LOCUS       sequence-163147-        8796 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expession of FLAG-PALB2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8796)
  AUTHORS   Orthwein A, Noordermeer SM, Wilson MD, Landry S, Enchev RI, Sherker 
            A, Munro M, Pinder J, Salsman J, Dellaire G, Xia B, Peter M, 
            Durocher D
  TITLE     A mechanism for the suppression of homologous recombination in G1 
            cells.
  JOURNAL   Nature. 2015 Dec 9. doi: 10.1038/nature16142.
  PUBMED    26649820
REFERENCE   2  (bases 1 to 8796)
  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: "Nature. 
            2015 Dec 9. doi: 10.1038/nature16142."
COMMENT     Sequence Label: pDEST-FRT/T0-Flag-PALB2
FEATURES             Location/Qualifiers
     source          1..8796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1034..1057
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    1058..1082
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4648..4672)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4672..4696)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4748..4765)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4754..4978
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5111..5130)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5262..5309
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             5317..6339
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGDFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    6469..6590
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6506..6525)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6560..6579
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6639..6655)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6639..6655)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6652..6674)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6663..6679
                     /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(6687..6693,6694..6711,6712..6717))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6732..6753
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6870..6887)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7041..7629)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7121..7140)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7800..8591,8592..8660))
                     /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     8423..8442
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8661..8765)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttccaccat ggactacaaa gaccatgacg gtgatacata
     1021 tgatcatgac atcgattaca aggatgacga tgacaagaca agtttgtaca aaaaagcagg
     1081 ctccaccatg gacgagcctc ccgggaagcc cctcagctgt gaggagaagg aaaagttaaa
     1141 ggagaaatta gcattcttga aaagggaata cagcaagaca ctagcccgcc ttcagcgtgc
     1201 ccaaagagct gaaaagatta agcattctat taagaaaaca gtagaagaac aagattgttt
     1261 gtctcagcag gatctctcac cgcagctaaa acactcagaa cctaaaaata aaatatgtgt
     1321 ttatgacaag ttacacatca aaacccatct tgatgaagaa actggagaaa agacatctat
     1381 cacacttgat gttgggcctg agtcctttaa ccctggagat ggcccaggag gattacctat
     1441 acaaagaaca gatgacaccc aagaacattt tccccacagg gtcagtgacc ctagtggtga
     1501 gcaaaagcag aagctgccaa gcagaagaaa gaagcagcag aagaggacat ttatttcaca
     1561 ggagagagac tgtgtctttg gcactgattc actcagattg tctgggaaaa gactaaagga
     1621 acaggaagaa atcagtagca aaaatcctgc tagatcacca gtaactgaaa taagaactca
     1681 ccttttaagt cttaaatctg aacttccaga ttctccagaa ccagttacag aaattaatga
     1741 agacagtgta ttaattccac caactgccca accagaaaaa ggtgttgata cattcctaag
     1801 aagacctaat ttcaccaggg cgactacagt tcctttacag actctatcag atagcggtag
     1861 tagtcagcac cttgaacaca ttcctcctaa aggtagcagt gaacttacta ctcacgacct
     1921 aaaaaacatt agatttactt cacctgtaag tttggaggca caaggcaaaa aaatgactgt
     1981 ctctacagat aacctccttg taaataaagc tataagtaaa agtggccaac tgcccacaag
     2041 ttctaattta gaggcaaata tttcatgttc tctaaatgaa ctcacctaca ataacttacc
     2101 agcaaatgaa aaccaaaact taaaagaaca aaatcaaaca gagaaatctt taaaatctcc
     2161 cagtgacact cttgatggca ggaatgaaaa tcttcaggaa agtgagattc taagtcaacc
     2221 taagagtctt agcctggaag caacctctcc tctttctgca gaaaaacatt cttgcacagt
     2281 gcctgaaggc cttctgtttc ctgcagaata ttatgttaga acaacacgaa gcatgtccaa
     2341 ttgccagagg aaagtagccg tggaggctgt cattcagagt catttggatg tcaagaaaaa
     2401 agggtttaaa aataaaaata aggatgcaag taaaaattta aacctttcca atgaggaaac
     2461 tgaccaaagt gaaattagga tgtctggcac atgcacagga caaccaagtt caagaacctc
     2521 tcagaaactt ctctcattaa ctaaagtcag ctctcccgct gggcccactg aagataatga
     2581 cttgtctagg aaggcagttg cccaagcacc tggtagaaga tacacaggaa aaagaaaatc
     2641 agcctgcacc ccagcatcag atcattgtga accacttttg ccaacttcta gcctgtcgat
     2701 tgttaacagg tccaaggaag aagtcacctc acacaaatat cagcacgaaa aattatttat
     2761 tcaagtgaaa gggaagaaaa gtcgtcatca aaaagaggat tccctttctt ggagtaatag
     2821 tgcttattta tccttggatg atgatgcttt cacggctcca tttcataggg atggaatgct
     2881 gagtttaaag caactactgt cttttctcag tatcacagac tttcagttac ctgatgaaga
     2941 ctttggacct cttaagcttg aaaaagtgaa gtcctgctca gaaaaaccag tggagccctt
     3001 tgagtcaaaa atgtttggag agagacatct taaagaggga agctgtattt ttccagagga
     3061 actgagtcct aaacgcatgg atacagaaat ggaggactta gaagaggacc ttattgttct
     3121 accaggaaaa tcacatccca aaaggccaaa ctcgcaaagc cagcatacaa agacgggcct
     3181 ttcttcatcc atattacttt atactccttt aaatacggtt gcgcctgatg ataatgacag
     3241 gcctaccaca gacatgtgtt cacctgcttt ccccatctta ggtactactc cagcctttgg
     3301 ccctcaaggc tcctatgaaa aagcatctac agaagttgct ggacgaactt gctgcacacc
     3361 ccaacttgct catttgaaag actcagtctg tcttgccagt gatactaaac aattcgacag
     3421 ttcaggcagc ccagcaaaac cacataccac cctgcaagtg tcaggcaggc aaggacaacc
     3481 tacctgtgac tgtgactctg tcccgccagg aacacctcca cccattgagt cattcacttt
     3541 taaagaaaat cagctctgta gaaacacatg ccaggagctg cataaacatt ccgtcgaaca
     3601 gactgaaaca gcagagcttc ctgcttctga tagcataaac ccaggcaacc tacaattggt
     3661 ttcagagtta aagaatcctt caggttcctg ttccgtagat gtgagtgcca tgttttggga
     3721 aagagccggt tgtaaagagc catgtatcat aactgcttgc gaagatgtag tttctctttg
     3781 gaaagctctg gatgcttggc agtgggaaaa actttatacc tggcacttcg cagaggttcc
     3841 agtattacag atagttccag tgcctgatgt gtataatctc gtgtgtgtag ctttgggaaa
     3901 tttggaaatc agagagatca gggcattgtt ttgttcctct gatgatgaaa gtgaaaagca
     3961 agtactactg aagtctggaa atataaaagc tgtgcttggc ctgacaaaga ggaggctagt
     4021 tagtagcagt gggacccttt ctgatcaaca agtagaagtc atgacgtttg cagaagatgg
     4081 aggaggcaaa gaaaaccaat ttttgatgcc ccctgaggag actatactaa cttttgctga
     4141 ggtccaaggg atgcaagaag ctctgcttgg tactactatt atgaacaaca ttgttatttg
     4201 gaatttaaaa actggtcaac tcctgaaaaa gatgcacatt gatgattctt accaagcttc
     4261 agtctgtcac aaagcctatt ctgaaatggg gcttctcttt attgtcctga gtcatccctg
     4321 tgccaaagag agtgagtcgt tgcgaagccc tgtgtttcag ctcattgtga ttaaccctaa
     4381 gacgactctc agcgtgggtg tgatgctgta ctgtcttcct ccagggcagg ctggcaggtt
     4441 cctggaaggt gacgtgaaag atcactgtgc agcagcaatc ttgacttctg gaacaattgc
     4501 catttgggac ttacttctcg gtcagtgtac tgccctcctc ccacctgtct ctgaccaaca
     4561 ttggtctttt gtgaaatggt cgggtacaga ctctcatttg ctggctggac aaaaagatgg
     4621 aaatatattt gtataccact attcataaac ccgctttctt gtacaaagtg gtttcagctt
     4681 tcttgtacaa agtggtgtga ctcgagctga ctcgagtcta gagggcccgt ttaaacccgc
     4741 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     4801 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     4861 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     4921 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     4981 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     5041 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5101 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5161 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     5221 gaccccaaaa aacttgatta gggtgatggt tcacgtacct agaagttcct attccgaagt
     5281 tcctattctc tagaaagtat aggaacttcc ttggccaaaa agcctgaact caccgcgacg
     5341 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg
     5401 gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg
     5461 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg
     5521 gccgcgctcc cgattccgga agtgcttgac attggggatt tcagcgagag cctgacctat
     5581 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc
     5641 gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag
     5701 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat
     5761 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
     5821 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc
     5881 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc
     5941 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc
     6001 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc
     6061 gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt
     6121 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg
     6181 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc
     6241 gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga
     6301 aaccgacgcc ccagcactcg tccgagggca aaggaatagc acgtactacg agatttcgat
     6361 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     6421 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     6481 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6541 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     6601 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     6661 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6721 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6781 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     6841 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6901 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6961 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     7021 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     7081 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     7141 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     7201 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     7261 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     7321 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     7381 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     7441 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     7501 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     7561 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7621 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7681 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7741 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7801 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7861 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7921 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7981 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     8041 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     8101 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     8161 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     8221 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     8281 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     8341 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     8401 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     8461 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     8521 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     8581 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     8641 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8701 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8761 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
