LOCUS       sequence-362653-        7949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of BirA Rap1 I312R.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7949)
  AUTHORS   Barry RM, Sacco O, Mameri A, Stojaspal M, Kartsonis W, Shah P, De 
            Ioannes P, Hofr C, Cote J, Sfeir A
  TITLE     Rap1 regulates TIP60 function during fate transition between 
            two-cell-like and pluripotent states.
  JOURNAL   Genes Dev. 2022 Feb 24. pii: gad.349039.121. doi: 
            10.1101/gad.349039.121.
  PUBMED    35210222
REFERENCE   2  (bases 1 to 7949)
  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: "Genes Dev. 
            2022 Feb 24. pii: gad.349039.121. doi: 10.1101/gad.349039.121."
COMMENT     Sequence Label: pLPC-Flag-BirA-Linker-Myc-mRap1-I312R
FEATURES             Location/Qualifiers
     source          1..7949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(270..299)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(join(402..1010,1011..1013,1014..1361))
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             complement(1368..1391)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      1391..1400
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1431..1450)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1432..1450)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1495..1698)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1524..1544)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1699..2078
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2250..2269
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(2358..2957)
                     /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(2441..2461)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2938..2957
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(2975..3391)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2991..3007)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3023..3045)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3592..3791
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             3856..4325
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        4356..4460
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4461..4529,4530..5321)
                     /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(4679..4698)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5481..6069
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5970..5989
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6223..6240
                     /label=L4440
                     /note="L4440 vector, forward primer"
     LTR             6370..6839
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 aggatgtact gcgtggagat gaagtcgccc gaagcctcgg ccagcgcctc cccgggctgg
       61 gcgagaagca cggctccggg ttcctgcacc cgacacacgg tgccgccgcc gtgcaggatg
      121 agcgtcgaca gccggcgctt ggccgagctg ggccgcacgt aaaacgacat ggcgctgccg
      181 tcttctctca cgaacagagt ggaggagtga gtgggcccat tggggtcttt acccagatcc
      241 atcgcctccg cggatcctct ctttcttaac aaatcttctt cagaaatcaa cttttgttcc
      301 tcgagggagc cccctccgcc agagcctcca ccccctgaac ctccgccacc tgacccgccc
      361 cctccagatc cgccacctcc cgacccaccg cctccgaatt ctttctcggc agatctcagg
      421 ctgatctcgc cgcccatcca gggcttgatg atgccgtcct gttccagcag cagagcgccc
      481 tgcttgtcga tgccccggct gatgccgaag atctctttgt cgccgatgat cagtttcacg
      541 ggtctgttga tgaagttgtc cagcttctcc catctggaca ggtagggggc caggccttcc
      601 tgctcgaaca gttccagggc ggctctcagc tctctgatca gcatggcggc cagggtgttt
      661 ctgtccaggt tgatgccggc ttcctgcagg gtgatccagc cctgattcac gacggactct
      721 tccacccgcc gcatggccat attgatgcca gcgccaatca cgatctgggc agcgtcgcct
      781 gttttgcctg tcagttccac caggatgccg gccagcttcc ggtcctgcag atacaggtcg
      841 ttgggccact tcacgcgcac tttgtcggcg cccagctttc tcagcacctc ggccatcacg
      901 atgccgatca cgaggctcag tccaatagcg gcggcaggtc cctgttccag ccgccagaac
      961 atgctcaggt acagattggc gccgaaggga gagaaccact ttctgcccct gccgcctctg
     1021 ccagcctgct gatactcggc gatacaggcg tcgccgcttt tcagctcgcc gatccggtcc
     1081 agcaggtact ggttggtgct gtcgatcacg ggcagcacag ccacagatcc gccatccagc
     1141 tggcccagga tctgcttggc gttcagcagc tggataggct cgggcaggct gtagcccttg
     1201 ccaggcacgg tgaacacgtc cacgccccag tctctcaggg tctggatgtg cttgttgatg
     1261 gcggctctag acatgcccag tgtctcgccc agctgctcgc cagagtggaa ctcgccattg
     1321 gccagcaggg cgatcagctt caggggcacg gtgttgtcct tggatccttt atcgtcatcg
     1381 tctttgtagt ccatggtggc tgatccgagc tcggtaccaa gcttgggtct ccctatagtg
     1441 agtcgtatta atttcgataa gccagtaagc agtgggttct ctagttagcc agagagctct
     1501 gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt
     1561 gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc
     1621 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
     1681 ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt
     1741 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     1801 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     1861 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     1921 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     1981 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
     2041 aattgattac tattaataac tagtcaataa tcaatgtcaa cgcgtatatc tggcccgtac
     2101 atcgcgaagc agcgcaaaac gcctaaccct aagcagattc ttcatgcaat tgtcggtcaa
     2161 gccttgcctt gttgtagctt aaattttgct cgcgcactac tcagcgacct ccaacacaca
     2221 agcagggagc agatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     2281 ccatagggga tcgggagatc cgatgcatgg ggtcgtgcgc tcctttcggt cgggcgctgc
     2341 gggtcgtggg gcgggcgtca ggcaccgggc ttgcgggtca tgcaccaggt gcgcggtcct
     2401 tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc gctcgtagaa ggggaggttg
     2461 cggggcgcgg aggtctccag gaaggcgggc accccggcgc gctcggccgc ctccactccg
     2521 gggagcacga cggcgctgcc cagacccttg ccctggtggt cgggcgagac gccgacggtg
     2581 gccaggaacc acgcgggctc cttgggccgg tgcggcgcca ggaggccttc catctgttgc
     2641 tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg ggccgatctc ggcgaacacc
     2701 gcccccgctt cgacgctctc cggcgtggtc cagaccgcca ccgcggcgcc gtcgtccgcg
     2761 acccacacct tgccgatgtc gagcccgacg cgcgtgagga agagttcttg cagctcggtg
     2821 acccgctcga tgtggcggtc cggatcgacg gtgtggcgcg tggcggggta gtcggcgaac
     2881 gcggcggcga gggtgcgtac ggccctgggg acgtcgtcgc gggtggcgag gcgcaccgtg
     2941 ggcttgtact cggtcatggt aagctgatcc ggccggcgcc tagagaagga gtgagggctg
     3001 gataaaggga ggatcgaggc ggggtcgaac gaggaggttc aagggggaga gacggggcgg
     3061 atggaggaag aggaggcgga ggcttagggt gtacaaaggg cttgacccag ggaggggggt
     3121 caaaagccaa ggcttcccag gtcacgatgt aggggacctg gtctgggtgt ccatgcgggc
     3181 caggtgaaaa gaccttgatc ttaacctggg tgatgaggtc tcggttaaag gtgccgtctc
     3241 gcggccatcc gacgttaaag gttggccatt ctgcagagca gaaggtaacc caacgtctct
     3301 tcttgacatc taccgactgg ttgtgagcga tccgctcgac atctttccag tgacctaagg
     3361 tcaaacttaa gggagtggta acagtctggc cctaattttc agacaaatac agaaacacag
     3421 tcagacagag acaacacaga acgatgctgc agcgctgcag cagacaagac gcgcggcttc
     3481 ggttccaaac cgaaagcaaa aattcagacg gaggcgggaa ctgttttagg ttctcgtctc
     3541 ctaccagaac cacatatcct gacggggtcg gattccacat cgactccctt cctcaggtcg
     3601 ggccacaaaa acggccccca aagtccctgg gacgtctccc agggttgcgg ccgggtgttc
     3661 agaactcgtc agttccacca cgggtccgcc agatacagag ctagttagct aactagtacc
     3721 gacgcaggcg cataaaatca gtcatagaca ctagacaatc ggacagacac agataagttg
     3781 ctggccagct tgcctcccgg tggtgggtcg gtggtccctg ggcaggggtc tcccgatccc
     3841 ggacgagccc ccaaatgaaa gacccccgct gacgggtagt caatcactca gaggagaccc
     3901 tcccaaggaa cagcgagacc acaagtcgga tgcaactgca agagggttta ttggatacac
     3961 gggtacccgg gcgactcagt caatcggagg actggcgccc cgagtgaggg gttgtgggct
     4021 cttttattga gctcggggag cagaagcgcg cgaacagaag cgagaagcga actgattggt
     4081 tagttcaaat aaggcacagg gtcatttcag gtccttgggg caccctggaa acatctgatg
     4141 gttctctaga aactgctgag ggctggaccg catctgggga ccatctgttc ttggccctga
     4201 gccggggcag gaactgctta ccacagatat cctgtttggc ccatattcag ctgttccatc
     4261 tgttcttggc cctgagccgg ggcaggaact gcttaccaca gatatcctgt ttggcccata
     4321 ttcaggctgc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4381 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4441 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4501 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4561 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4621 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4681 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4741 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4801 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4861 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4921 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4981 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5041 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5101 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5161 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5221 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5281 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5341 catatatact ttagattgat ttgcggccgc aaaaggatct aggtgaagat cctttttgat
     5401 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5461 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5521 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5581 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5641 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5701 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5761 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5821 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5881 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5941 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6001 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6061 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6121 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6181 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6241 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6301 tgcaactatg gccatttaat gtaaatactt aagaaaaaaa accaaattaa ttttgataca
     6361 tgctgcatgt gaaagacccc cgctgacggg tagtcaatca ctcagaggag accctcccaa
     6421 ggaacagcga gaccacaagt cggatgcaac tgcaagaggg tttattggat acacgggtac
     6481 ccgggcgact cagtcaatcg gaggactggc gccccgagtg aggggttgtg ggctctttta
     6541 ttgagctcgg ggagcagaag cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc
     6601 aaataaggca cagggtcatt tcaggtcctt ggggcaccct ggaaacatct gatggttctc
     6661 tagaaactgc tgagggctgg accgcatctg gggaccatct gttcttggcc ctgagccggg
     6721 gcaggaactg cttaccacag atatcctgtt tggcccatat tcagctgttc catctgttct
     6781 tggccctgag ccggggcagg aactgcttac cacagatatc ctgtttggcc catattcagc
     6841 tgttccatct gttcctgacc ttgatctgaa cttctctatt ctcagttatg tatttttcca
     6901 tgccttgcaa aatggcgtta cttaagctag cttgccaaac ctacaggtgg ggtctttcat
     6961 tccccccttt ttctggagac taaataaaat cttttatttt atcggccctc tagatgcatg
     7021 cttatttctt tcggaattca atcctccgag caacgttctg agctccaaac tttttgacca
     7081 gtgcattttt cgtgtcgtca tcgtccttct gcaaatctaa gtcatcttgt ctgcaccaaa
     7141 tgggataacc gtcgggtctc cgccccgact ctaagaagga ggacgtggcc tccagctcac
     7201 cactgttctt cagcaaggcc tgtgtaactg ttgatagatc caagttgaac ttctccatta
     7261 gctgccggcg caccttaatg gcagttccca cttcttgcgt agaaactttt ggttcttctt
     7321 cctcctcgtc tggctgtgtt tctgagtctt cctcgggtgt gggtggatca ccatcacaca
     7381 tcgttatatg tatttcaaag tcagggctct catccaccac ggcttctccg aactccggag
     7441 cggcttcctc aaacagcttg ttgtctgcct ctccattctc cttgatctct cctttcacac
     7501 actcctcctc aggcaagtct ggcgctctct tgttctgtgg ctctccgcta tccgcggcct
     7561 cggggtcctg ctccgccttc cgtttgagct tctgggagga cgggctgacc ggggcgttcc
     7621 cgagcaggta cttgtgctcc tgaccccgta ggtgcttgag gtagcggtcc ttgagcgact
     7681 gccaggagtg ctgcgtgagc gagctcttct ccatcgcttt ccacaaggca ttgcctgtga
     7741 ccgagctggg cgaacgggcg ttctccttca cgtaggtcag gatggccaca tcttccgcgt
     7801 cggtgtaggc gatgcgcccg gtcaggggct gcggctccgg ttccgtggag ccctcggtgg
     7861 aagccccggg cttcggatcg gacgcctgct ccgtcaggcc cagccgatag gcctccaggt
     7921 ccagcttctc gttgcgatcc acgcagtct
//
