LOCUS       sequence-233874-        5347 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter for RNA nuclear localization.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5347)
  AUTHORS   Lubelsky Y, Ulitsky I
  TITLE     Sequences enriched in Alu repeats drive nuclear localization of long
            RNAs in human cells.
  JOURNAL   Nature. 2018 Mar 1;555(7694):107-111. doi: 10.1038/nature25757. Epub
            2018 Jan 24.
  PUBMED    29466324
REFERENCE   2  (bases 1 to 5347)
  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: 
            "10.1038/nature25757"; journalName: "Nature"; date: "2018-03-1- 1"; 
            volume: "555"; issue: "7694"; pages: "107-111"
COMMENT     Sequence Label: pAcGFP-MALAT1M
FEATURES             Location/Qualifiers
     source          1..5347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        54..357
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        358..561
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     512..532
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      600..609
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             606..1322
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    2128..2249
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2165..2184)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2219..2238
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2256..2711)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2393..2414)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2605..2624
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2738..2842
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2844..3201
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2844..3035
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2867..2887)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3052..3187
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3236..4030
                     /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             3236..4030
                     /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(3290..3309)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3900..3919
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4218..4237)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4262..4309
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4638..5226
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5127..5146
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
       61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg
      601 ccaccatggt gagcaagggc gccgagctgt tcaccggcat cgtgcccatc ctgatcgagc
      661 tgaatggcga tgtgaatggc cacaagttca gcgtgagcgg cgagggcgag ggcgatgcca
      721 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgcct gtgccctggc
      781 ccaccctggt gaccaccctg agctacggcg tgcagtgctt ctcacgctac cccgatcaca
      841 tgaagcagca cgacttcttc aagagcgcca tgcctgaggg ctacatccag gagcgcacca
      901 tcttcttcga ggatgacggc aactacaagt cgcgcgccga ggtgaagttc gagggcgata
      961 ccctggtgaa tcgcatcgag ctgaccggca ccgatttcaa ggaggatggc aacatcctgg
     1021 gcaataagat ggagtacaac tacaacgccc acaatgtgta catcatgacc gacaaggcca
     1081 agaatggcat caaggtgaac ttcaagatcc gccacaacat cgaggatggc agcgtgcagc
     1141 tggccgacca ctaccagcag aataccccca tcggcgatgg ccctgtgctg ctgcccgata
     1201 accactacct gtccacccag agcgccctgt ccaaggaccc caacgagaag cgcgatcaca
     1261 tgatctactt cggcttcgtg accgccgccg ccatcaccca cggcatggat gagctgtaca
     1321 agtccggact cagatcataa cctttagtct tttccagatg caaccttaaa atcagtgaca
     1381 agaaacattc caaacaagca acagtcttca agaaattaaa ctggcaagtg gaaatgttta
     1441 aacagttcag tgatctttag tgcattgttt atgtgtgggt ttctctctcc cctcccttgg
     1501 tcttaattct tacatgcagg aacactcagc agacacacgt atgcgaaggg ccagagaagc
     1561 cagacccagt aagaaaaaat agcctattta ctttaaataa accaaacatt ccgtttaaaa
     1621 tgtggggatt gggaaccact agttctttca gatggtattc ttcagactat agaaggagct
     1681 tccagttgaa ttcaccagtg gacaaaatga ggaaaacagg tgaacaagct ttttctgtat
     1741 ttacatacaa agtcagatca gttatgggac aatagtattg aatagatttc agctttatgc
     1801 tggagtaact ggcatgtgag caaactgtgt tggcgtgggg gtggaggggt gaggtgggcg
     1861 ctaagccttt ttttaagatt tttcaggtac ccctcactaa aggcaccgaa ggcttaaagt
     1921 aggacaacca tggagccttc ctgtggcagg agagacaaca aagcgttcaa ttctgcagtc
     1981 gacggtaccg cgggcccggg atccaccgga tctagataac tgatcataat cagccatacc
     2041 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     2101 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     2161 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2221 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt
     2281 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2341 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     2401 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2461 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     2521 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     2581 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     2641 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     2701 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2761 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2821 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     2881 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2941 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3001 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3061 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3121 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3181 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     3241 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3301 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3361 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     3421 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3481 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3541 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3601 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3661 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     3721 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     3781 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     3841 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     3901 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     3961 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4021 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4081 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4141 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4201 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     4261 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     4321 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     4381 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     4441 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     4501 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4561 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4621 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4681 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4741 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4801 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4861 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4921 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4981 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5041 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5101 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5161 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5221 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5281 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
     5341 tagttat
//
