgcgggccc
     1861 gggatccacc ggtcgccacc atggtgagca agggcgagga gaataacatg gccatcatca
     1921 aggagttcat gcgcttcaag gtgcgcatgg agggctccgt gaacggccac gagttcgaga
     1981 tcgagggcga gggcgagggc cgcccctacg agggctttca gaccgctaag ctgaaggtga
     2041 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcatttc acctacggct
     2101 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttcaagctg tccttccccg
     2161 agggcttcaa gtgggagcgc gtgatgaact acgaggacgg cggcgtggtg accgtgaccc
     2221 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
     2281 tcccctccga cggccccgtg atgcagaaga agaccatggg ctgggaggcc tcctccgagc
     2341 ggatgtaccc cgaggacggt gccctgaagg gcaagatcaa gatgaggctg aagctgaagg
     2401 acggcggcca ctacacctcc gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc
     2461 tgcccggcgc ctacatcgtc gacatcaagt tggacatcac ctcccacaac gaggactaca
     2521 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
     2581 tgtacaagta agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt
     2641 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     2701 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2761 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2821 atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     2881 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2941 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     3001 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     3061 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     3121 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     3181 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     3241 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     3301 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3361 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3421 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     3481 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3541 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3601 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3661 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3721 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3781 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3841 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3901 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3961 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     4021 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4081 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4141 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4201 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4261 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4321 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     4381 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4441 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4501 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4561 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4621 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4681 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4741 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     4801 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     4861 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     4921 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     4981 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     5041 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     5101 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5161 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5221 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5281 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5341 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     5401 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5461 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5521 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5581 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5641 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5701 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5761 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5821 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5881 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
     5941 atcaattacg gggtcattag ttcatagccc atatatggag ttccg
//
