LOCUS       sequence-331175-        7071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of CSL Q307R.
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  Q307R
FEATURES             Location/Qualifiers
     source          1..7071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        171..550
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        551..754
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     705..725
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     799..818
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        799..817
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             840..863
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2560..2578)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2561..2578)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2598..2615)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2604..2828
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2874..3302
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2961..2980)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3171..3192
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3311..3331)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3316..3645
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3496..3631
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3558..3577
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3712..4506
                     /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(3766..3785)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4376..4395
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4680..4801
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4717..4736)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4771..4790
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4850..4866)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4850..4866)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4863..4885)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4874..4890
                     /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(4898..4904,4905..4922,4923..4928))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4943..4964
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5081..5098)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5252..5840)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5332..5351)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6011..6802,6803..6871))
                     /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     6634..6653
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6872..6976)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7051..7070)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
       61 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
      121 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt
      181 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      241 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      301 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      361 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      421 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      481 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      541 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      601 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      661 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      721 gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc
      781 ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt ggtaccatgg
      841 actacaaaga cgatgacgat aaaggaattc tgcagatgcc tgttgtgaca gggaaatttg
      901 gtgagcggcc tccacctaaa cgacttacta gggaagctat gcgaaattat ttaaaagagc
      961 gaggggatca aacagtactt attcttcatg caaaagttgc acagaagtca tatggaaatg
     1021 aaaaaaggtt tttttgccca cctccttgtg tatatcttat gggcagtgga tggaagaaaa
     1081 aaaaagaaca aatggaacgc gatggttgtt ctgaacaaga gtctcaaccg tgtgcattta
     1141 ttgggatagg aaatagtgac caagaaatgc agcagctaaa cttggaagga aagaactatt
     1201 gcacagccaa aacattgtat atatctgact cagacaagcg aaagcacttc atgttgtctg
     1261 taaagatgtt ctatggcaac agtgatgaca ttggtgtgtt cctcagcaag cggataaaag
     1321 tcatctccaa accttccaaa aagaagcagt cattgaaaaa tgctgactta tgcattgcct
     1381 caggaacaaa ggtggctctg tttaatcgac tacgatccca gacagttagt accagatact
     1441 tgcatgtaga aggaggtaat tttcatgcca gttcacagca gtggggagcc ttttttattc
     1501 atctcttgga tgatgatgaa tcagaaggag aagaattcac agtccgagat ggctacatcc
     1561 attatggaca aacagtcaaa cttgtgtgct cagttactgg catggcactc ccaagattga
     1621 taattaggaa agttgataag cagaccgcat tattggatgc agatgatcct gtgtcacaac
     1681 tccataaatg tgcattttac cttaaggata cagaaagaat gtatttgtgc ctttctcaag
     1741 aaagaataat tcgatttcag gccactccat gtccaaaaga accaaataaa gagatgataa
     1801 atgatggcgc ttcctggaca atcattagca cagataaggc agagtataca ttttatgagg
     1861 gaatgggccc tgtccttgcc ccagtcactc ctgtgcctgt ggtagagagc cttcagttga
     1921 atggcggtgg ggacgtagca atgcttgaac ttacaggaca gaatttcact ccaaatttac
     1981 gagtgtggtt tggggatgta gaagctgaaa ctatgtacag gtgtggagag agtatgctct
     2041 gtgtcgtccc agacatttct gcattccgag aaggttggag atgggtccgg caaccagtcc
     2101 aggttccagt aactttggtc cgaaatgatg gaatcattta ttccaccagc cttaccttta
     2161 cctacacacc agaaccaggg ccgcggccac attgcagtgc agcaggagca atccttcgag
     2221 ccaattcaag ccaggtgccc cctaacgaat caaacacaaa cagcgaggga agttacacaa
     2281 acgccagcac aaattcaacc agtgtcacat catctacagc cacagtggta tcctaactac
     2341 cgtctttttg ctaggactta aactgacttg agtgtggcaa aaagttaaca aaaaaggaga
     2401 aaaaatgaac aatcgtttgt ggtttcttgg gaaaactttt cataccaggt gatactattc
     2461 aaaaaccccg ttgtctccct gcaagtgctg atttgaaatg cagaagccac agtaaaaaaa
     2521 aaaaaaaaaa aactcgagca tgcatctaga gggccctatt ctatagtgtc acctaaatgc
     2581 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2641 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2701 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2761 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     2821 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     2881 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2941 tgctagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3001 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3061 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3121 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3181 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3241 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3301 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3361 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3421 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3481 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3541 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3601 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     3661 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3721 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3781 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3841 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3901 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3961 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4021 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4081 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4141 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4201 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4261 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4321 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4381 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4441 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4501 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4561 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4621 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4681 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4741 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4801 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4861 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4921 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4981 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5041 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5101 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5161 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5221 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5281 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5341 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5401 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5461 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5521 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5581 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5641 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5701 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5761 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5821 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5881 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5941 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6001 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6061 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6121 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6181 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6241 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6301 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6361 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6421 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6481 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6541 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6601 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6661 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6721 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6781 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6841 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6901 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6961 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     7021 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg c
//
