LOCUS       sequence-301984-        9652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For transient expression of human PDS5B tagged with mKate2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9652)
  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 9652)
  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: pEN765 -pCAGGS-3XFLAG-mKate2-(human)PDS5B-bpA
FEATURES             Location/Qualifiers
     source          1..9652
                     /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"
     primer_bind     complement(7013..7032)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7078..7133
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7132..7151)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7492..7508)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7492..7508)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7505..7527)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7516..7532
                     /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(7540..7546,7547..7564,7565..7570))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7585..7606
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7731..7748)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7902..8490)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7982..8001)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8661..9452,9453..9521))
                     /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     9284..9303
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9522..9626)
                     /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 actaggacca atgatggaaa aattacatat ccgcctgggg tcaaggaaat
     2641 atcagataaa atatctaaag aggagatggt gagacgatta aagatggttg tgaaaacttt
     2701 tatggatatg gaccaggact ctgaagaaga aaaggagctt tatttaaacc tagctttaca
     2761 tcttgcttca gatttttttc tcaagcatcc tgataaagat gttcgcttac tggtagcctg
     2821 ctgccttgct gatattttca ggatttatgc tcctgaagct ccttacacat cccctgataa
     2881 actaaaggat atatttatgt ttataacaag acagttgaag gggctagagg atacaaagag
     2941 cccacaattc aataggtatt tttatttact tgagaacatt gcttgggtca agtcatataa
     3001 catatgcttt gagttagaag atagcaatga aattttcacc cagctataca gaaccttatt
     3061 ttcagttata aacaatggcc acaatcagaa agtccatatg cacatggtag accttatgag
     3121 ctctattatt tgtgaaggtg atacagtgtc tcaggagctt ttggatacgg ttttagtaaa
     3181 tctggtacct gctcataaga atttaaacaa gcaagcatat gatttggcaa aggctttact
     3241 gaagaggaca gctcaagcta ttgagccata tattaccaat ttttttaatc aggttctgat
     3301 gcttgggaaa acatctatca gcgatttgtc agagcatgtc tttgacttaa ttttggagct
     3361 ctacaatatt gatagtcatt tgctgctctc tgttttaccc cagcttgaat ttaaattaaa
     3421 gagcaatgat aatgaggagc gcctacaagt tgttaaacta ctggcaaaaa tgtttggggc
     3481 aaaggattca gaattggctt ctcaaaacaa gccactttgg cagtgctact tgggcaggtt
     3541 taatgatatc catgtaccaa tccgcctgga atgtgtgaaa tttgctagcc attgtctcat
     3601 gaaccatcct gatttagcaa aagacttaac agagtatctt aaagtgaggt cacatgaccc
     3661 tgaggaagct attagacatg atgttattgt gtcaatagtt acagctgcta aaaaggatat
     3721 tcttctggtc aatgatcact tacttaattt tgtgagagag agaacattag acaaacgatg
     3781 gagagtacgc aaagaagcca tgatgggact tgcccaaatt tataagaaat atgctttaca
     3841 gtcagcagct ggaaaagatg ctgcaaaaca gatagcatgg atcaaagaca aattgctaca
     3901 tatatattat caaaatagta ttgatgatcg actacttgtt gaacggatct ttgctcaata
     3961 catggttcct cacaatttag aaactacaga acggatgaaa tgcttatatt acttgtatgc
     4021 cacactggat ttaaatgctg tgaaagcatt gaatgaaatg tggaaatgtc aaaatctgct
     4081 ccgacatcaa gtaaaggatt tgcttgactt gattaagcaa cccaaaacag atgccagtgt
     4141 caaggccata ttttcaaaag tgatggttat tacaagaaat ttacctgatc ctggtaaggc
     4201 tcaggatttc atgaagaaat tcacacaggt gttagaagat gatgagaaaa taagaaagca
     4261 gttagaagta cttgttagtc caacatgctc ctgcaagcag gctgaaggtt gtgtgcgtga
     4321 aataactaag aagttgggca accccaaaca gcctacaaat cctttcctgg aaatgatcaa
     4381 gtttctcttg gagaggatag cacctgtgca catagatacc gaatctatca gtgctcttat
     4441 taaacaagtg aacaaatcaa tagatggaac agcagatgat gaagatgagg gtgttccaac
     4501 tgatcaagcc atcagagcag gtcttgaact gcttaaggta ctctcattta cacatcccat
     4561 ctcatttcat tctgctgaaa catttgaatc attactggct tgtctgaaaa tggatgatga
     4621 aaaagtagca gaagctgcac tacaaatttt caaaaacaca ggaagcaaaa ttgaagagga
     4681 ttttccacac atcagatcag ccttgcttcc tgttttacat cacaaatcta aaaaaggacc
     4741 cccccgtcaa gccaaatatg ccattcattg tatccatgcg atattttcta gtaaagagac
     4801 ccagtttgca cagatatttg agcctctgca taagagccta gatccaagca acctggaaca
     4861 tctcataaca ccattggtta ctattggtca tattgctctc cttgcacctg atcaatttgc
     4921 tgctcctttg aaatctttgg tagctacttt cattgtgaaa gatcttctca tgaatgatcg
     4981 gcttccaggg aaaaagacaa ctaaactttg ggttccagat gaagaagtat ctcctgagac
     5041 aatggtcaaa attcaggcta ttaaaatgat ggttcgatgg ctacttggaa tgaaaaataa
     5101 tcacagtaaa tcaggaactt ctaccttaag attgctaaca acaatattgc atagtgatgg
     5161 agacttgaca gaacagggga aaattagtaa accagatatg tcacgtctga gacttgctgc
     5221 tgggagtgct attgtgaagc tggcacaaga accctgttac catgaaatca tcacattaga
     5281 acaatatcag ctatgtgcat tagctatcaa cgatgaatgc tatcaagtaa gacaagtgtt
     5341 tgcccagaaa cttcacaaag gcctttcccg tttacggctt ccacttgagt atatggcaat
     5401 ctgtgccctt tgtgcaaaag atcctgtaaa ggagagaaga gctcatgcta ggcaatgttt
     5461 ggtgaaaaat ataaatgtaa ggcgggagta tctgaagcag catgcagctg ttagtgaaaa
     5521 attattgtct cttctaccag agtatgttgt tccatataca attcaccttt tggcacatga
     5581 cccagattat gtcaaagtac aggatattga acaacttaaa gatgttaaag aatgtctttg
     5641 gtttgttctg gaaatattaa tggctaaaaa tgaaaataac agtcacgctt ttatcagaaa
     5701 gatggtagaa aatattaaac aaacaaaaga tgcccaagga ccagatgatg caaaaatgaa
     5761 tgaaaaactg tacactgtgt gtgatgttgc catgaatatc atcatgtcaa agagtactac
     5821 atacagtttg gaatctccta aagacccggt actaccagct cgtttcttca ctcaacctga
     5881 caagaatttc agtaacacca aaaattatct gcctcctgaa atgaaatcat ttttcactcc
     5941 tggaaaacct aaaacaacca atgttctagg agctgttaac aagccacttt catcagcagg
     6001 caagcaatct cagaccaaat catcacgaat ggaaactgta agcaatgcaa gcagcagctc
     6061 aaatccaagc tctcctggaa gaataaaggg gaggcttgat agttctgaaa tggatcacag
     6121 tgaaaatgaa gattacacaa tgtcttcacc tttgccgggg aaaaaaagtg acaagagaga
     6181 cgactctgat cttgtaaggt ctgaattgga gaagcctaga ggcaggaaaa aaacgcccgt
     6241 cacagaacag gaggagaaat taggtatgga tgacttgact aagttggtac aggaacagaa
     6301 acctaaaggc agtcagcgaa gtcggaaaag aggccatacg gcttcagaat ctgatgaaca
     6361 gcagtggcct gaggaaaaga ggctcaaaga agatatatta gaaaatgaag atgaacagaa
     6421 tagtccgcca aaaaagggta aaagaggccg accaccaaaa cctcttggtg gaggtacacc
     6481 aaaagaagag ccaacaatga aaacttctaa aaaaggaagc aaaaaaaaat ctggacctcc
     6541 agcaccagag gaggaggaag aagaagaaag acaaagtgga aatacggaac agaagtccaa
     6601 aagcaaacag caccgagtgt caaggagagc acagcagaga gcagaatctc ctgaatctag
     6661 tgcaattgaa tccacacagt ccacaccaca gaaaggacga ggaagaccat caaaaacgcc
     6721 atcaccatca caaccaaaaa aaaatgtccg tgtaggacgc tccaaacaag cagctactaa
     6781 ggaaaatgat tcaagtgaag aagtagatgt gtttcagggt agctctcctg tcgatgatat
     6841 tccacaggaa gaaacagagg aggaggaagt ttctacagta aatgtacggc ggcgaagtgc
     6901 taaaagggaa cggcgatgag tttaaacgcg gccgctcagg tcaggtgcag gctgcctatc
     6961 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     7021 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     7081 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     7141 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     7201 caacatatgc catatgctgg ctgccatgaa caaaggtggc tataaagagg tcatcagtat
     7261 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
     7321 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
     7381 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
     7441 cttctcttat gaagatccct cgacctgcag cccaagcttg gcgtaatcat ggtcatagct
     7501 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     7561 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     7621 actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgctgca ttaatgaatc
     7681 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     7741 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     7801 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     7861 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     7921 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     7981 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     8041 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     8101 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     8161 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     8221 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     8281 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     8341 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     8401 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     8461 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     8521 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     8581 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     8641 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     8701 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     8761 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     8821 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     8881 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     8941 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     9001 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     9061 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     9121 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     9181 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     9241 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     9301 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     9361 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     9421 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     9481 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     9541 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     9601 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tg
//
