LOCUS       sequence-111671-        5621 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses CNL-RhoB in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5621)
  AUTHORS   Takai A, Nakano M, Saito K, Haruno R, Watanabe TM, Ohyanagi T, Jin 
            T, Okada Y, Nagai T
  TITLE     Expanded palette of Nano-lanterns for real-time multicolor 
            luminescence imaging.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Apr 7;112(14):4352-6. doi: 
            10.1073/pnas.1418468112. Epub 2015 Mar 23.
  PUBMED    25831507
REFERENCE   2  (bases 1 to 5621)
  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: 
            "10.1073/pnas.1418468112"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2015-04-7- 7"; volume: "112"; issue: "14"; pages: "4352-6"
COMMENT     Sequence Label: pCNL-C1_RhoB
FEATURES             Location/Qualifiers
     source          1..5621
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     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      597..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(603..605,606..608,609..1121)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(648..669)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(909..928)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1256..1277
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2220..2285
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2409..2530
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2446..2465)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2500..2519
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2537..2992)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2674..2695)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2886..2905
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3019..3123
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3125..3482
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3125..3316
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3148..3168)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3333..3468
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3517..4311
                     /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             3517..4311
                     /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(3571..3590)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4181..4200
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4499..4518)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4543..4590
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4919..5507
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5408..5427
                     /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 gctagcgcca
      601 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
      661 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
      721 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
      781 ccctcgtgac caccctgtcc tggggcgtgc agtgcttcgc ccgctacccc gaccacatga
      841 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
      901 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
      961 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1021 acaagctgga gtacaactac tttagcgaca acgtctatat caccgccgac aagcagaaga
     1081 acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg
     1141 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1201 actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     1261 tcctgctgga gttcgtgacc gccgccgggg gtaccaaggt gtacgacccc gagcagagga
     1321 agaggatgat caccggcccc cagtggtggg ccaggtgcaa gcagatgaac gtgctggaca
     1381 gcttcatcaa ctactacgac agcgagaagc acgccgagaa cgccgtgatc ttcctgcacg
     1441 gcaacgccac tagcagctac ctgtggaggc acgtggtgcc ccacatcgag cccgtggcca
     1501 ggtgcatcat ccccgatctg atcggcatgg gcaagagcgg caagagcggc aacggcagct
     1561 acaggctgct ggaccactac aagtacctga ccgcctggtt cgagcttctg aacctgccca
     1621 agaagatcat cttcgtgggc cacgactggg gcgccgccct ggccttccac tacgcctacg
     1681 agcaccagga caggatcaag gccatcgtgc acatggagag cgtggtggac gtgatcgaga
     1741 gctgggacga gtggccagac atcgaggagg acatcgccct gatcaagagc gaggagggcg
     1801 agaagatggt gctggagaac aacttcttcg tggagaccgt gctgcccagc aagatcatga
     1861 gaaagctgga gcccgaggag ttcgccgcct acctggagcc cttcaaggag aagggcgagg
     1921 tgagaagacc caccctgagc tggcccagag agatccccct ggtgaagggc ggcaagcccg
     1981 acgtggtgca gatcgtgaga aactacaacg cctacctgag agccagcgac gacctgccca
     2041 agctgttcat cgagggcgac cccggcttct tcagcaacgc catcgtggag ggcgccaaga
     2101 agttccccaa caccgagttc gtgaaggtga agggcctgca cttcctccag gaggacgccc
     2161 ccgacgagat gggcaagtac atcaagagct tcgtggagag agtgctgaag aacgagcagt
     2221 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
     2281 gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac
     2341 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     2401 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2461 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2521 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     2581 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     2641 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     2701 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2761 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2821 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     2881 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     2941 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     3001 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3061 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3121 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3181 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3241 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3301 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3361 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3421 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3481 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3541 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3601 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3661 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     3721 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3781 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3841 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3901 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3961 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4021 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     4081 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4141 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4201 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4261 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4321 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4381 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4441 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     4501 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     4561 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     4621 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     4681 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     4741 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     4801 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4861 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4921 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4981 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5041 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5101 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5161 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5221 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5281 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5341 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5401 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5461 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5521 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5581 tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
