LOCUS       sequence-437549-        4859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To generate substrates to monitor Ku overloading and its impact on 
            transcription at the DNA ends vicinity..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4859)
  AUTHORS   Bossaert M, Moreno A, Peixoto A, Pillaire MJ, Chanut P, Frit P, 
            Calsou P, Loparo JJ, Britton S
  TITLE     Identification of the main barriers to Ku accumulation in chromatin.
  JOURNAL   bioRxiv [Preprint]. 2024 Jan 4:2024.01.03.574002. doi: 
            10.1101/2024.01.03.574002.
  PUBMED    38260538
REFERENCE   2  (bases 1 to 4859)
  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"; journalName: "bioRxiv 
            [Preprint]. 2024 Jan 4:2024.01.03.574002. doi: 
            10.1101/2024.01.03.574002."
COMMENT     Sequence Label: pUC18-LucFF
FEATURES             Location/Qualifiers
     source          1..4859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     30..52
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     44..61
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     45..61
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    101..325
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(326..1978)
                     /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     1894..1912
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     regulatory      1975..1984
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2023..2219)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        complement(2031..2241)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2037..2172
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2091..2110)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2265..2285
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(2304..2320)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2304..2320)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2317..2339)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2328..2344
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2352..2358,2359..2376,2377..2382))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2397..2418
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2535..2552)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2706..3294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2786..2805)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3465..4256,4257..4325))
                     /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     4088..4107
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4326..4430)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4498..4516
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4554..4576)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4676..4695
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
       61 tgccaagctt gcatgcctgc aggtcgactc tagaggatcc ccatagagcc caccgcatcc
      121 ccagcatgcc tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc caccccaccc
      181 cccagaatag aatgacacct actcagacaa tgcgatgcaa tttcctcatt ttattaggaa
      241 aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca
      301 gatggctggc aactagaagg cacagttaca cggcgatctt tccgcccttc ttggccttta
      361 tgaggatctc tctgattttt cttgcgtcga gttttccggt aagacctttc ggtacttcgt
      421 ccacaaacac aactcctccg cgcaactttt tcgcggttgt tacttgactg gcgacgtaat
      481 ccacgatctc tttttccgtc atcgtctttc cgtgctccaa aacaacaacg gcggcgggaa
      541 gttcaccggc gtcatcgtcg ggaagacctg cgacacctgc gtcgaagatg ttggggtgtt
      601 ggagcaagat ggattccaat tcagcgggag ccacctgata gcctttgtac ttaatcagag
      661 acttcaggcg gtcaacgatg aagaagtgtt cgtcttcgtc ccagtaagct atgtctccag
      721 aatgtagcca tccatccttg tcaatcaagg cgttggtcgc ttccggattg tttacataac
      781 cggacataat cataggacct ctcacacaca gttcgcctct ttgattaacg cccagcgttt
      841 tcccggtatc cagatccaca accttcgctt caaaaaatgg aacaacttta ccgaccgcgc
      901 ccggtttatc atccccctcg ggtgtaatca gaatagctga tgtagtctca gtgagcccat
      961 atccttgcct gatacctggc agatggaacc tcttggcaac cgcttccccg acttccttag
     1021 agaggggagc gccaccagaa gcaatttcgt gtaaattaga taaatcgtat ttgtcaatca
     1081 gagtgctttt ggcgaagaag gagaataggg ttggcaccag cagcgcactt tgaatcttgt
     1141 aatcctgaag gctcctcaga aacagctctt cttcaaatct atacattaag acgactcgaa
     1201 atccacatat caaatatccg agtgtagtaa acattccaaa accgtgatgg aatggaacaa
     1261 cacttaaaat cgcagtatcc ggaatgattt gattgccaaa aataggatct ctggcatgcg
     1321 agaatctcac gcaggcagtt ctatgaggca gagcgacacc tttaggcaga ccagtagatc
     1381 cagaggagtt catgatcagt gcaattgtct tgtccctatc gaaggactct ggcacaaaat
     1441 cgtattcatt aaaaccggga ggtagatgag atgtgacgaa cgtgtacatc gactgaaatc
     1501 cctggtaatc cgttttagaa tccatgataa taattttttg gatgattggg agcttttttt
     1561 gcacgttcaa aattttttgc aacccctttt tggaaacgaa caccacggta ggctgcgaaa
     1621 tgcccatact gttgagcaat tcacgttcat tataaatgtc gttcgcgggc gcaactgcaa
     1681 ctccgataaa taacgcgccc aacaccggca taaagaattg aagagagttt tcactgcata
     1741 cgacgattct gtgatttgta ttcagcccat atcgtttcat agcttctgcc aaccgaacgg
     1801 acatttcgaa gtactcagcg taagtgatgt ccacctcgat atgtgcatct gtaaaagcaa
     1861 ttgttccagg aaccagggcg tatctcttca tagccttatg cagttgctct ccagcggttc
     1921 catcttccag cggatagaat ggcgccgggc ctttctttat gtttttggcg tcttccatgg
     1981 tggcgctcga gcgaaaatgg atatacaagc tcccgggagc tttttgcaaa agcctaggcc
     2041 tccaaaaaag cctcctcact acttctggaa tagctcagag gcagaggcgg cctcggcctc
     2101 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
     2161 taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat
     2221 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
     2281 attccacagg aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc
     2341 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     2401 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2461 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2521 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2581 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2641 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2701 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2761 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2821 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2881 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2941 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3001 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3061 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3121 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3181 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3241 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3301 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3361 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3421 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3481 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3541 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3601 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3661 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3721 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3781 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3841 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3901 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3961 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4021 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4081 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4141 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4201 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4261 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4321 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4381 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4441 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     4501 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     4561 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     4621 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     4681 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     4741 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     4801 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaag
//
