LOCUS       sequence-301991-        9649 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For transient expression of mouse PDS5B tagged with mKate2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9649)
  AUTHORS   Nora EP, Caccianini L, Fudenberg G, So K, Kameswaran V, Nagle A, 
            Uebersohn A, Hajj B, Saux AL, Coulon A, Mirny LA, Pollard KS, Dahan 
            M, Bruneau BG
  TITLE     Molecular basis of CTCF binding polarity in genome folding.
  JOURNAL   Nat Commun. 2020 Nov 5;11(1):5612. doi: 10.1038/s41467-020-19283-x.
  PUBMED    33154377
REFERENCE   2  (bases 1 to 9649)
  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: "Nat 
            Commun."; date: "2020-11-5"; volume: "11(1):5612. doi"; pages: " 
            10.1038/s41467-020-19283-x"
COMMENT     Sequence Label: pKS023 - pCAGGS-3XFLAG-mKate2-Pds5b-bpA
FEATURES             Location/Qualifiers
     source          1..9649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1757..1822
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1844..2539
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     CDS             6472..6483
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7010..7029)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7075..7130
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7129..7148)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7489..7505)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7489..7505)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7502..7524)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7513..7529
                     /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(7537..7543,7544..7561,7562..7567))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7582..7603
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7728..7745)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7899..8487)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7979..7998)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8658..9449,9450..9518))
                     /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     9281..9300
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9519..9623)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcctcgag aattctcacg
     1741 cgtgaattcc accatggact ataaggacca cgacggagac tacaaggatc atgatattga
     1801 ttacaaagac gatgacgata agcccggggg gggagcagga ggcatggtga gcgagctgat
     1861 taaggagaac atgcacatga agctgtacat ggagggcacc gtgaacaacc accacttcaa
     1921 gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggc
     1981 ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctaccagct tcatgtacgg
     2041 cagcaaaacc ttcatcaacc acacccaggg catccccgac ttctttaagc agtccttccc
     2101 cgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac
     2161 ccaggacacc agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa
     2221 cttcccatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg cctccaccga
     2281 gaccctgtac cccgctgacg gcggcctgga aggcagagcc gacatggccc tgaagctcgt
     2341 gggcgggggc cacctgatct gcaacttgaa gaccacatac agatccaaga aacccgctaa
     2401 gaacctcaag atgcccggcg tctactatgt ggacagaaga ctggaaagaa tcaaggaggc
     2461 cgacaaagag acctacgtcg agcagcacga ggtggctgtg gccagatact gcgacctccc
     2521 tagcaaactg gggcacagag gcggtggagg gtcaggcggt ggtggatcca ccggtatggc
     2581 tcattcaaag acaaggacca acgatgggaa aattacttac cctcctggag tcaaggaaat
     2641 ctcagataaa atctctaaag aggagatggt gaggcggtta aagatggttg taaaaacttt
     2701 catggacatg gaccaggact ctgaagagga aaaggaactt tatctaaacc tagctttaca
     2761 tcttgcttct gacttcttcc tcaagcatcc tgataaagat gttcgtttac tggtggcttg
     2821 ctgccttgct gatattttca ggatttatgc tcctgaggct ccttacacgt ctcccgataa
     2881 actaaaggat atatttatgt ttataacaag gcaactaaag ggactagaag atacaaagag
     2941 ccctcaattt aataggtatt tttatttact tgagaacatt gcatgggtta aatcatataa
     3001 catatgcttt gagttagaag atagcaatga aatctttact caactttaca gaacattatt
     3061 ctcagttata aacaatggcc acaatcagaa agttcatatg cacatggtgg acctcatgag
     3121 ctctatcatt tgtgaaggtg ataccgtatc tcaggagctc ttagatacag ttttagtaaa
     3181 tctggtacct gcccataaga acttaaacaa gcaagcatat gatttggcaa aggctttgct
     3241 gaagaggact gctcaagcta ttgaaccata tattaccaat ttttttaacc aggttctgat
     3301 gcttgggaaa acatctatca gtgatttgtc tgagcatgtc tttgatttga tcttggaact
     3361 ctacaacatt gacagccatt tgctgctttc tgttctgccc cagcttgagt ttaaattaaa
     3421 gagcaatgat aatgaagaac gcctccaggt tgttaaacta ctggcgaaaa tgtttggggc
     3481 aaaggattca gaattggctt ctcagaacaa acccctctgg cagtgctacc tgggcaggtt
     3541 taatgatatc catgtaccaa tacgcctgga atgtgtgaaa tttgccagcc actgcctcat
     3601 gaaccatcct gatttagcaa aggatttaac agagtatctt aaagtgaggt cacatgatcc
     3661 tgaggaagct attagacatg atgttattgt gtctatagta acagctgcta aaaaggatat
     3721 tcttcttgtc aatgatcact tactcaattt tgtgagagag cgaacattgg acaaacggtg
     3781 gagagtacgt aaagaagcca tgatgggact ggcccagatt tataagaaat actctttaca
     3841 gtcagcagct ggcaaagatg ctgcaaaaca gatatcctgg gtcaaggaca agctgctgca
     3901 tatatactat cagaacagca tcgacgaccg gctacttgtt gagcggatct ttgctcagta
     3961 catggtccct cacaatttgg aaacgacgga acggatgaag tgcttgtatt acttgtacgc
     4021 tacactggac ttgaatgctg tgaaagcatt gaatgaaatg tggaaatgtc aaaatctact
     4081 ccgacatcaa gtaaaggatt tacttgattt aattaaacaa ccaaaaacag atgccagtgt
     4141 caaggccata ttttctaaag tgatggttat cacaagaaat ttgccagatc ctggtaaggc
     4201 ccaagacttc atgaagaagt tcacacaagt gttggaagat gatgagaaaa tacggaaaca
     4261 gctggaagcc cttgttagtc ccacatgttc ctgcaagcag gctgaaggct gtgtgcgtga
     4321 aataactaag aagttgggca accctaagca gcctacaaac cctttcttgg aaatgatcaa
     4381 gtttctcttg gagaggatag ctcctgtaca catagacact gaatccatca gtgctcttat
     4441 caagcaagtg aataaatcaa tagatggaac agcagatgat gaggatgagg gtgttccaac
     4501 tgatcaagct atcagggcag gccttgagct gctgaaggtg ctgtctttta cgcaccctat
     4561 ctcctttcac tctgctgaga cgtttgagtc ccttctggct tgtctgaaga tggatgatga
     4621 gaaggtggca gaagctgcac tacaaatttt caaaaacaca ggaagcaaaa ttgaagagga
     4681 tttccctcac atcagatcag ccttgcttcc tgtactacat cacaaatcca aaaaaggacc
     4741 acctcggcag gccaaatacg ccatccattg tattcatgcc atattttcta gtaaagaaac
     4801 ccagtttgca cagatatttg agcctctgca taaaagccta gatccaagca acctggagca
     4861 tcttataacc cctctggtca ctattggtca tattgctctt ctcgcacctg atcagttcgc
     4921 tgctcctctg aagtctttgg tggcaacttt cattgtgaag gacctcctca tgaatgaccg
     4981 gcttccagga aaaaagacaa ctaagctttg ggttccagat gaggaagtct cacctgagac
     5041 aatggtcaaa attcaggcta ttaaaatgat ggttcgatgg ctacttggaa tgaagaataa
     5101 tcacagtaag tcaggaactt ccaccctcag attactaaca acgatactgc atagtgatgg
     5161 ggatttgaca gaacaaggga aaattagtaa accagatatg tcacgcctga gacttgctgc
     5221 tgggagtgct attgtgaagc tggcacagga gccctgttac cacgagatca ttacactgga
     5281 gcagtaccag ctgtgtgcat tagccatcaa tgatgagtgt tatcaagtca ggcaggtgtt
     5341 cgctcagaaa cttcacaaag gcctttcccg cttacggctt ccccttgagt acatggccat
     5401 ctgtgctctt tgtgccaaag accctgtgaa agagaggcga gcccatgcta gacagtgctt
     5461 ggtgaagaac atcactgtga ggagggagta cctgaagcag catgcagctg ttagtgaaaa
     5521 attattgtct cttctaccag agtatgttgt tccatataca attcaccttt tggcacatga
     5581 cccagattat gtcaaagtac aggatattga acaacttaaa gatgtgaaag aatgcctttg
     5641 gtttgttctg gagatattga tggctaaaaa tgaaaacaac agccatgcat ttatcagaaa
     5701 aatggtagaa aatattaaac agacaaaaga tgctcaagga ccagatgata caaaaatgaa
     5761 tgaaaaattg tacaccgtgt gtgatgttgc catgaacatc atcatgtcca agagcaccac
     5821 gtacagcctg gagtctccta aggaccccgt gctgccagct cggtttttca cccagcctga
     5881 caagaatttt agtaacacca aaaattacct gcctccagaa atgaaatcat ttttcactcc
     5941 tggaaaacct aaaacagcca atgttctcgg agcagttaat aagccacttt catcagcagg
     6001 caaacagtct cagaccaaat catcaagaat ggaaactgtg agcaacgcaa gcagcagctc
     6061 caacccaagc tctcctggaa ggatcaaggg gaggcttgat agctctgaaa tggatcacag
     6121 tgaaaatgaa gattatacaa tgtcttcacc tttgccagga aaaaaaagtg acaagagaga
     6181 agaccctgat ctttctgagt tggagaagcc tagaagtcgg aaaaaagcac ctgtcacaga
     6241 ccctgaagag aaattaggta tggatgacct aactaagttg gtacaggaac agaaacctaa
     6301 aggcagtcag cgaggccgga aaagaggccg tacagcttca gactcagacg agcagcagtg
     6361 gcctgaggag aagaggcaca aagaggagct cctggaaaat gaggatgagc agaacagccc
     6421 accaaaaaag ggcaaaagag gcaggccacc aaaacctctt ggtgggggga catcgaagga
     6481 agagcccaca atgaaaacat ccaagaaagg aaacaagaaa aaacttgtac ctcctgtagt
     6541 agacgacgat gaagaagaag aaagacaaat tggaaacaca gaacataagt caaaaagcaa
     6601 acagcaccgc acatcaaaga gagcacaaca gagagcagaa tctcctgaaa caagtgcagt
     6661 tgaatccaca cagtccacac cacagaaagg acgaggaaga ccatcaaaag caccatcacc
     6721 atcacaaccc cccaaaaaaa tccgtgtagg acgctccaaa caggtagcca ctaaagaaaa
     6781 cgattcaagt gaagaaatgg atgtgcttca ggccagctct cctgtcagtg acgacactac
     6841 acaggaagga gcagaagagg aggacatttc tgtgggaaat gtacgacggc gaagctccaa
     6901 acgagagaga cgatgagttt aaacgcggcc gctcaggtca ggtgcaggct gcctatcaga
     6961 aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct
     7021 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa
     7081 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     7141 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
     7201 catatgccat atgctggctg ccatgaacaa aggtggctat aaagaggtca tcagtatatg
     7261 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
     7321 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
     7381 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
     7441 ctcttatgaa gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt
     7501 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     7561 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     7621 gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgctgcatta atgaatcggc
     7681 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     7741 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     7801 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     7861 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7921 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7981 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     8041 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     8101 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     8161 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     8221 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     8281 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     8341 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     8401 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     8461 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     8521 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     8581 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8641 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8701 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8761 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8821 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8881 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8941 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     9001 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     9061 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     9121 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     9181 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     9241 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     9301 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     9361 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     9421 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9481 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9541 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9601 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctg
//
