LOCUS       sequence-351668-        5909 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a unique yeast telomere end at TEL11R by removing the 
            subtelomere.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5909)
  AUTHORS   Sholes SL, Karimian K, Gershman A, Kelly TJ, Timp W, Greider CW
  TITLE     Chromosome-specific telomere lengths and the minimal functional 
            telomere revealed by nanopore sequencing.
  JOURNAL   Genome Res. 2022 Apr;32(4):616-628. doi: 10.1101/gr.275868.121. Epub
            2021 Oct 26.
  PUBMED    34702734
REFERENCE   2  (bases 1 to 5909)
  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"; doi: 
            "10.1101/gr.275868.121"; journalName: "Genome Res"; date: "2022-04";
            volume: "32"; issue: "4"; pages: "616-628"
COMMENT     Sequence Label: pCS105
FEATURES             Location/Qualifiers
     source          1..5909
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     16..38
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     30..47
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     31..47
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     54..73
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        54..72
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        183..590
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             591..1685
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     terminator      1686..1933
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1702..1720)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2194..2210)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2194..2210)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2207..2229)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2218..2234
                     /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(2242..2248,2249..2266,2267..2272))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2287..2308
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2425..2442)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2596..3184)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2676..2695)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3355..4146,4147..4215))
                     /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     3978..3997
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(4233..5027)
                     /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(4344..4363)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4954..4973
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      complement(5371..5799)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5481..5502)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5693..5712
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg
       61 actcactata gggcgaattg ggccctctag atgcatgctc gagcggccgc cagtgtgatg
      121 gatatctgca gaattcgccc ttgctggtac attttatggt atataaacaa gctacgttat
      181 taaactgtgg gaatactcag gtatcgtaag atgcaagagt tcgaatctct tagcaaccat
      241 tatttttttc ctcaacataa cgagaacaca caggggcgct atcgcacaga atcaaattcg
      301 atgactggaa attttttgtt aatttcagag gtcgcctgac gcatatacct ttttcaactg
      361 aaaaattggg agaaaaagga aaggtgagag cgccggaacc ggcttttcat atagaataga
      421 gaagcgttca tgactaaatg cttgcatcac aatacttgaa gttgacaata ttatttaagg
      481 acctattgtt ttttccaata ggtggttagc aatcgtctta ctttctaact tttcttacct
      541 tttacatttc agcaatatat atatatatat ttcaaggata taccattcta atgtctgccc
      601 ctaagaagat cgtcgttttg ccaggtgacc acgttggtca agaaatcaca gccgaagcca
      661 ttaaggttct taaagctatt tctgatgttc gttccaatgt caagttcgat ttcgaaaatc
      721 atttaattgg tggtgctgct atcgatgcta caggtgttcc acttccagat gaggcgctgg
      781 aagcctccaa gaaggctgat gccgttttgt taggtgctgt gggtggtcct aaatggggta
      841 ccggtagtgt tagacctgaa caaggtttac taaaaatccg taaagaactt caattgtacg
      901 ccaacttaag accatgtaac tttgcatccg actctctttt agacttatct ccaatcaagc
      961 cacaatttgc taaaggtact gacttcgttg ttgtcagaga attagtggga ggtatttact
     1021 ttggtaagag aaaggaagac gatggtgatg gtgtcgcttg ggatagtgaa caatacaccg
     1081 ttccagaagt gcaaagaatc acaagaatgg ccgctttcat ggccctacaa catgagccac
     1141 cattgcctat ttggtccttg gataaagcta atgttttggc ctcttcaaga ttatggagaa
     1201 aaactgtgga ggaaaccatc aagaacgaat tccctacatt gaaggttcaa catcaattga
     1261 ttgattctgc cgccatgatc ctagttaaga acccaaccca cctaaatggt attataatca
     1321 ccagcaacat gtttggtgat atcatctccg atgaagcctc cgttatccca ggttccttgg
     1381 gtttgttgcc atctgcgtcc ttggcctctt tgccagacaa gaacaccgca tttggtttgt
     1441 acgaaccatg ccacggttct gctccagatt tgccaaagaa taaggtcaac cctatcgcca
     1501 ctatcttgtc agctgcaatg atgttgaaat tgtcattgaa cttgcctgaa gaaggtaagg
     1561 ccattgaaga tgcagttaaa aaggttttgg atgcaggtat cagaactggt gatttaggtg
     1621 gttccaacag taccaccgaa gtcggtgatg ctgtcgccga agaagttaag aaaatccttg
     1681 cttaatcatg taattagtta tgtcacgctt acattcacgc cctcccccca catccgctct
     1741 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt tttttatagt
     1801 tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacagacg
     1861 cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg gacgctcgaa
     1921 ggctttaatt tgccggagta ctgtcctccg agcggtgaac tgtcctccga ctcgtgcgga
     1981 gtactgtcct ccgatcggtg aactgtcctc cgcgaattcc ggagtactaa tctccgggtg
     2041 gtgtgggtgt ggtgtgtggg tgtggtgtgt ggtgtgtagg gataacaggg taataagggc
     2101 gaattctgca gatatccatc aagggcgaat tccagcacac tggcggccgt tactagtgga
     2161 tccgagctcg gtaccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     2221 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     2281 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     2341 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2401 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2461 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2521 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2581 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2641 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2701 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2761 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2821 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2881 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2941 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3001 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3061 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3121 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3181 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3241 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3301 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3361 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3421 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3481 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3541 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3601 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3661 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3721 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3781 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3841 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3901 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3961 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4021 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4081 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4141 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4201 atgttgaata ctcatactct tcctttttca attcagaaga actcgtcaag aaggcgatag
     4261 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
     4321 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg
     4381 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
     4441 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgcgc
     4501 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
     4561 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
     4621 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
     4681 atgatggata ctttctcggc aggagcaagg tgggatgaca ggagatcctg ccccggcact
     4741 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     4801 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg
     4861 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     4921 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     4981 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct
     5041 catcctgtct cttgatcaga tcttgatccc ctgcgccatc agatccttgg cggcaagaaa
     5101 gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat
     5161 tccggttcgc ttgctgtcca taaaaccgcc cagtctagct atcgccatgt aagcccactg
     5221 caagctacct gctttctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg
     5281 acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacgtgtt ccgcttcctt
     5341 tagcagccct tgcgccctga attttgttaa aattcgcgtt aaatttttgt taaatcagct
     5401 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     5461 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     5521 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     5581 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     5641 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     5701 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     5761 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg
     5821 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     5881 gggatgtgct gcaaggcgat taagttggg
//
