LOCUS       sequence-429467-        7749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  homologous recombination donor for the insertion of the 3xMS2-tag at
            the endogenous locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7749)
  AUTHORS   Klump BM, Perez GI, Patrick EM, Adams-Boone K, Cohen SB, Han L, Yu 
            K, Schmidt JC
  TITLE     TCAB1 prevents nucleolar accumulation of the telomerase RNA to 
            facilitate telomerase assembly.
  JOURNAL   Cell Rep. 2023 Jun 1;42(6):112577. doi: 
            10.1016/j.celrep.2023.112577.
  PUBMED    37267110
REFERENCE   2  (bases 1 to 7749)
  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: "Cell Rep.";
            date: "2023-06-1"; volume: "42(6):112577. doi"; pages: " 
            10.1016/j.celrep.2023.112577"
COMMENT     Sequence Label: 3xMS-TR-100-PURO HRD
FEATURES             Location/Qualifiers
     source          1..7749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..135
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(38..57)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     92..111
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  164..329
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      513..967
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(600..619)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     809..830
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        993..1097
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1098..1166,1167..1958)
                     /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     complement(1316..1335)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2129..2717
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2618..2637
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2871..2888
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  3022..3246
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(3313..3846)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4035..4040,4041..4057,4058..4063))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    4571..4619
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4644..4663
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(4755..4864)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        4888..4979
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4904..4921
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     misc_RNA        5667..5685
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        5718..5736
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     promoter        6306..6622
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6473..6608
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6629..7228
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6629..6648)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7125..7145
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7229..7277
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
       61 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
      121 tatcatgtct ggatctgatc actgcttgag cctaggagat ccgaaccaga taagtgaaat
      181 ctagttccaa actattttgt catttttaat tttcgtatta gcttacgacg ctacacccag
      241 ttcccatcta ttttgtcact cttccctaaa taatccttaa aaactccatt tccacccctc
      301 ccagttccca actattttgt ccgcccacag cggggcattt ttcttcctgt tatgttttta
      361 atcaaacatc ctgccaactc catgtgacaa accgtcatct tcggctactt tttctctgtc
      421 acagaatgaa aatttttctg tcatctcttc gttattaatg tttgtaattg actgaatatc
      481 aacgcttatt tgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
      541 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
      601 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
      661 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
      721 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
      781 tttgacgttg gagtccacgt tcttaatagt ggactcttgt tccaaactgg aacaacactc
      841 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
      901 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
      961 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1021 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1081 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     1141 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     1201 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     1261 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     1321 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     1381 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     1441 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     1501 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     1561 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     1621 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     1681 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     1741 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     1801 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     1861 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     1921 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     1981 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     2041 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2101 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2161 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2221 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     2281 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2341 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2401 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2461 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2521 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2581 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2641 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2701 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     2761 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     2821 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     2881 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     2941 tttcacaccg catagaccag ccgcgtaacc tggcaaaatc ggttacggtt gagtaataaa
     3001 tggatgccct gcgtaagcgg gtgtgggcgg acaataaagt cttaaactga acaaaataga
     3061 tctaaactat gacaataaag tcttaaacta gacagaatag ttgtaaactg aaatcagtcc
     3121 agttatgctg tgaaaaagca tactggactt ttgttatggc taaagcaaac tcttcatttt
     3181 ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg ccaagggcat ggtaaagact
     3241 atattcgcgg cgttgtgaca atttaccgaa caactccgcg gccgggaagc cgatctcggc
     3301 ttgaacgaat tgttaggtgg cggtacttgg gtcgatatca aagtgcatca cttcttcccg
     3361 tatgcccaac tttgtataga gagccactgc gggatcgtca ccgtaatctg cttgcacgta
     3421 gatcacataa gcaccaagcg cgttggcctc atgcttgagg agattgatga gcgcggtggc
     3481 aatgccctgc ctccggtgct cgccggagac tgcgagatca tagatataga tctcactacg
     3541 cggctgctca aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg cttcttggtc
     3601 gaaggcagca agcgcgatga atgtcttact acggagcaag ttcccgaggt aatcggagtc
     3661 cggctgatgt tgggagtagg tggctacgtc tccgaactca cgaccgaaaa gatcaagagc
     3721 agcccgcatg gatttgactt ggtcagggcc gagcctacat gtgcgaatga tgcccatact
     3781 tgagccacct aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctgcg
     3841 taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct
     3901 tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg aaaaccgcca
     3961 ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac
     4021 gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt cgtgccttca
     4081 tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc gaggcatttc
     4141 tgtcctggct ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag gcattggcgg
     4201 ccttgctgtt cttctacggc aaggtgctgt gcacggatct gcccttgctt caggagatcg
     4261 gtagacctcg gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa gtggttcgca
     4321 tcctcggttt tctggaaggc gagcatcgtt tgttcgccca ggactctagc tatagttcta
     4381 gtggttggct acgtacccgt agtggctatg gcagggcttg ccgccccgac gttggctgcg
     4441 agccctgggc cttcacccga acttgggggt tggggtgggg aaaaggaaga aacgcgggcg
     4501 tattggtccc aatggggtct cggtggggta tcgacagagt gccagccctg ggaccgaacc
     4561 ccgcgtttat gaacaaacga cccaacaccc gtgcgtttta ttctgtcttt ttattgccgt
     4621 catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc tcccccatct
     4681 cccggtaccg catgctatgc atcagctgct agcaccatgg ctcgagatcc cgggtgatca
     4741 agtcttcgtc gagtgattgt aaataaaatg taatttacag tatagtattt taattaatat
     4801 acaaatgatt tgataataat tcttatttaa ctataatata ttgtgttggg ttgaattaaa
     4861 ggtccgtata ctccggaata ttaatagatc atggagataa ttaaaatgat aaccatctcg
     4921 caaataaata agtattttac tgttttcgta acagttttgt aataaaaaaa cctataaata
     4981 ttccggatta ttcataccgt cccaccatcg ggcgcggatc ccggtccgaa gcgcgcggaa
     5041 ttcaaaggcc tacgtcgacg agctcactag tcgcggccgc tttcgaatct agagcctgca
     5101 gtctcgacaa gcttgtcgag aagtactaga ggatcataat cagccatacc acatttgtag
     5161 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     5221 atgcaattgt tgttgtttct gctgtagtca gtgctgcctg ggaatctatt ttcacaaagt
     5281 tctccaaaaa atgtgatgat caaaactagg aattagtgtt ctgtgtctta ggccctaaaa
     5341 tcttcctgtg aattccattt ttaaggtagt cgaggtgaac cgcgtctggt ctgcagagga
     5401 tagaaaaaag gccctctgat acctcaagtt agtttcacct ttaaagaagg tcggaagtaa
     5461 agacgcaaag cctttcccgg acgtgcggaa gggcaacgtc cttcctcatg gccggaaatg
     5521 gaactttaat ttcccgttcc ccccaaccag cccgcccgag agagtgactc tcacgagagc
     5581 cgcgagagtc agcttggcca atccgtgcgg tcggcggccg ctccctttat aagccgactc
     5641 gcccggcagc gcacccggac gagctgacat gaggatcacc catgtctgca gcgacgaagg
     5701 atcacctcgt cgataaaaca tgaggatcac ccatgtctgc aggcagctcg tccggggttg
     5761 cggagggtgg gcctgggagg ggtggtggcc attttttgtc taaccctaac tgagaagggc
     5821 gtaggcgccg tgcttttgct ccccgcgcgc tgtttttctc gctgactttc agcgggcgga
     5881 aaagcctcgg cctgccgcct tccaccgttc attctagagc aaacaaaaaa tgtcagctgc
     5941 tggcccgttc gcccctcccg gggacctgcg gcgggtcgcc tgcccagccc ccgaaccccg
     6001 cctggaggcc gcggtcggcc cggggcttct ccggaggcac ccactgccac cgcgaagagt
     6061 tgggctctgt cagccgcggg tctctcgggg gcgagggcga ggttcaggcc tttcaggccg
     6121 caggaagagg aacggagcga gtccccgcgc gcggcgcgat tccctgagct gtgggacgtg
     6181 cacccaggac tcggctcaca catgcagttc gctttcctgt tggtgggggg aacgccgatc
     6241 gtgcgcatcc gtcacccctc gccggcagtg ggggcttgtg aacccccaaa cctgactgac
     6301 taaacggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6361 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     6421 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     6481 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     6541 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     6601 tggaggccta ggcttttgca aaaagctcat gaccgagtac aagcccacgg tgcgcctcgc
     6661 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     6721 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     6781 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     6841 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     6901 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     6961 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     7021 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     7081 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     7141 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     7201 gtgcatgacc cgcaagcccg gtgcctgaaa taaaatatct ttattttcat tacatctgtg
     7261 tgttggtttt ttgtgtgcag tgtgctgcaa attggcagga gacgtgaagg cacctccaaa
     7321 gtcggccaaa atgaatgggc agtgagccgg ggttgcctgg agccgttcct gcgtgggttc
     7381 tcccgtcttc cgctttttgt tgccttttat ggttgtatta caacttagtt cctgctctgc
     7441 agattttgtt gaggtttttg cttctcccaa ggtagatctc gaccagtccc ctcaacgggg
     7501 tgtgggagaa cagtcatttt ttttgagaga tcatttaaca tttaatgaat atttaattag
     7561 aagatctaaa tgaacattgg aaattgtgtt cctttaatgg tcatcggttt atgccagagg
     7621 ttagaagttt cttttttgaa aaattagacc ttggcgatga ccttgagcag taggatataa
     7681 cccccacaag cttagcgttc caataacgga acactaggca taatgaaaga cggaaaagaa
     7741 atttattcc
//
