LOCUS       sequence-46731-e        7365 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 7365)
  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 7365)
  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: pActinin-LSSmOrange
FEATURES             Location/Qualifiers
     source          1..7365
                     /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      3364..3373
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3370..4080
                     /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     3783..3802
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4202..4323
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4239..4258)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4293..4312
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4330..4785)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4467..4488)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4679..4698
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4812..4916
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4918..5275
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4918..5109
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4941..4961)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5126..5261
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5310..6104
                     /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             5310..6104
                     /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(5364..5383)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5974..5993
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6292..6311)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6336..6383
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6712..7300
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7201..7220
                     /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 gctagcgcta
      601 ccggactcag atctcgagct caagcttcgc accatcatgg accattatga ttctcagcaa
      661 accaacgatt acatgcagcc agaagaggac tgggaccggg acctgctcct ggacccggcc
      721 tgggagaagc agcagagaaa gacattcacg gcatggtgta actcccacct ccggaaggcg
      781 gggacacaga tcgagaacat cgaagaggac ttccgggatg gcctgaagct catgctgctg
      841 ctggaggtca tctcaggtga acgcttggcc aagccagagc gaggcaagat gagagtgcac
      901 aagatctcca acgtcaacaa ggccctggat ttcatagcca gcaaaggcgt caaactggtg
      961 tccatcggag ccgaagaaat cgtggatggg aatgtgaaga tgaccctggg catgatctgg
     1021 accatcatcc tgcgctttgc catccaggac atctccgtgg aagagacttc agccaaggaa
     1081 gggctgctcc tgtggtgtca gagaaagaca gccccttaca aaaatgtcaa catccagaac
     1141 ttccacataa gctggaagga tggcctcggc ttctgtgctt tgatccaccg acaccggccc
     1201 gagctgattg actacgggaa gctgcggaag gatgatccac tcacaaatct gaatacggct
     1261 tttgacgtgg cagagaagta cctggacatc cccaagatgc tggatgccga agacatcgtt
     1321 ggaactgccc gaccggatga gaaagccatc atgacttacg tgtctagctt ctaccacgcc
     1381 ttctctggag cccagaaggc ggagacagca gccaatcgca tctgcaaggt gttggccgtc
     1441 aaccaggaga acgagcagct tatggaagac tacgagaagc tggccagtga tctgttggag
     1501 tggatccgcc gcacaatccc gtggctggag aaccgggtgc ccgagaacac catgcatgcc
     1561 atgcaacaga agctggagga cttccgggac taccggcgcc tgcacaagcc gcccaaggtg
     1621 caggagaagt gccagctgga gatcaacttc aacacgctgc agaccaagct gcggctcagc
     1681 aaccggcctg ccttcatgcc ctctgagggc aggatggtct cggacatcaa caatgcctgg
     1741 ggctgcctgg agcaggtgga gaagggctat gaggagtggt tgctgaatga gatccggagg
     1801 ctggagcgac tggaccacct ggcagagaag ttccggcaga aggcctccat ccacgaggcc
     1861 tggactgacg gcaaagaggc catgctgcga cagaaggact atgagaccgc caccctctcg
     1921 gagatcaagg ccttgctcaa gaagcatgag gccttcgaga gtgacctggc tgcccaccag
     1981 gaccgtgtgg agcagattgc cgccatcgca caggagctca atgagctgga ctattatgac
     2041 tcacccagtg tcaacgcccg ttgccaaaag atctgtgacc agtgggacaa tctgggggcc
     2101 ctaactcaga agcgaaggga agctctggag cggaccgaga aactgctgga gaccattgac
     2161 cagctgtact tggagtatgc caagcgggct gcacccttca acaactggat ggagggggcc
     2221 atggaggacc tgcaggacac cttcattgtg cacaccattg aggagatcca gggactgacc
     2281 acagcccatg agcagttcaa ggccaccctc cctgatgccg acaaggagcg cctggccatc
     2341 ctgggcatcc acaatgaggt gtccaagatt gtccagacct accacgtcaa tatggcgggc
     2401 accaacccct acacaaccat cacgcctcag gagatcaatg gcaaatggga ccacgtgcgg
     2461 cagctggtgc ctcggaggga ccaagctctg acggaggagc atgcccgaca gcagcacaat
     2521 gagaggctac gcaagcagtt tggagcccag gccaatgtca tcgggccctg gatccagacc
     2581 aagatggagg agatcgggag gatctccatt gagatgcatg ggaccctgga ggaccagctc
     2641 agccacctgc ggcagtatga gaagagcatc gtcaactaca agccaaagat tgatcagctg
     2701 gagggcgacc accagctcat ccaggaggcg ctcatcttcg acaacaagca caccaactac
     2761 accatggagc acatccgtgt gggctgggag cagctgctca ccaccatcgc caggaccatc
     2821 aatgaggtag agaaccagat cctgacccgg gatgccaagg gcatcagcca ggagcagatg
     2881 aatgagttcc gggcctcctt caaccacttt gaccgggatc actccggcac actgggtccc
     2941 gaggagttca aagcctgcct catcagcttg ggttatgata ttggcaacga cccccaggga
     3001 gaagcagaat ttgcccgcat catgagcatt gtggacccca accgcctggg ggtagtgaca
     3061 ttccaggcct tcattgactt catgtcccgc gagacagccg acacagatac agcagaccaa
     3121 gtcatggctt ccttcaagat cctggctggg gacaagaact acattaccat ggacgagctg
     3181 cgccgcgagc tgccacccga ccaggctgag tactgcatcg cgcggatggc cccctacacc
     3241 ggccccgact ccgtgccagg tgctctggac tacatgtcct tctccacggc gctgtacggc
     3301 gagagtgacc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccaccg
     3361 gtcgccacca tggtgagcaa gggcgaggag aataacatgg ccatcatcaa ggagttcatg
     3421 cgcttcaagg tgcgcatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     3481 ggcgagggcc gcccctacga gggctttcag accgttaagc tgaaggtgac caagggtggc
     3541 cccctgccct tcgcctggga catcttgtcc cctcagttca cctacggctc caaggcctac
     3601 gtgaagcacc ccgccgacat ccccgactac ctcaagctgt ccttccccga gggcttcaag
     3661 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgactca ggactcctcc
     3721 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
     3781 ggccccgtaa tgcagaagaa gaccatgggc atggaggcct cctccgagcg gatgtacccc
     3841 gaggacggcg ccctgaaggg cgaggacaag ctcaggctga agctgaagga cggcggccac
     3901 tacacctccg aggtcaagac cacctacaag gccaagaagc ccgtgcagtt gcccggcgcc
     3961 tacatcgtcg acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
     4021 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa
     4081 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     4141 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     4201 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4261 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4321 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     4381 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     4441 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     4501 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     4561 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     4621 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     4681 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     4741 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     4801 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4861 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     4921 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4981 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     5041 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5101 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     5161 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     5221 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     5281 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     5341 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     5401 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5461 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     5521 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5581 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5641 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5701 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5761 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5821 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5881 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5941 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     6001 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     6061 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     6121 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     6181 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     6241 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     6301 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     6361 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     6421 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     6481 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     6541 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     6601 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6661 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6721 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6781 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6841 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6901 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6961 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     7021 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7081 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     7141 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7201 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7261 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7321 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctt
//
