LOCUS       sequence-331174-        7071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of CSL A258R.
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  A258R
FEATURES             Location/Qualifiers
     source          1..7071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        159..538
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        539..742
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     693..713
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     787..806
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        787..805
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             828..851
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2548..2566)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2549..2566)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2586..2603)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2592..2816
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2862..3290
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2949..2968)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3159..3180
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3299..3319)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3304..3633
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3484..3619
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3546..3565
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3700..4494
                     /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(3754..3773)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4364..4383
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4668..4789
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4705..4724)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4759..4778
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4838..4854)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4838..4854)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4851..4873)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4862..4878
                     /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(4886..4892,4893..4910,4911..4916))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4931..4952
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5069..5086)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5240..5828)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5320..5339)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5999..6790,6791..6859))
                     /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     6622..6641
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6860..6964)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7039..7058)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
       61 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
      121 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
      181 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      241 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      301 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      361 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      421 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      481 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      541 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      601 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      661 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      721 ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt actggcttat
      781 cgaaattaat acgactcact atagggagac ccaagcttgg taccatggac tacaaagacg
      841 atgacgataa aggaattctg cagatgcctg ttgtgacagg gaaatttggt gagcggcctc
      901 cacctaaacg acttactagg gaagctatgc gaaattattt aaaagagcga ggggatcaaa
      961 cagtacttat tcttcatgca aaagttgcac agaagtcata tggaaatgaa aaaaggtttt
     1021 tttgcccacc tccttgtgta tatcttatgg gcagtggatg gaagaaaaaa aaagaacaaa
     1081 tggaacgcga tggttgttct gaacaagagt ctcaaccgtg tgcatttatt gggataggaa
     1141 atagtgacca agaaatgcag cagctaaact tggaaggaaa gaactattgc acagccaaaa
     1201 cattgtatat atctgactca gacaagcgaa agcacttcat gttgtctgta aagatgttct
     1261 atggcaacag tgatgacatt ggtgtgttcc tcagcaagcg gataaaagtc atctccaaac
     1321 cttccaaaaa gaagcagtca ttgaaaaatg ctgacttatg cattgcctca ggaacaaagg
     1381 tggctctgtt taatcgacta cgatcccaga cagttagtac cagatacttg catgtagaag
     1441 gaggtaattt tcatgccagt tcacagcagt ggggagcctt ttttattcat ctcttggatg
     1501 atgatgaatc agaaggagaa gaattcacag tccgagatgg ctacatccat tatggacaaa
     1561 cagtcaaact tgtgtgctca gttactggca tgcgactccc aagattgata attaggaaag
     1621 ttgataagca gaccgcatta ttggatgcag atgatcctgt gtcacaactc cataaatgtg
     1681 cattttacct taaggataca gaaagaatgt atttgtgcct ttctcaagaa agaataattc
     1741 aatttcaggc cactccatgt ccaaaagaac caaataaaga gatgataaat gatggcgctt
     1801 cctggacaat cattagcaca gataaggcag agtatacatt ttatgaggga atgggccctg
     1861 tccttgcccc agtcactcct gtgcctgtgg tagagagcct tcagttgaat ggcggtgggg
     1921 acgtagcaat gcttgaactt acaggacaga atttcactcc aaatttacga gtgtggtttg
     1981 gggatgtaga agctgaaact atgtacaggt gtggagagag tatgctctgt gtcgtcccag
     2041 acatttctgc attccgagaa ggttggagat gggtccggca accagtccag gttccagtaa
     2101 ctttggtccg aaatgatgga atcatttatt ccaccagcct tacctttacc tacacaccag
     2161 aaccagggcc gcggccacat tgcagtgcag caggagcaat ccttcgagcc aattcaagcc
     2221 aggtgccccc taacgaatca aacacaaaca gcgagggaag ttacacaaac gccagcacaa
     2281 attcaaccag tgtcacatca tctacagcca cagtggtatc ctaactaccg tctttttgct
     2341 aggacttaaa ctgacttgag tgtggcaaaa agttaacaaa aaaggagaaa aaatgaacaa
     2401 tcgtttgtgg tttcttggga aaacttttca taccaggtga tactattcaa aaaccccgtt
     2461 gtctccctgc aagtgctgat ttgaaatgca gaagccacag taaaaaaaaa aaaaaaaaaa
     2521 ctcgagcatg catctagagg gccctattct atagtgtcac ctaaatgcta gagctcgctg
     2581 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2641 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2701 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2761 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     2821 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     2881 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ctagcgccct
     2941 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3001 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3061 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3121 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3181 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3241 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     3301 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3361 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3421 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3481 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3541 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     3601 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     3661 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     3721 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3781 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3841 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     3901 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3961 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4021 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4081 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4141 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4201 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4261 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4321 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4381 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4441 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     4501 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     4561 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     4621 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     4681 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4741 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     4801 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4861 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     4921 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     4981 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5041 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5101 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5161 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5221 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5281 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5341 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5401 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5461 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5521 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5581 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5641 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5701 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5761 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5821 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5881 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5941 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6001 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6061 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6121 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6181 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6241 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6301 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6361 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6421 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6481 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6541 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6601 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6661 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6721 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6781 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6841 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6961 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     7021 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc a
//
