LOCUS       Exported                6584 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses YNL-fibrillarin in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pYNL-C1_fibrillarin
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6584)
  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 6584)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6584
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             603..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
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     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"
     CDS             complement(3030..3041)
                     /codon_start=1
                     /gene="cspA"
                     /product="translation enhancing element for E. coli (Qing 
                     et al., 2004)"
                     /label=TEE
                     /translation="NHKV"
     polyA_signal    3372..3493
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3409..3428)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3463..3482
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3500..3955)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3637..3658)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3849..3868
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3982..4086
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4088..4445
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4111..4131)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4296..4431
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4358..4377
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4480..5274
                     /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(4534..4553)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5144..5163
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5462..5481)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5506..5553
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5882..6470
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6371..6390
                     /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 ctgatctgca ccaccggcaa gctgcccgtg ccctggccca
      781 ccctcgtgac caccctgggc tacggcctgc 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 aacagccaca acgtctatat caccgccgac aagcagaaga
     1081 acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg
     1141 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1201 actacctgag ctaccagtcc 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 attccatgaa gccaggattc agtccccgtg
     2281 ggggtggctt tggcggccga gggggctttg gtgaccgtgg tggtcgtgga ggccgagggg
     2341 gctttggcgg gggccgaggt cgaggcggag gctttagagg tcgtggacga ggaggaggtg
     2401 gaggcggcgg cggcggtgga ggaggaggaa gaggtggtgg aggcttccat tctggtggca
     2461 accggggtcg tggtcgggga ggaaaaagag gaaaccagtc ggggaagaat gtgatggtgg
     2521 agccgcatcg gcatgagggt gtcttcattt gtcgaggaaa ggaagatgca ctggtcacca
     2581 agaacctggt ccctggggaa tcagtttatg gagagaagag agtctcgatt tcggaaggag
     2641 atgacaaaat tgagtaccga gcctggaacc ccttccgctc caagctagca gcagcaatcc
     2701 tgggtggtgt ggaccagatc cacatcaaac cgggggctaa ggttctctac ctcggggctg
     2761 cctcgggcac cacggtctcc catgtctctg acatcgttgg tccggatggt ctagtctatg
     2821 cagtcgagtt ctcccaccgc tctggccgtg acctcattaa cttggccaag aagaggacca
     2881 acatcattcc tgtgatcgag gatgctcgac acccacacaa ataccgcatg ctcatcgcaa
     2941 tggtggatgt gatctttgct gatgtggccc agccagacca gacccggatt gtggccctga
     3001 atgcccacac cttcctgcgt aatggaggac actttgtgat ttccattaag gccaactgca
     3061 ttgactccac agcctcagcc gaggccgtgt ttgcctccga agtgaaaaag atgcaacagg
     3121 agaacatgaa gccgcaggag cagttgaccc ttgagccata tgaaagagac catgccgtgg
     3181 tcgtgggagt gtacaggcca ccccccaagg tgaagaactg agtcgacggt accgcgggcc
     3241 cgggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt
     3301 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     3361 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3421 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3481 tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     3541 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3601 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     3661 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     3721 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     3781 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     3841 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     3901 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     3961 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4021 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4081 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4141 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4201 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4261 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4321 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4381 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4441 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4501 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4561 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4621 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     4681 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4741 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4801 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4861 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4921 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4981 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     5041 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5101 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5161 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5221 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5281 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5341 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5401 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     5461 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     5521 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     5581 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     5641 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     5701 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     5761 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5821 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5881 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5941 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6001 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     6061 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6121 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6181 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6241 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6301 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6361 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6421 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6481 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6541 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
