LOCUS       sequence-331145-        7071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of CSL WT.
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 WT
FEATURES             Location/Qualifiers
     source          1..7071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        139..518
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        519..722
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     673..693
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     767..786
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        767..785
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             808..831
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2528..2546)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2529..2546)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2566..2583)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2572..2796
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2842..3270
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2929..2948)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3139..3160
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3279..3299)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3284..3613
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3464..3599
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3526..3545
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3680..4474
                     /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(3734..3753)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4344..4363
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4648..4769
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4685..4704)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4739..4758
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4818..4834)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4818..4834)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4831..4853)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4842..4858
                     /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(4866..4872,4873..4890,4891..4896))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4911..4932
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5049..5066)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5220..5808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5300..5319)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5979..6770,6771..6839))
                     /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     6602..6621
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6840..6944)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7019..7038)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
       61 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
      121 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
      181 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      241 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      301 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      361 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      421 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      481 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      541 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      601 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      661 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      721 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
      781 atagggagac ccaagcttgg taccatggac tacaaagacg atgacgataa aggaattctg
      841 cagatgcctg ttgtgacagg gaaatttggt gagcggcctc cacctaaacg acttactagg
      901 gaagctatgc gaaattattt aaaagagcga ggggatcaaa cagtacttat tcttcatgca
      961 aaagttgcac agaagtcata tggaaatgaa aaaaggtttt tttgcccacc tccttgtgta
     1021 tatcttatgg gcagtggatg gaagaaaaaa aaagaacaaa tggaacgcga tggttgttct
     1081 gaacaagagt ctcaaccgtg tgcatttatt gggataggaa atagtgacca agaaatgcag
     1141 cagctaaact tggaaggaaa gaactattgc acagccaaaa cattgtatat atctgactca
     1201 gacaagcgaa agcacttcat gttgtctgta aagatgttct atggcaacag tgatgacatt
     1261 ggtgtgttcc tcagcaagcg gataaaagtc atctccaaac cttccaaaaa gaagcagtca
     1321 ttgaaaaatg ctgacttatg cattgcctca ggaacaaagg tggctctgtt taatcgacta
     1381 cgatcccaga cagttagtac cagatacttg catgtagaag gaggtaattt tcatgccagt
     1441 tcacagcagt ggggagcctt ttttattcat ctcttggatg atgatgaatc agaaggagaa
     1501 gaattcacag tccgagatgg ctacatccat tatggacaaa cagtcaaact tgtgtgctca
     1561 gttactggca tggcactccc aagattgata attaggaaag ttgataagca gaccgcatta
     1621 ttggatgcag atgatcctgt gtcacaactc cataaatgtg cattttacct taaggataca
     1681 gaaagaatgt atttgtgcct ttctcaagaa agaataattc aatttcaggc cactccatgt
     1741 ccaaaagaac caaataaaga gatgataaat gatggcgctt cctggacaat cattagcaca
     1801 gataaggcag agtatacatt ttatgaggga atgggccctg tccttgcccc agtcactcct
     1861 gtgcctgtgg tagagagcct tcagttgaat ggcggtgggg acgtagcaat gcttgaactt
     1921 acaggacaga atttcactcc aaatttacga gtgtggtttg gggatgtaga agctgaaact
     1981 atgtacaggt gtggagagag tatgctctgt gtcgtcccag acatttctgc attccgagaa
     2041 ggttggagat gggtccggca accagtccag gttccagtaa ctttggtccg aaatgatgga
     2101 atcatttatt ccaccagcct tacctttacc tacacaccag aaccagggcc gcggccacat
     2161 tgcagtgcag caggagcaat ccttcgagcc aattcaagcc aggtgccccc taacgaatca
     2221 aacacaaaca gcgagggaag ttacacaaac gccagcacaa attcaaccag tgtcacatca
     2281 tctacagcca cagtggtatc ctaactaccg tctttttgct aggacttaaa ctgacttgag
     2341 tgtggcaaaa agttaacaaa aaaggagaaa aaatgaacaa tcgtttgtgg tttcttggga
     2401 aaacttttca taccaggtga tactattcaa aaaccccgtt gtctccctgc aagtgctgat
     2461 ttgaaatgca gaagccacag taaaaaaaaa aaaaaaaaaa ctcgagcatg catctagagg
     2521 gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt
     2581 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     2641 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     2701 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     2761 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     2821 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2881 tggttacgcg cagcgtgacc gctacacttg ctagcgccct agcgcccgct cctttcgctt
     2941 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3001 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3061 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3121 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3181 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3241 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     3301 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3361 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3421 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3481 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3541 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3601 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     3661 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3721 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3781 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3841 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3901 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3961 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4021 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4081 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4141 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4201 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4261 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4321 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4381 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4441 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4501 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4561 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4621 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     4681 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4741 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     4801 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4861 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4921 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4981 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5041 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5101 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5161 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5221 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5281 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5341 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5401 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5461 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5521 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5581 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5641 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5701 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5761 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5821 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5881 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5941 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6001 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6061 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6121 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6181 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6241 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6301 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6361 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6421 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6481 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6541 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6601 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6661 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6721 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6781 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6841 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6901 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6961 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     7021 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt g
//
