LOCUS       sequence-242765-        6880 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the restriction enzyme under the control of Shield1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6880)
  AUTHORS   Goldstein M, Derheimer FA, Tait-Mulder J, Kastan MB
  TITLE     Nucleolin mediates nucleosome disruption critical for DNA 
            double-strand break repair.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Sep 30.
  PUBMED    24082117
REFERENCE   2  (bases 1 to 6880)
  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: "Proc Natl 
            Acad Sci U S A. 2013 Sep 30."
COMMENT     Sequence Label: pBABE-ddIPpoI
FEATURES             Location/Qualifiers
     source          1..6880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..67)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        52..381
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      232..367
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             391..990
                     /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(391..410)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     887..907
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1211..1680
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(1810..1827)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1981..2569)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2061..2080)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2729..3520,3521..3589))
                     /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     3352..3371
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3590..3694)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             3725..4194
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    4259..4458
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             4659..5075
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5005..5027
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5043..5059
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             join(5086..5088,5089..5409)
                     /codon_start=1
                     /product="destabilization domain that can be stabilized by 
                     Shield1 in the ProteoTuner(TM) system"
                     /label=DD
                     /note="L106P mutant of FKBP12"
                     /translation="MGVQVQTISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKPE"
     CDS             5425..5451
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 gtcctcaaca aaggggcagt cactttgtgg tataagtcga ccctgtggaa tgtgtgtcag
       61 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
      121 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
      181 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
      241 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
      301 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
      361 ggaggcctag gcttttgcaa aaagcttacc atgaccgagt acaagcccac ggtgcgcctc
      421 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
      481 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
      541 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
      601 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
      661 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
      721 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
      781 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
      841 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
      901 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
      961 tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc acgacccgca gcgcccgacc
     1021 gaaaggagcg cacgacccca tgcatcgata aaataaaaga ttttatttag tctccagaaa
     1081 aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt
     1141 tgcaaggcat ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca
     1201 gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
     1261 tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc
     1321 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag
     1381 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt
     1441 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc
     1501 cgagctcaat aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga
     1561 gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct
     1621 cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca
     1681 catgcagcat gtatcaaaat taatttggtt ttttttctta agtatttaca ttaaatggcc
     1741 atagttgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1801 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     1861 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     1921 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1981 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2041 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2101 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2161 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2221 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2281 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2341 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2401 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2461 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2521 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2581 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2641 catgagatta tcaaaaagga tcttcaccta gatccttttg cggccgcaaa tcaatctaaa
     2701 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2761 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2821 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     2881 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2941 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3001 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3061 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3121 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3181 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3241 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3301 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3361 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3421 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3481 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3541 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3601 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3661 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     3721 cagcctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     3781 ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc
     3841 tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc tcagcagttt
     3901 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     3961 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     4021 caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc
     4081 cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt
     4141 tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatttggg
     4201 ggctcgtccg ggatcgggag acccctgccc agggaccacc gacccaccac cgggaggcaa
     4261 gctggccagc aacttatctg tgtctgtccg attgtctagt gtctatgact gattttatgc
     4321 gcctgcgtcg gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg
     4381 acgagttctg aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt
     4441 tttgtggccc gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg
     4501 ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg
     4561 tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc
     4621 tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct
     4681 taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag
     4741 atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg
     4801 gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct
     4861 tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct
     4921 tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc
     4981 ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct
     5041 ccctttatcc agccctcact ccttctctag gcgccggccg gatccatggg agtgcaggtg
     5101 cagactatct ccccaggaga cgggcgcacc ttccccaagc gcggccagac ctgcgtggtg
     5161 cactacaccg ggatgcttga agatggaaag aaagttgatt cctcccggga cagaaacaag
     5221 ccctttaagt ttatgctagg caagcaggag gtgatccgag gctgggaaga aggggttgcc
     5281 cagatgagtg tgggtcagag agccaaactg actatatctc cagattatgc ctatggtgcc
     5341 actgggcacc caggcatcat cccaccacat gccactctcg tcttcgatgt ggagcttcta
     5401 aaaccggaag aattcaccat ggcataccca tacgatgttc cagattacgc tagcggatct
     5461 cgaaatgaaa tgggtgcttc aggagacatg agggctgcca acctttggcc aagccctctt
     5521 gtgattaagc acactaagaa gaatagccct gccttgtcct tgacagctga ccagatggtc
     5581 agtgccttgt tggatgctga accgcccatg atctattctg aatatgatcc ttctagaccc
     5641 ttcagtgaag cctcaatgat gggcttattg accaacctag cagataggga gctggttcat
     5701 atgatcaact gggcaaagag agtgccaggc tttggggact tgaatctcca tgatcaggtc
     5761 caccttctcg agtgtgcctg gctggagatt ctgatgattg gtctcgtctg gcgctccatg
     5821 gaacacccgg ggaagctcct gtttgctcct aacttgctcc tggacaggaa tcaaggtaaa
     5881 tgtgtggaag gcatggtgga gatctttgac atgttgcttg ctacgtcaag tcggttccgc
     5941 atgatgaacc tgcagggtga agagtttgtg tgcctcaaat ccatcatttt gcttaattcc
     6001 ggagtgtaca cgtttctgtc cagcaccttg aagtctctgg aagagaagga ccacatccac
     6061 cgtgtcctgg acaagatcac agacactttg atccacctga tggccaaagc tggcctgact
     6121 ctgcagcagc agcatcgccg cctagctcag ctccttctca ttctttccca tatccggcat
     6181 atgagtaaca aacgcatgga gcatctctac aacatgaaat gcaagaacgt ggtacccctc
     6241 tatgacctgc tcctggagat gttggatgcc caccgccttc atgccccagc cagtcgcatg
     6301 ggagtgcccc cagaggagcc cagccagacc cagctggcca ccaccagctc cacttcagca
     6361 cattccttac aaacctacta catacccccg gaagcagagg gcttccccaa cacgatcgga
     6421 tctatggcgc tcaccaatgc tcaaatcttg gctgtgattg acagctggga agaaacagtc
     6481 ggtcagtttc cagtgataac gcaccatgta ccattaggtg gcggtctgca aggaacgctc
     6541 cattgttacg agatccccct agcagctcct tatggggttg gctttgctaa gaatgggcct
     6601 acccgctggc aatacaaacg gacaatcaat caagtcgtcc acagatgggg atcccacaca
     6661 gtcccttttc tattagaacc ggataacatc aacggcaaaa cctgcacagc atcgcaccta
     6721 tgtcataata ctcgatgcca caatcccttg cacttgtgct gggagtcact agacgacaac
     6781 aaaggcagaa actggtgccc gggtcccaac gggggatgtg tccatgcggt ggtttgttta
     6841 aggcagggtc cgttgtacgg cccgggggcg actgtggcag
//
