LOCUS       sequence-243851-        4723 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 4723)
  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 4723)
  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-N1
FEATURES             Location/Qualifiers
     source          1..4723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..122
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12..31)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     66..85
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(244..954)
                     /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     complement(522..541)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     regulatory      951..960
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    962..1042
                     /label=MCS
                     /note="multiple cloning site"
     promoter        complement(1065..1268)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1094..1114)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1269..1572
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1747..2335)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1827..1846)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2664..2711
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2736..2755
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2943..3737)
                     /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             complement(2943..3737)
                     /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(3054..3073)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3664..3683
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3772..4129)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3786..3921
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3938..4129
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4086..4106
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4131..4235)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4262..4717
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4349..4368)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4559..4580
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
       61 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      121 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
      181 tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcta gagtcgcggc
      241 cgcttacttg tacagctcgt ccatgccgcc ggtggagtgg cggccctcgg cgcgttcgta
      301 ctgttccacg atggtgtagt cctcgttgtg ggaggtgatg tccaacttga tgtcgacgat
      361 gtaggcgccg ggcaactgca cgggcttctt ggccttgtag gtggtcttga cctcggaggt
      421 gtagtggccg ccgtccttca gcttcagcct gagcttgtcc tcgcccttca gggcgccgtc
      481 ctcggggtac atccgctcgg aggaggcctc catgcccatg gtcttcttct gcattacggg
      541 gccgtcggag gggaagttgg tgccgcgcag cttcaccttg tagatgaact cgccgtcctg
      601 cagggaggag tcctgagtca cggtcaccac gccgccgtcc tcgaagttca tcacgcgctc
      661 ccacttgaag ccctcgggga aggacagctt gaggtagtcg gggatgtcgg cggggtgctt
      721 cacgtaggcc ttggagccgt aggtgaactg aggggacaag atgtcccagg cgaagggcag
      781 ggggccaccc ttggtcacct tcagcttaac ggtctgaaag ccctcgtagg ggcggccctc
      841 gccctcgccc tcgatctcga actcgtggcc gttcacggag ccctccatgc gcaccttgaa
      901 gcgcatgaac tccttgatga tggccatgtt attctcctcg cccttgctca ccatggtggc
      961 gaccggtgga tcccgggccc gcggtaccgt cgactgcaga attcgaagct tgagctcgag
     1021 atctgagtcc ggtagcgcta gcggatctga cggttcacta aaccagctct gcttatatag
     1081 acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat
     1141 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg
     1201 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac
     1261 cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
     1321 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
     1381 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
     1441 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
     1501 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
     1561 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac
     1621 tattaataac taatgcatgg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1681 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1741 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1801 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1861 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1921 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1981 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2041 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2101 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2161 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2221 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2281 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2341 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2401 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2461 ttttaaatca atctaaagta tatatgagta acctgaggct atggcagggc ctgccgcccc
     2521 gacgttggct gcgagccctg ggccttcacc cgaacttggg gggtggggtg gggaaaagga
     2581 agaaacgcgg gcgtattggc cccaatgggg tctcggtggg gtatcgacag agtgccagcc
     2641 ctgggaccga accccgcgtt tatgaacaaa cgacccaaca ccgtgcgttt tattctgtct
     2701 ttttattgcc gtcatagcgc gggttccttc cggtattgtc tccttccgtg tttcagttag
     2761 cctcccccta gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca
     2821 gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc
     2881 gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc
     2941 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
     3001 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
     3061 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg
     3121 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
     3181 acgacgagat cctcgccgtc gggcatgctc gccttgagcc tggcgaacag ttcggctggc
     3241 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     3301 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     3361 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     3421 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     3481 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     3541 cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     3601 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     3661 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     3721 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcgat ctttgcaaaa
     3781 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     3841 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     3901 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     3961 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
     4021 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     4081 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcagg actcttcctt
     4141 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4201 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4261 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4321 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4381 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4441 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4501 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4561 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4621 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4681 aacgcgaatt ttaacaaaat attaacgctt acaatttacg cct
//
