LOCUS       sequence-137330-        6844 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag-tagged mRAP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6844)
  AUTHORS   Sfeir A, Kabir S, van Overbeek M, Celli GB, de Lange T
  TITLE     Loss of Rap1 induces telomere recombination in the absence of NHEJ 
            or a DNA damage signal.
  JOURNAL   Science. 2010 Mar 26;327(5973):1657-61. doi: 
            10.1126/science.1185100.
  PUBMED    20339076
REFERENCE   2  (bases 1 to 6844)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2010-03-26- 26"; volume: "327"; 
            issue: "5973"; pages: "1657-61"
COMMENT     Sequence Label: pLPC Flag-mRAP1
FEATURES             Location/Qualifiers
     source          1..6844
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             120..589
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(1962..1985)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      1985..1994
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2025..2044)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2026..2044)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2089..2292)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2118..2138)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2293..2672
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2844..2863
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(2952..3551)
                     /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(3035..3055)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3532..3551
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(3569..3985)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3585..3601)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3617..3639)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    4186..4385
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             4450..4919
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        4950..5054
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5055..5123,5124..5915)
                     /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(5273..5292)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6075..6663
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6564..6583
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6817..6834
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaactatg
       61 gccatttaat gtaaatactt aagaaaaaaa accaaattaa ttttgataca tgctgcatgt
      121 gaaagacccc cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga
      181 gaccacaagt cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgact
      241 cagtcaatcg gaggactggc gccccgagtg aggggttgtg ggctctttta ttgagctcgg
      301 ggagcagaag cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca
      361 cagggtcatt tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagaaactgc
      421 tgagggctgg accgcatctg gggaccatct gttcttggcc ctgagccggg gcaggaactg
      481 cttaccacag atatcctgtt tggcccatat tcagctgttc catctgttct tggccctgag
      541 ccggggcagg aactgcttac cacagatatc ctgtttggcc catattcagc tgttccatct
      601 gttcctgacc ttgatctgaa cttctctatt ctcagttatg tatttttcca tgccttgcaa
      661 aatggcgtta cttaagctag cttgccaaac ctacaggtgg ggtctttcat tccccccttt
      721 ttctggagac taaataaaat cttttatttt atcggccctc tagatgcatg ctcgagttat
      781 ttctttcgga attcaatcct ccgagcaacg ttctgagctc caaacttttt gaccagtgca
      841 tttttcgtgt cgtcatcgtc cttctgcaaa tctaagtcat cttgtctgca ccaaatggga
      901 taaccgtcgg gtctccgccc cgactctaag aaggaggacg tggcctccag ctcaccactg
      961 ttcttcagca aggcctgtgt aactgttgat agatccaagt tgaacttctc cattagctgc
     1021 cggatcacct taatggcagt tcccacttct tgcgtagaaa cttttggttc ttcttcctcc
     1081 tcgtctggct gtgtttctga gtcttcctcg ggtgtgggtg gatcaccatc acacatcgtt
     1141 atatgtattt caaagtcagg gctctcatcc accacggctt ctccgaactc cggagcggct
     1201 tcctcaaaca gcttgttgtc tgcctctcca ttctccttga tctctccttt cacacactcc
     1261 tcctcaggca agtctggcgc tctcttgttc tgtggctctc cgctatccgc ggcctcgggg
     1321 tcctgctccg ccttccgttt gagcttctgg gaggacgggc tgaccggggc gttcccgagc
     1381 aggtacttgt gctcctgacc ccgtaggtgc ttgaggtagc ggtccttgag cgactgccag
     1441 gagtgctgcg tgagcgagct cttctccatc gctttccaca aggcattgcc tgtgaccgag
     1501 ctgggcgaac gggcgttctc cttcacgtag gtcaggatgg ccacatcttc cgcgtcggtg
     1561 taggcgatgc gcccggtcag gggctgcggc tccggttccg tggagccctc ggtggaagcc
     1621 ccgggcttcg gatcggacgc ctgctccgtc aggcccagcc gataggcctc caggtccagc
     1681 ttctcgttgc gatccacgca gtctaggatg tactgcgtgg agatgaagtc gcccgaagcc
     1741 tcggccagcg cctccccggg ctgggcgaga agcacggctc cgggttcctg cacccgacac
     1801 acggtgccgc cgccgtgcag gatgagcgtc gacagccggc gcttggccga gctgggccgc
     1861 acgtaaaacg acatggcgct gccgtcttct ctcacgaaca gagtggagga gtgagtgggc
     1921 ccattggggt ctttacccag atccatcgcc tccgcggatc ctttatcgtc atcgtctttg
     1981 tagtccatgg tggctgatcc gagctcggta ccaagcttgg gtctccctat agtgagtcgt
     2041 attaatttcg ataagccagt aagcagtggg ttctctagtt agccagagag ctctgcttat
     2101 atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg
     2161 acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg
     2221 gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca
     2281 aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
     2341 gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
     2401 aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
     2461 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
     2521 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
     2581 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
     2641 ttactattaa taactagtca ataatcaatg tcaacgcgta tatctggccc gtacatcgcg
     2701 aagcagcgca aaacgcctaa ccctaagcag attcttcatg caattgtcgg tcaagccttg
     2761 ccttgttgta gcttaaattt tgctcgcgca ctactcagcg acctccaaca cacaagcagg
     2821 gagcagatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatag
     2881 gggatcggga gatccgatgc atggggtcgt gcgctccttt cggtcgggcg ctgcgggtcg
     2941 tggggcgggc gtcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc
     3001 acctcgacgt cggcggtgac ggtgaagccg agccgctcgt agaaggggag gttgcggggc
     3061 gcggaggtct ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac tccggggagc
     3121 acgacggcgc tgcccagacc cttgccctgg tggtcgggcg agacgccgac ggtggccagg
     3181 aaccacgcgg gctccttggg ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg
     3241 gccagccggg aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa caccgccccc
     3301 gcttcgacgc tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc cgcgacccac
     3361 accttgccga tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc
     3421 tcgatgtggc ggtccggatc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg
     3481 gcgagggtgc gtacggccct ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg
     3541 tactcggtca tggtaagctg atccggccgg cgcctagaga aggagtgagg gctggataaa
     3601 gggaggatcg aggcggggtc gaacgaggag gttcaagggg gagagacggg gcggatggag
     3661 gaagaggagg cggaggctta gggtgtacaa agggcttgac ccagggaggg gggtcaaaag
     3721 ccaaggcttc ccaggtcacg atgtagggga cctggtctgg gtgtccatgc gggccaggtg
     3781 aaaagacctt gatcttaacc tgggtgatga ggtctcggtt aaaggtgccg tctcgcggcc
     3841 atccgacgtt aaaggttggc cattctgcag agcagaaggt aacccaacgt ctcttcttga
     3901 catctaccga ctggttgtga gcgatccgct cgacatcttt ccagtgacct aaggtcaaac
     3961 ttaagggagt ggtaacagtc tggccctaat tttcagacaa atacagaaac acagtcagac
     4021 agagacaaca cagaacgatg ctgcagcgct gcagcagaca agacgcgcgg cttcggttcc
     4081 aaaccgaaag caaaaattca gacggaggcg ggaactgttt taggttctcg tctcctacca
     4141 gaaccacata tcctgacggg gtcggattcc acatcgactc ccttcctcag gtcgggccac
     4201 aaaaacggcc cccaaagtcc ctgggacgtc tcccagggtt gcggccgggt gttcagaact
     4261 cgtcagttcc accacgggtc cgccagatac agagctagtt agctaactag taccgacgca
     4321 ggcgcataaa atcagtcata gacactagac aatcggacag acacagataa gttgctggcc
     4381 agcttgcctc ccggtggtgg gtcggtggtc cctgggcagg ggtctcccga tcccggacga
     4441 gcccccaaat gaaagacccc cgctgacggg tagtcaatca ctcagaggag accctcccaa
     4501 ggaacagcga gaccacaagt cggatgcaac tgcaagaggg tttattggat acacgggtac
     4561 ccgggcgact cagtcaatcg gaggactggc gccccgagtg aggggttgtg ggctctttta
     4621 ttgagctcgg ggagcagaag cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc
     4681 aaataaggca cagggtcatt tcaggtcctt ggggcaccct ggaaacatct gatggttctc
     4741 tagaaactgc tgagggctgg accgcatctg gggaccatct gttcttggcc ctgagccggg
     4801 gcaggaactg cttaccacag atatcctgtt tggcccatat tcagctgttc catctgttct
     4861 tggccctgag ccggggcagg aactgcttac cacagatatc ctgtttggcc catattcagg
     4921 ctgcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4981 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5041 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5101 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5161 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5221 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5281 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5341 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5401 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5461 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5521 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5581 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5641 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5701 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5761 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5821 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5881 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5941 tactttagat tgatttgcgg ccgcaaaagg atctaggtga agatcctttt tgataatctc
     6001 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6061 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6121 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6181 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6241 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6301 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6361 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6421 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6481 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6541 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6601 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6661 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6721 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6781 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6841 gaag
//
