LOCUS       sequence-310174-        9659 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 9659)
  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 9659)
  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..9659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1765..1830
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1852..2547
                     /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             6480..6491
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7018..7037)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7083..7138
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7137..7156)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7499..7515)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7499..7515)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7512..7534)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7523..7539
                     /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(7547..7553,7554..7571,7572..7577))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7592..7613
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7738..7755)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7909..8497)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7989..8008)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8668..9459,9460..9528))
                     /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     9291..9310
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9529..9633)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggsgcgc gccrggsggg ggggggsggg gsgrggggsg
      541 gggsggggsg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcctcgagaa
     1741 ttctcacgcg tgaattccac catggactat aaggaccacg acggagacta caaggatcat
     1801 gatattgatt acaaagacga tgacgataag cccggggggg gagcaggagg catggtgagc
     1861 gagctgatta aggagaacat gcacatgaag ctgtacatgg agggcaccgt gaacaaccac
     1921 cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga
     1981 atcaaggcgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc taccagcttc
     2041 atgtacggca gcaaaacctt catcaaccac acccagggca tccccgactt ctttaagcag
     2101 tccttccccg agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg
     2161 accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga
     2221 ggggtgaact tcccatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc
     2281 tccaccgaga ccctgtaccc cgctgacggc ggcctggaag gcagagccga catggccctg
     2341 aagctcgtgg gcgggggcca cctgatctgc aacttgaaga ccacatacag atccaagaaa
     2401 cccgctaaga acctcaagat gcccggcgtc tactatgtgg acagaagact ggaaagaatc
     2461 aaggaggccg acaaagagac ctacgtcgag cagcacgagg tggctgtggc cagatactgc
     2521 gacctcccta gcaaactggg gcacagaggc ggtggagggt caggcggtgg tggatccacc
     2581 ggtatggctc attcaaagac aaggaccaac gatgggaaaa ttacttaccc tcctggagtc
     2641 aaggaaatct cagataaaat ctctaaagag gagatggtga ggcggttaaa gatggttgta
     2701 aaaactttca tggacatgga ccaggactct gaagaggaaa aggaacttta tctaaaccta
     2761 gctttacatc ttgcttctga cttcttcctc aagcatcctg ataaagatgt tcgtttactg
     2821 gtggcttgct gccttgctga tattttcagg atttatgctc ctgaggctcc ttacacgtct
     2881 cccgataaac taaaggatat atttatgttt ataacaaggc aactaaaggg actagaagat
     2941 acaaagagcc ctcaatttaa taggtatttt tatttacttg agaacattgc atgggttaaa
     3001 tcatataaca tatgctttga gttagaagat agcaatgaaa tctttactca actttacaga
     3061 acattattct cagttataaa caatggccac aatcagaaag ttcatatgca catggtggac
     3121 ctcatgagct ctatcatttg tgaaggtgat accgtatctc aggagctctt agatacagtt
     3181 ttagtaaatc tggtacctgc ccataagaac ttaaacaagc aagcatatga tttggcaaag
     3241 gctttgctga agaggactgc tcaagctatt gaaccatata ttaccaattt ttttaaccag
     3301 gttctgatgc ttgggaaaac atctatcagt gatttgtctg agcatgtctt tgatttgatc
     3361 ttggaactct acaacattga cagccatttg ctgctttctg ttctgcccca gcttgagttt
     3421 aaattaaaga gcaatgataa tgaagaacgc ctccaggttg ttaaactact ggcgaaaatg
     3481 tttggggcaa aggattcaga attggcttct cagaacaaac ccctctggca gtgctacctg
     3541 ggcaggttta atgatatcca tgtaccaata cgcctggaat gtgtgaaatt tgccagccac
     3601 tgcctcatga accatcctga tttagcaaag gatttaacag agtatcttaa agtgaggtca
     3661 catgatcctg aggaagctat tagacatgat gttattgtgt ctatagtaac agctgctaaa
     3721 aaggatattc ttcttgtcaa tgatcactta ctcaattttg tgagagagcg aacattggac
     3781 aaacggtgga gagtacgtaa agaagccatg atgggactgg cccagattta taagaaatac
     3841 tctttacagt cagcagctgg caaagatgct gcaaaacaga tatcctgggt caaggacaag
     3901 ctgctgcata tatactatca gaacagcatc gacgaccggc tacttgttga gcggatcttt
     3961 gctcagtaca tggtccctca caatttggaa acgacggaac ggatgaagtg cttgtattac
     4021 ttgtacgcta cactggactt gaatgctgtg aaagcattga atgaaatgtg gaaatgtcaa
     4081 aatctactcc gacatcaagt aaaggattta cttgatttaa ttaaacaacc aaaaacagat
     4141 gccagtgtca aggccatatt ttctaaagtg atggttatca caagaaattt gccagatcct
     4201 ggtaaggccc aagacttcat gaagaagttc acacaagtgt tggaagatga tgagaaaata
     4261 cggaaacagc tggaagccct tgttagtccc acatgttcct gcaagcaggc tgaaggctgt
     4321 gtgcgtgaaa taactaagaa gttgggcaac cctaagcagc ctacaaaccc tttcttggaa
     4381 atgatcaagt ttctcttgga gaggatagct cctgtacaca tagacactga atccatcagt
     4441 gctcttatca agcaagtgaa taaatcaata gatggaacag cagatgatga ggatgagggt
     4501 gttccaactg atcaagctat cagggcaggc cttgagctgc tgaaggtgct gtcttttacg
     4561 caccctatct cctttcactc tgctgagacg tttgagtccc ttctggcttg tctgaagatg
     4621 gatgatgaga aggtggcaga agctgcacta caaattttca aaaacacagg aagcaaaatt
     4681 gaagaggatt tccctcacat cagatcagcc ttgcttcctg tactacatca caaatccaaa
     4741 aaaggaccac ctcggcaggc caaatacgcc atccattgta ttcatgccat attttctagt
     4801 aaagaaaccc agtttgcaca gatatttgag cctctgcata aaagcctaga tccaagcaac
     4861 ctggagcatc ttataacccc tctggtcact attggtcata ttgctcttct cgcacctgat
     4921 cagttcgctg ctcctctgaa gtctttggtg gcaactttca ttgtgaagga cctcctcatg
     4981 aatgaccggc ttccaggaaa aaagacaact aagctttggg ttccagatga ggaagtctca
     5041 cctgagacaa tggtcaaaat tcaggctatt aaaatgatgg ttcgatggct acttggaatg
     5101 aagaataatc acagtaagtc aggaacttcc accctcagat tactaacaac gatactgcat
     5161 agtgatgggg atttgacaga acaagggaaa attagtaaac cagatatgtc acgcctgaga
     5221 cttgctgctg ggagtgctat tgtgaagctg gcacaggagc cctgttacca cgagatcatt
     5281 acactggagc agtaccagct gtgtgcatta gccatcaatg atgagtgtta tcaagtcagg
     5341 caggtgttcg ctcagaaact tcacaaaggc ctttcccgct tacggcttcc ccttgagtac
     5401 atggccatct gtgctctttg tgccaaagac cctgtgaaag agaggcgagc ccatgctaga
     5461 cagtgcttgg tgaagaacat cactgtgagg agggagtacc tgaagcagca tgcagctgtt
     5521 agtgaaaaat tattgtctct tctaccagag tatgttgttc catatacaat tcaccttttg
     5581 gcacatgacc cagattatgt caaagtacag gatattgaac aacttaaaga tgtgaaagaa
     5641 tgcctttggt ttgttctgga gatattgatg gctaaaaatg aaaacaacag ccatgcattt
     5701 atcagaaaaa tggtagaaaa tattaaacag acaaaagatg ctcaaggacc agatgataca
     5761 aaaatgaatg aaaaattgta caccgtgtgt gatgttgcca tgaacatcat catgtccaag
     5821 agcaccacgt acagcctgga gtctcctaag gaccccgtgc tgccagctcg gtttttcacc
     5881 cagcctgaca agaattttag taacaccaaa aattacctgc ctccagaaat gaaatcattt
     5941 ttcactcctg gaaaacctaa aacagccaat gttctcggag cagttaataa gccactttca
     6001 tcagcaggca aacagtctca gaccaaatca tcaagaatgg aaactgtgag caacgcaagc
     6061 agcagctcca acccaagctc tcctggaagg atcaagggga ggcttgatag ctctgaaatg
     6121 gatcacagtg aaaatgaaga ttatacaatg tcttcacctt tgccaggaaa aaaaagtgac
     6181 aagagagaag accctgatct ttctgagttg gagaagccta gaagtcggaa aaaagcacct
     6241 gtcacagacc ctgaagagaa attaggtatg gatgacctaa ctaagttggt acaggaacag
     6301 aaacctaaag gcagtcagcg aggccggaaa agaggccgta cagcttcaga ctcagacgag
     6361 cagcagtggc ctgaggagaa gaggcacaaa gaggagctcc tggaaaatga ggatgagcag
     6421 aacagcccac caaaaaaggg caaaagaggc aggccaccaa aacctcttgg tggggggaca
     6481 tcgaaggaag agcccacaat gaaaacatcc aagaaaggaa acaagaaaaa acttgtacct
     6541 cctgtagtag acgacgatga agaagaagaa agacaaattg gaaacacaga acataagtca
     6601 aaaagcaaac agcaccgcac atcaaagaga gcacaacaga gagcagaatc tcctgaaaca
     6661 agtgcagttg aatccacaca gtccacacca cagaaaggac gaggaagacc atcaaaagca
     6721 ccatcaccat cacaaccccc caaaaaaatc cgtgtaggac gctccaaaca ggtagccact
     6781 aaagaaaacg attcaagtga agaaatggat gtgcttcagg ccagctctcc tgtcagtgac
     6841 gacactacac aggaaggagc agaagaggag gacatttctg tgggaaatgt acgacggcga
     6901 agctccaaac gagagagacg atgagtttaa acgcggccgc tcaggtcagg tgcaggctgc
     6961 ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct
     7021 ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg
     7081 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     7141 cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag
     7201 tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca
     7261 tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg
     7321 aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt
     7381 ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc
     7441 tgtccctctt ctcttatgga gatccctcga cctgcagccc aagcttggcg taatcatggt
     7501 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     7561 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     7621 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgctgcatta
     7681 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     7741 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     7801 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     7861 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     7921 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7981 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     8041 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     8101 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     8161 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     8221 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     8281 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     8341 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     8401 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     8461 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     8521 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     8581 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     8641 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     8701 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     8761 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     8821 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     8881 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     8941 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     9001 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     9061 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     9121 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     9181 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     9241 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     9301 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     9361 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     9421 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     9481 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     9541 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     9601 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctg
//
