LOCUS       sequence-310154-        9662 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 9662)
  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 9662)
  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..9662
                     /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"
     primer_bind     complement(7021..7040)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7086..7141
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7140..7159)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7502..7518)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7502..7518)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7515..7537)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7526..7542
                     /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(7550..7556,7557..7574,7575..7580))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7595..7616
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7741..7758)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7912..8500)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7992..8011)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8671..9462,9463..9531))
                     /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     9294..9313
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9532..9636)
                     /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 gsggggsggg 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 taggaccaat gatggaaaaa ttacatatcc gcctggggtc
     2641 aaggaaatat cagataaaat atctaaagag gagatggtga gacgattaaa gatggttgtg
     2701 aaaactttta tggatatgga ccaggactct gaagaagaaa aggagcttta tttaaaccta
     2761 gctttacatc ttgcttcaga tttttttctc aagcatcctg ataaagatgt tcgcttactg
     2821 gtagcctgct gccttgctga tattttcagg atttatgctc ctgaagctcc ttacacatcc
     2881 cctgataaac taaaggatat atttatgttt ataacaagac agttgaaggg gctagaggat
     2941 acaaagagcc cacaattcaa taggtatttt tatttacttg agaacattgc ttgggtcaag
     3001 tcatataaca tatgctttga gttagaagat agcaatgaaa ttttcaccca gctatacaga
     3061 accttatttt cagttataaa caatggccac aatcagaaag tccatatgca catggtagac
     3121 cttatgagct ctattatttg tgaaggtgat acagtgtctc aggagctttt ggatacggtt
     3181 ttagtaaatc tggtacctgc tcataagaat ttaaacaagc aagcatatga tttggcaaag
     3241 gctttactga agaggacagc tcaagctatt gagccatata ttaccaattt ttttaatcag
     3301 gttctgatgc ttgggaaaac atctatcagc gatttgtcag agcatgtctt tgacttaatt
     3361 ttggagctct acaatattga tagtcatttg ctgctctctg ttttacccca gcttgaattt
     3421 aaattaaaga gcaatgataa tgaggagcgc ctacaagttg ttaaactact ggcaaaaatg
     3481 tttggggcaa aggattcaga attggcttct caaaacaagc cactttggca gtgctacttg
     3541 ggcaggttta atgatatcca tgtaccaatc cgcctggaat gtgtgaaatt tgctagccat
     3601 tgtctcatga accatcctga tttagcaaaa gacttaacag agtatcttaa agtgaggtca
     3661 catgaccctg aggaagctat tagacatgat gttattgtgt caatagttac agctgctaaa
     3721 aaggatattc ttctggtcaa tgatcactta cttaattttg tgagagagag aacattagac
     3781 aaacgatgga gagtacgcaa agaagccatg atgggacttg cccaaattta taagaaatat
     3841 gctttacagt cagcagctgg aaaagatgct gcaaaacaga tagcatggat caaagacaaa
     3901 ttgctacata tatattatca aaatagtatt gatgatcgac tacttgttga acggatcttt
     3961 gctcaataca tggttcctca caatttagaa actacagaac ggatgaaatg cttatattac
     4021 ttgtatgcca cactggattt aaatgctgtg aaagcattga atgaaatgtg gaaatgtcaa
     4081 aatctgctcc gacatcaagt aaaggatttg cttgacttga ttaagcaacc caaaacagat
     4141 gccagtgtca aggccatatt ttcaaaagtg atggttatta caagaaattt acctgatcct
     4201 ggtaaggctc aggatttcat gaagaaattc acacaggtgt tagaagatga tgagaaaata
     4261 agaaagcagt tagaagtact tgttagtcca acatgctcct gcaagcaggc tgaaggttgt
     4321 gtgcgtgaaa taactaagaa gttgggcaac cccaaacagc ctacaaatcc tttcctggaa
     4381 atgatcaagt ttctcttgga gaggatagca cctgtgcaca tagataccga atctatcagt
     4441 gctcttatta aacaagtgaa caaatcaata gatggaacag cagatgatga agatgagggt
     4501 gttccaactg atcaagccat cagagcaggt cttgaactgc ttaaggtact ctcatttaca
     4561 catcccatct catttcattc tgctgaaaca tttgaatcat tactggcttg tctgaaaatg
     4621 gatgatgaaa aagtagcaga agctgcacta caaattttca aaaacacagg aagcaaaatt
     4681 gaagaggatt ttccacacat cagatcagcc ttgcttcctg ttttacatca caaatctaaa
     4741 aaaggacccc cccgtcaagc caaatatgcc attcattgta tccatgcgat attttctagt
     4801 aaagagaccc agtttgcaca gatatttgag cctctgcata agagcctaga tccaagcaac
     4861 ctggaacatc tcataacacc attggttact attggtcata ttgctctcct tgcacctgat
     4921 caatttgctg ctcctttgaa atctttggta gctactttca ttgtgaaaga tcttctcatg
     4981 aatgatcggc ttccagggaa aaagacaact aaactttggg ttccagatga agaagtatct
     5041 cctgagacaa tggtcaaaat tcaggctatt aaaatgatgg ttcgatggct acttggaatg
     5101 aaaaataatc acagtaaatc aggaacttct accttaagat tgctaacaac aatattgcat
     5161 agtgatggag acttgacaga acaggggaaa attagtaaac cagatatgtc acgtctgaga
     5221 cttgctgctg ggagtgctat tgtgaagctg gcacaagaac cctgttacca tgaaatcatc
     5281 acattagaac aatatcagct atgtgcatta gctatcaacg atgaatgcta tcaagtaaga
     5341 caagtgtttg cccagaaact tcacaaaggc ctttcccgtt tacggcttcc acttgagtat
     5401 atggcaatct gtgccctttg tgcaaaagat cctgtaaagg agagaagagc tcatgctagg
     5461 caatgtttgg tgaaaaatat aaatgtaagg cgggagtatc tgaagcagca tgcagctgtt
     5521 agtgaaaaat tattgtctct tctaccagag tatgttgttc catatacaat tcaccttttg
     5581 gcacatgacc cagattatgt caaagtacag gatattgaac aacttaaaga tgttaaagaa
     5641 tgtctttggt ttgttctgga aatattaatg gctaaaaatg aaaataacag tcacgctttt
     5701 atcagaaaga tggtagaaaa tattaaacaa acaaaagatg cccaaggacc agatgatgca
     5761 aaaatgaatg aaaaactgta cactgtgtgt gatgttgcca tgaatatcat catgtcaaag
     5821 agtactacat acagtttgga atctcctaaa gacccggtac taccagctcg tttcttcact
     5881 caacctgaca agaatttcag taacaccaaa aattatctgc ctcctgaaat gaaatcattt
     5941 ttcactcctg gaaaacctaa aacaaccaat gttctaggag ctgttaacaa gccactttca
     6001 tcagcaggca agcaatctca gaccaaatca tcacgaatgg aaactgtaag caatgcaagc
     6061 agcagctcaa atccaagctc tcctggaaga ataaagggga ggcttgatag ttctgaaatg
     6121 gatcacagtg aaaatgaaga ttacacaatg tcttcacctt tgccggggaa aaaaagtgac
     6181 aagagagacg actctgatct tgtaaggtct gaattggaga agcctagagg caggaaaaaa
     6241 acgcccgtca cagaacagga ggagaaatta ggtatggatg acttgactaa gttggtacag
     6301 gaacagaaac ctaaaggcag tcagcgaagt cggaaaagag gccatacggc ttcagaatct
     6361 gatgaacagc agtggcctga ggaaaagagg ctcaaagaag atatattaga aaatgaagat
     6421 gaacagaata gtccgccaaa aaagggtaaa agaggccgac caccaaaacc tcttggtgga
     6481 ggtacaccaa aagaagagcc aacaatgaaa acttctaaaa aaggaagcaa aaaaaaatct
     6541 ggacctccag caccagagga ggaggaagaa gaagaaagac aaagtggaaa tacggaacag
     6601 aagtccaaaa gcaaacagca ccgagtgtca aggagagcac agcagagagc agaatctcct
     6661 gaatctagtg caattgaatc cacacagtcc acaccacaga aaggacgagg aagaccatca
     6721 aaaacgccat caccatcaca accaaaaaaa aatgtccgtg taggacgctc caaacaagca
     6781 gctactaagg aaaatgattc aagtgaagaa gtagatgtgt ttcagggtag ctctcctgtc
     6841 gatgatattc cacaggaaga aacagaggag gaggaagttt ctacagtaaa tgtacggcgg
     6901 cgaagtgcta aaagggaacg gcgatgagtt taaacgcggc cgctcaggtc aggtgcaggc
     6961 tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga
     7021 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
     7081 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     7141 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
     7201 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg
     7261 tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac
     7321 ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa
     7381 attttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat
     7441 agctgtccct cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat
     7501 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     7561 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     7621 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgctgca
     7681 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     7741 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     7801 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     7861 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     7921 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7981 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     8041 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     8101 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     8161 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     8221 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     8281 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     8341 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     8401 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     8461 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     8521 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     8581 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     8641 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     8701 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     8761 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     8821 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8881 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8941 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     9001 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     9061 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     9121 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     9181 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     9241 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     9301 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     9361 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     9421 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     9481 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     9541 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     9601 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     9661 tg
//
