LOCUS       sequence-46729-e        5985 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5985)
  AUTHORS   Shcherbakova DM, Hink MA, Joosen L, Gadella TW, Verkhusha VV
  TITLE     An Orange Fluorescent Protein with a Large Stokes Shift for 
            Single-Excitation Multicolor FCCS and FRET Imaging.
  JOURNAL   J Am Chem Soc. 2012 May 9;134(18):7913-23. doi: 10.1021/ja3018972
  PUBMED    22486524
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"; journalName: 
            "J Am Chem Soc"; date: "2012-05-9- 9"; volume: "134"; issue: "18"; 
            pages: "7913-23"
COMMENT     Sequence Label: pKeratin-LSSmOrange
FEATURES             Location/Qualifiers
     source          1..5985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1935..1944
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1941..2651
                     /codon_start=1
                     /product="monomeric orange fluorescent protein with a large
                     Stokes shift (Shcherbakova et al., 2012)"
                     /label=LSSmOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTVKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGMEASSERMYPEDGALK
                     GEDKLRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     2354..2373
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2773..2894
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2810..2829)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2864..2883
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2901..3356)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3038..3059)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3250..3269
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3383..3487
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3489..3846
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3489..3680
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3512..3532)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3697..3832
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3881..4675
                     /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             3881..4675
                     /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(3935..3954)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4545..4564
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4863..4882)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4907..4954
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5283..5871
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5772..5791
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcacca
      601 tgagcttcac cactcgctcc accttctcca ccaactaccg gtccctgggc tctgtccagg
      661 cgcccagcta cggcgcccgg ccggtcagca gcgcggccag cgtctatgca ggcgctgggg
      721 gctctggttc ccggatctcc gtgtcccgct ccaccagctt caggggcggc atggggtccg
      781 ggggcctggc caccgggata gccgggggtc tggcaggaat gggaggcatc cagaacgaga
      841 aggagaccat gcaaagcctg aacgaccgcc tggcctctta cctggacaga gtgaggagcc
      901 tggagaccga gaaccggagg ctggagagca aaatccggga gcacttggag aagaagggac
      961 cccaggtcag agactggagc cattacttca agatcatcga ggacctgagg gctcagatct
     1021 tcgcaaatac tgtggacaat gcccgcatcg ttctgcagat tgacaatgcc cgtcttgctg
     1081 ctgatgactt tagagtcaag tatgagacag agctggccat gcgccagtct gtggagaacg
     1141 acatccatgg gctccgcaag gtcattgatg acaccaatat cacacgactg cagctggaga
     1201 cagagatcga ggctctcaag gaggagctgc tcttcatgaa gaagaaccac gaagaggaag
     1261 taaaaggcct acaagcccag attgccagct ctgggttgac cgtggaggta gatgccccca
     1321 aatctcagga cctcgccaag atcatggcag acatccgggc ccaatatgac gagctggctc
     1381 ggaagaaccg agaggagcta gacaagtact ggtctcagca gattgaggag agcaccacag
     1441 tggtcaccac acagtctgct gaggttggag ctgctgagac gacgctcaca gagctgagac
     1501 gtacagtcca gtccttggag atcgacctgg actccatgag aaatctgaag gccagcttgg
     1561 agaacagcct gagggaggtg gaggcccgct acgccctaca gatggagcag ctcaacggga
     1621 tcctgctgca ccttgagtca gagctggcac agacccgggc agagggacag cgccaggccc
     1681 aggagtatga ggccctgctg aacatcaagg tcaagctgga ggctgagatc gccacctacc
     1741 gccgcctgct ggaagatggc gaggacttta atcttggtga tgccttggac agcagcaact
     1801 ccatgcaaac catccaaaag accaccaccc gccggatagt ggatggcaaa gtggtgtctg
     1861 agaccaatga caccaaagtt ctgaggcatc gaattctgca gtcgacggta ccgcgggccc
     1921 gggatccacc ggtcgccacc atggtgagca agggcgagga gaataacatg gccatcatca
     1981 aggagttcat gcgcttcaag gtgcgcatgg agggctccgt gaacggccac gagttcgaga
     2041 tcgagggcga gggcgagggc cgcccctacg agggctttca gaccgttaag ctgaaggtga
     2101 ccaagggtgg ccccctgccc ttcgcctggg acatcttgtc ccctcagttc acctacggct
     2161 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cctcaagctg tccttccccg
     2221 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgactc
     2281 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
     2341 tcccctccga cggccccgta atgcagaaga agaccatggg catggaggcc tcctccgagc
     2401 ggatgtaccc cgaggacggc gccctgaagg gcgaggacaa gctcaggctg aagctgaagg
     2461 acggcggcca ctacacctcc gaggtcaaga ccacctacaa ggccaagaag cccgtgcagt
     2521 tgcccggcgc ctacatcgtc gacatcaagt tggacatcac ctcccacaac gaggactaca
     2581 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
     2641 tgtacaagta agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt
     2701 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     2761 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2821 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2881 atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     2941 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     3001 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     3061 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     3121 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     3181 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     3241 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     3301 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     3361 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3421 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3481 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     3541 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3601 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3661 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3721 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3781 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3841 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3901 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3961 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4021 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     4081 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4141 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4201 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4261 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4321 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4381 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     4441 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4501 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4561 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4621 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4681 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4741 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4801 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     4861 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     4921 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     4981 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     5041 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     5101 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     5161 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5221 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5281 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5341 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5401 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     5461 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5521 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5581 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5641 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5701 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5761 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5821 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5881 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5941 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
