LOCUS       sequence-46733-e        5806 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 5806)
  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 5806)
  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: pLSSmOrange-beta-Actin
FEATURES             Location/Qualifiers
     source          1..5806
                     /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      601..610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             607..1314
                     /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     1020..1039
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2594..2715
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2631..2650)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2685..2704
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2722..3177)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2859..2880)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3071..3090
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3204..3308
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3310..3667
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3310..3501
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3333..3353)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3518..3653
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3702..4496
                     /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             3702..4496
                     /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(3756..3775)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4366..4385
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4684..4703)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4728..4775
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5104..5692
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5593..5612
                     /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 gctagcggtc
      601 gccaccatgg tgagcaaggg cgaggagaat aacatggcca tcatcaagga gttcatgcgc
      661 ttcaaggtgc gcatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
      721 gagggccgcc cctacgaggg ctttcagacc gttaagctga aggtgaccaa gggtggcccc
      781 ctgcccttcg cctgggacat cttgtcccct cagttcacct acggctccaa ggcctacgtg
      841 aagcaccccg ccgacatccc cgactacctc aagctgtcct tccccgaggg cttcaagtgg
      901 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgactcagga ctcctccctg
      961 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     1021 cccgtaatgc agaagaagac catgggcatg gaggcctcct ccgagcggat gtaccccgag
     1081 gacggcgccc tgaagggcga ggacaagctc aggctgaagc tgaaggacgg cggccactac
     1141 acctccgagg tcaagaccac ctacaaggcc aagaagcccg tgcagttgcc cggcgcctac
     1201 atcgtcgaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     1261 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagttccgg
     1321 actcagatct cgagctatgg atgatgatat cgccgcgctc gtcgtcgaca acggctccgg
     1381 catgtgcaag gccggcttcg cgggcgacga tgccccccgg gccgtcttcc cctccatcgt
     1441 ggggcgcccc aggcaccagg gcgtgatggt gggcatgggt cagaaggatt cctatgtggg
     1501 cgacgaggcc cagagcaaga gaggcatcct caccctgaag taccccatcg agcacggcat
     1561 cgtcaccaac tgggacgaca tggagaaaat ctggcaccac accttctaca atgagctgcg
     1621 tgtggctccc gaggagcacc ccgtgctgct gaccgaggcc cccctgaacc ccaaggccaa
     1681 ccgcgagaag atgacccaga tcatgtttga gaccttcaac accccagcca tgtacgttgc
     1741 tatccaggct gtgctatccc tgtacgcctc tggccgtacc actggcatcg tgatggactc
     1801 cggtgacggg gtcacccaca ctgtgcccat ctacgagggg tatgccctcc cccatgccat
     1861 cctgcgtctg gacctggctg gccgggacct gactgactac ctcatgaaga tcctcaccga
     1921 gcgcggctac agcttcacca ccacggccga gcgggaaatc gtgcgtgaca ttaaggagaa
     1981 gctgtgctac gtcgccctgg acttcgagca agagatggcc acggctgctt ccagctcctc
     2041 cctggagaag agctacgagc tgcctgacgg ccaggtcatc accattggca atgagcggtt
     2101 ccgctgccct gaggcactct tccagccttc cttcctgggc atggagtcct gtggcatcca
     2161 cgaaactacc ttcaactcca tcatgaagtg tgacgtggac atccgcaaag acctgtacgc
     2221 caacacagtg ctgtctggcg gcaccaccat gtaccctggc attgccgaca ggatgcagaa
     2281 ggagatcact gccctggcac ccagcacaat gaagatcaag atcattgctc ctcctgagcg
     2341 caagtactcc gtgtggatcg gcggctccat cctggcctcg ctgtccacct tccagcagat
     2401 gtggatcagc aagcaggagt atgacgagtc cggcccctcc atcgtccacc gcaaatgctt
     2461 ctagggatcc accggatcta gataactgat cataatcagc cataccacat ttgtagaggt
     2521 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     2581 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2641 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     2701 catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2761 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2821 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     2881 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     2941 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     3001 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     3061 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3121 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     3181 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3241 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3301 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     3361 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3421 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3481 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3541 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3601 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3661 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3721 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3781 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3841 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     3901 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3961 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4021 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4081 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4141 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4201 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     4261 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4321 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4381 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4441 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4501 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4561 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4621 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     4681 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     4741 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     4801 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     4861 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     4921 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     4981 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5041 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5101 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5161 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5221 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     5281 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5341 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5401 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5461 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5521 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5581 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5641 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5701 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5761 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
