LOCUS       sequence-331146-        7071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of CSL R192H.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7071)
  AUTHORS   Bottoni G, Katarkar A, Tassone B, Ghosh S, Clocchiatti A, Goruppi S,
            Bordignon P, Jafari P, Tordini F, Lunardi T, Hoetzenecker W, Neel V,
            Lingner J, Paolo Dotto G
  TITLE     CSL controls telomere maintenance and genome stability in human 
            dermal fibroblasts.
  JOURNAL   Nat Commun. 2019 Aug 29;10(1):3884. doi: 10.1038/s41467-019-11785-7.
  PUBMED    31467287
REFERENCE   2  (bases 1 to 7071)
  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: "Nat 
            Commun."; date: "2019-08-29"; volume: "10(1):3884. doi"; pages: " 
            10.1038/s41467-019-11785-7"
COMMENT     Sequence Label: pcDNA3. Flag-CSL R192H
FEATURES             Location/Qualifiers
     source          1..7071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        155..534
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        535..738
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     689..709
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     783..802
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        783..801
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             824..847
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2544..2562)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2545..2562)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2582..2599)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2588..2812
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2858..3286
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2945..2964)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3155..3176
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3295..3315)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3300..3629
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3480..3615
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3542..3561
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3696..4490
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3750..3769)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4360..4379
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4664..4785
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4701..4720)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4755..4774
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4834..4850)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4834..4850)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4847..4869)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4858..4874
                     /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(4882..4888,4889..4906,4907..4912))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4927..4948
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5065..5082)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5236..5824)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5316..5335)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5995..6786,6787..6855))
                     /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     6618..6637
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6856..6960)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7035..7054)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
       61 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
      121 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
      181 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      241 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      301 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      361 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      421 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      481 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      541 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      601 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      661 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      721 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
      781 attaatacga ctcactatag ggagacccaa gcttggtacc atggactaca aagacgatga
      841 cgataaagga attctgcaga tgcctgttgt gacagggaaa tttggtgagc ggcctccacc
      901 taaacgactt actagggaag ctatgcgaaa ttatttaaaa gagcgagggg atcaaacagt
      961 acttattctt catgcaaaag ttgcacagaa gtcatatgga aatgaaaaaa ggtttttttg
     1021 cccacctcct tgtgtatatc ttatgggcag tggatggaag aaaaaaaaag aacaaatgga
     1081 acgcgatggt tgttctgaac aagagtctca accgtgtgca tttattggga taggaaatag
     1141 tgaccaagaa atgcagcagc taaacttgga aggaaagaac tattgcacag ccaaaacatt
     1201 gtatatatct gactcagaca agcgaaagca cttcatgttg tctgtaaaga tgttctatgg
     1261 caacagtgat gacattggtg tgttcctcag caagcggata aaagtcatct ccaaaccttc
     1321 caaaaagaag cagtcattga aaaatgctga cttatgcatt gcctcaggaa caaaggtggc
     1381 tctgtttaat cacctacgat cccagacagt tagtaccaga tacttgcatg tagaaggagg
     1441 taattttcat gccagttcac agcagtgggg agcctttttt attcatctct tggatgatga
     1501 tgaatcagaa ggagaagaat tcacagtccg agatggctac atccattatg gacaaacagt
     1561 caaacttgtg tgctcagtta ctggcatggc actcccaaga ttgataatta ggaaagttga
     1621 taagcagacc gcattattgg atgcagatga tcctgtgtca caactccata aatgtgcatt
     1681 ttaccttaag gatacagaaa gaatgtattt gtgcctttct caagaaagaa taattcaatt
     1741 tcaggccact ccatgtccaa aagaaccaaa taaagagatg ataaatgatg gcgcttcctg
     1801 gacaatcatt agcacagata aggcagagta tacattttat gagggaatgg gccctgtcct
     1861 tgccccagtc actcctgtgc ctgtggtaga gagccttcag ttgaatggcg gtggggacgt
     1921 agcaatgctt gaacttacag gacagaattt cactccaaat ttacgagtgt ggtttgggga
     1981 tgtagaagct gaaactatgt acaggtgtgg agagagtatg ctctgtgtcg tcccagacat
     2041 ttctgcattc cgagaaggtt ggagatgggt ccggcaacca gtccaggttc cagtaacttt
     2101 ggtccgaaat gatggaatca tttattccac cagccttacc tttacctaca caccagaacc
     2161 agggccgcgg ccacattgca gtgcagcagg agcaatcctt cgagccaatt caagccaggt
     2221 gccccctaac gaatcaaaca caaacagcga gggaagttac acaaacgcca gcacaaattc
     2281 aaccagtgtc acatcatcta cagccacagt ggtatcctaa ctaccgtctt tttgctagga
     2341 cttaaactga cttgagtgtg gcaaaaagtt aacaaaaaag gagaaaaaat gaacaatcgt
     2401 ttgtggtttc ttgggaaaac ttttcatacc aggtgatact attcaaaaac cccgttgtct
     2461 ccctgcaagt gctgatttga aatgcagaag ccacagtaaa aaaaaaaaaa aaaaaactcg
     2521 agcatgcatc tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca
     2581 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2641 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2701 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2761 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     2821 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     2881 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgctag cgccctagcg
     2941 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3001 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3061 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3121 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3181 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3241 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3301 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3361 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3421 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3481 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3541 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3601 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3661 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3721 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3781 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3841 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3901 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3961 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4021 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4081 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4141 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4201 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     4261 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4321 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4381 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4441 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4501 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4561 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4621 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4681 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4741 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     4801 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4861 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4921 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4981 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5041 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5101 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5161 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5221 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5281 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5341 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5401 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5461 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5521 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5581 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5641 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5701 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5761 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5821 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5881 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5941 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6001 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6061 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6121 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6181 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6241 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6301 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6361 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6421 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6481 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6541 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6601 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6661 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6721 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6781 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6841 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6901 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6961 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     7021 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta t
//
