LOCUS       sequence-331157-        7070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of CSL F235R.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7070)
  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 7070)
  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 F235R
FEATURES             Location/Qualifiers
     source          1..7070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        130..509
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        510..713
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     664..684
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     758..777
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        758..776
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             799..822
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2518..2536)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2519..2536)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2556..2573)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2562..2786
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2832..3260
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2919..2938)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3129..3150
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3269..3289)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3274..3603
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3454..3589
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3516..3535
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3670..4464
                     /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(3724..3743)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4334..4353
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4638..4759
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4675..4694)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4729..4748
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4808..4824)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4808..4824)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4821..4843)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4832..4848
                     /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(4856..4862,4863..4880,4881..4886))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4901..4922
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5039..5056)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5210..5798)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5290..5309)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5969..6760,6761..6829))
                     /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     6592..6611
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6830..6934)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7009..7028)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
       61 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
      121 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      181 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      241 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      301 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      361 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      421 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      481 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      541 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      601 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      661 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      721 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      781 cccaagcttg gtaccatgga ctacaaagac gatgacgata aaggaattct gcagatgcct
      841 gttgtgacag ggaaatttgg tgagcggcct ccacctaaac gacttactag ggaagctatg
      901 cgaaattatt taaaagagcg aggggatcaa acagtactta ttcttcatgc aaaagttgca
      961 cagaagtcat atggaaatga aaaaaggttt ttttgcccac ctccttgtgt atatcttatg
     1021 ggcagtggat ggaagaaaaa aaaagaacaa atggaacgcg atggttgttc tgaacaagag
     1081 tctcaaccgt gtgcatttat tgggatagga aatagtgacc aagaaatgca gcagctaaac
     1141 ttggaaggaa agaactattg cacagccaaa acattgtata tatctgactc agacaagcga
     1201 aagcacttca tgttgtctgt aaagatgttc tatggcaaca gtgatgacat tggtgtgttc
     1261 ctcagcaagc ggataaaagt catctccaaa ccttccaaaa agaagcagtc attgaaaaat
     1321 gctgacttat gcattgcctc aggaacaaag gtggctctgt ttaatcgact acgatcccag
     1381 acagttagta ccagatactt gcatgtagaa ggaggtaatt ttcatgccag ttcacagcag
     1441 tggggagcct tttttattca tctcttggat gatgatgaat cagaaggaga agaacgcaca
     1501 gtccgagatg gctacatcca ttatggacaa acagtcaaac ttgtgtgctc agttactggc
     1561 atggcactcc caagattgat aattaggaaa gttgataagc agaccgcatt attggatgca
     1621 gatgatcctg tgtcacaact ccataaatgt gcattttacc ttaaggatac agaaagaatg
     1681 tatttgtgcc tttctcaaga aagaataatt caatttcagg ccactccatg tccaaaagaa
     1741 ccaaataaag agatgataaa tgatggcgct tcctggacaa tcattagcac agataaggca
     1801 gagtatacat tttatgaggg aatgggccct gtccttgccc cagtcactcc tgtgcctgtg
     1861 gtagagagcc ttcagttgaa tggcggtggg gacgtagcaa tgcttgaact tacaggacag
     1921 aatttcactc caaatttacg agtgtggttt ggggatgtag aagctgaaac tatgtacagg
     1981 tgtggagaga gtatgctctg tgtcgtccca gacatttctg cattccgaga aggttggaga
     2041 tgggtccggc aaccagtcca ggttccagta actttggtcc gaaatgatgg aatcatttat
     2101 tccaccagcc ttacctttac ctacacacca gaaccagggc cgcggccaca ttgcagtgca
     2161 gcaggagcaa tccttcgagc caattcaagc caggtgcccc ctaacgaatc aaacacaaac
     2221 agcgagggaa gttacacaaa cgccagcaca aattcaacca gtgtcacatc atctacagcc
     2281 acagtggtat cctaactacc gtctttttgc taggacttaa actgacttga gtgtggcaaa
     2341 aagttaacaa aaaaggagaa aaaatgaaca atcgtttgtg gtttcttggg aaaacttttc
     2401 ataccaggtg atactattca aaaaccccgt tgtctccctg caagtgctga tttgaaatgc
     2461 agaagccaca gtaaaaaaaa aaaaaaaaaa ctcgagcatg catctagagg gccctattct
     2521 atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca
     2581 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2641 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2701 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     2761 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     2821 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2881 cagcgtgacc gctacacttg ctagcgccct agcgcccgct cctttcgctt tcttcccttc
     2941 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3001 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3061 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3121 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3181 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3241 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3301 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3361 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3421 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3481 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3541 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3601 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     3661 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3721 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3781 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3841 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3901 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3961 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4021 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4081 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4141 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4201 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4261 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4321 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4381 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4441 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4501 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4561 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4621 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4681 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4741 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4801 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4861 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4921 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4981 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5041 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5101 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5161 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5221 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5281 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5341 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5401 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5461 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5521 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5581 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5641 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5701 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5761 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5821 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5881 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5941 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6001 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6061 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6121 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6181 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6241 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6301 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6361 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6421 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6481 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6541 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6601 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6661 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6721 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6781 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6841 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6901 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6961 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
     7021 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
//
