LOCUS       sequence-160207-        6536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter of mouse wildtype Nanog 3'UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6536)
  AUTHORS   Choi YJ, Lin CP, Ho JJ, He X, Okada N, Bu P, Zhong Y, Kim SY, 
            Bennett MJ, Chen C, Ozturk A, Hicks GG, Hannon GJ, He L
  TITLE     miR-34 miRNAs provide a barrier for somatic cell reprogramming.
  JOURNAL   Nat Cell Biol. 2011 Oct 23;13(11):1353-60. doi: 10.1038/ncb2366.
  PUBMED    22020437
REFERENCE   2  (bases 1 to 6536)
  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: 
            "Nat Cell Biol"; date: "2011-10-23- 23"; volume: "13"; issue: "11"; 
            pages: "1353-60"
COMMENT     Sequence Label: Luc-Nanog-3'UTR
FEATURES             Location/Qualifiers
     source          1..6536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    238..359
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(275..294)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     329..348
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    401..425
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1342..1366)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(1376..3028)
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     2944..2962
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     regulatory      3025..3034
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3064..3815)
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     misc_feature    3865..3956
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(3897..3916)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    3970..4018
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4149..4604)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4286..4307)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4498..4517
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4631..4735
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4736..4804,4805..5596)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4954..4973)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5767..6355
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6256..6275
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6509..6526
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
       61 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
      121 atcggtcgac ggatccttat cgattttacc acatttgtag aggttttact tgctttaaaa
      181 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
      241 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
      301 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
      361 catgtctgct cgaagcggcc ggccgccccg actctagatc accactttgt acaagaaagc
      421 tgggtcggcg cgcccaccct ttcaggaggc aaagataagt gggcagatct ctgagttcaa
      481 ggccaggctg gtctgcagag ctagttcaaa gacaattaga gctatgcaga gaaaccctgt
      541 cttgataaac aaagagcaaa taataattaa aaaaataaac caggggctgg tgagatggct
      601 cagtggataa gagcacccga ctgctcttcc gaaggtcagg agttcaaatc ccagcaacca
      661 catggtggct cacaaccata cgtaacaaga tctgacgccc tcttctggag tgtctgaaga
      721 cagctacagt gtacttacat ataatgaata aataaatctt taaaaaaaat atgaaaatat
      781 ttggaagaag gaaggaacct ggctttgccc tgactttaag cccagatgtt gcgtaagtct
      841 catatttcac ctggtggagt cacagagtag ttcaggaata attccaaggc ttgtggggtg
      901 ctaaaatgcg catggctttc cctagtggct tccaaattca cctccaaatc actggcagag
      961 aagttttgct gcaactgtac gtaaggctgc agaaagtcct ccccgaagtt atggagcgga
     1021 gcagcattcc aaggctggcc gttccaggac tgagcggtcc aggcctggct gctccaagtt
     1081 gggttggtcc aggtctggtt gttccaagtt gggttggtcc aggtctggct gctccaagtt
     1141 gggttggtcc aagtctggct gccccacatg gaaaggcttc cagatgcgtt caccagatag
     1201 ccctggggat agctgcaatg gatgctggga tactccactg gtgctgagcc cttctgaatc
     1261 agaccattgc tagtcttcaa ccactggttt ttctgccacc gcttgcactt atcctttggt
     1321 tggtgaaggg ggcggccgcg gagcctgctt ttttgtacaa acttgtgatc tagaattaca
     1381 cggcgatctt tccgcccttc ttggccttta tgaggatctc tctgattttt cttgcgtcga
     1441 gttttccggt aagacctttc ggtacttcgt ccacaaacac aactcctccg cgcaactttt
     1501 tcgcggttgt tacttgactg gcgacgtaat ccacgatctc tttttccgtc atcgtctttc
     1561 cgtgctccaa aacaacaacg gcggcgggaa gttcaccggc gtcatcgtcg ggaagacctg
     1621 cgacacctgc gtcgaagatg ttggggtgtt ggagcaagat ggattccaat tcagcgggag
     1681 ccacctgata gcctttgtac ttaatcagag acttcaggcg gtcaacgatg aagaagtgtt
     1741 cgtcttcgtc ccagtaagct atgtctccag aatgtagcca tccatccttg tcaatcaagg
     1801 cgttggtcgc ttccggattg tttacataac cggacataat cataggacct ctcacacaca
     1861 gttcgcctct ttgattaacg cccagcgttt tcccggtatc cagatccaca accttcgctt
     1921 caaaaaatgg aacaacttta ccgaccgcgc ccggtttatc atccccctcg ggtgtaatca
     1981 gaatagctga tgtagtctca gtgagcccat atccttgcct gatacctggc agatggaacc
     2041 tcttggcaac cgcttccccg acttccttag agaggggagc gccaccagaa gcaatttcgt
     2101 gtaaattaga taaatcgtat ttgtcaatca gagtgctttt ggcgaagaag gagaataggg
     2161 ttggcaccag cagcgcactt tgaatcttgt aatcctgaag gctcctcaga aacagctctt
     2221 cttcaaatct atacattaag acgactcgaa atccacatat caaatatccg agtgtagtaa
     2281 acattccaaa accgtgatgg aatggaacaa cacttaaaat cgcagtatcc ggaatgattt
     2341 gattgccaaa aataggatct ctggcatgcg agaatctcac gcaggcagtt ctatgaggca
     2401 gagcgacacc tttaggcaga ccagtagatc cagaggagtt catgatcagt gcaattgtct
     2461 tgtccctatc gaaggactct ggcacaaaat cgtattcatt aaaaccggga ggtagatgag
     2521 atgtgacgaa cgtgtacatc gactgaaatc cctggtaatc cgttttagaa tccatgataa
     2581 taattttttg gatgattggg agcttttttt gcacgttcaa aattttttgc aacccctttt
     2641 tggaaacgaa caccacggta ggctgcgaaa tgcccatact gttgagcaat tcacgttcat
     2701 tataaatgtc gttcgcgggc gcaactgcaa ctccgataaa taacgcgccc aacaccggca
     2761 taaagaattg aagagagttt tcactgcata cgacgattct gtgatttgta ttcagcccat
     2821 atcgtttcat agcttctgcc aaccgaacgg acatttcgaa gtactcagcg taagtgatgt
     2881 ccacctcgat atgtgcatct gtaaaagcaa ttgttccagg aaccagggcg tatctcttca
     2941 tagccttatg cagttgctct ccagcggttc catcttccag cggatagaat ggcgccgggc
     3001 ctttctttat gtttttggcg tcttccatgg tggctttacc aacagtaccg gaatgccaag
     3061 cttttaagcg ggtcgctgca gggtcgctcg gtgttcgagg ccacacgcgt caccttaata
     3121 tgcgaagtgg acctgggacc gcgccgcccc gactgcatct gcgtgttcga attcgccaat
     3181 gacaagacgc tgggcggggt ttgtgtcatc atagaactaa agacatgcaa atatatttct
     3241 tccggggaca ccgccagcaa acgcgagcaa cgggccacgg ggatgaagca gctgcgccac
     3301 tccctgaagc tcctgcagtc cctcgcgcct ccgggtgaca agatagtgta cctgtgcccc
     3361 gtcctggtgt ttgtcgccca acggacgctc cgcgtcagcc gcgtgacccg gctcgtcccg
     3421 cagaaggtct ccggtaatat caccgcagtc gtgcggatgc tccagagcct gtccacgtat
     3481 acggtcccca tggagcctag gacccagcga gcccgtcgcc gccgcggcgg cgccgcccgg
     3541 gggtctgcga gcagaccgaa aaggtcacac tctggggcgc gcgacccgcc cgagtcagcg
     3601 gcccgccagt taccacccgc cgaccaaacc cccgcctcca cggagggcgg gggggtgctt
     3661 aagaggatcg cggcgctctt ctgcgtgccc gtggccacca agaccaaacc ccgagccgcc
     3721 tccgaatgag agtgtttcgt tccttccccc tccccccgcg tcagacaaac cctaaccacc
     3781 gcttaagcgg cccccgcgag gtccgaagac tcatttagat ctcgagcccg ggctagcacg
     3841 cgtaagagct cggtacctat cgatagagaa atgttctggc acctgcactt gcactgggga
     3901 cagcctattt tgctagtttg ttttgtttcg ttttgttttg atggagagcg tatgttagta
     3961 ctatcgattc acacaaaaaa ccaacacaca gatgtaatga aaataaagat attttattgc
     4021 ggccgctcca agtacctccc gtaccttaat attacttact tatcatggta gcttgggctg
     4081 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     4141 cgaatggcaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     4201 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     4261 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     4321 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     4381 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     4441 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     4501 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     4561 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     4621 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4681 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4741 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4801 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4861 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4921 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4981 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5041 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5101 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5161 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5221 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5281 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5341 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5401 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5461 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5521 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5581 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5641 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5701 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5761 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5821 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5881 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5941 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6001 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6061 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6121 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6181 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6241 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6301 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6361 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6421 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     6481 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
