LOCUS       sequence-301997-        9322 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For transient expression of human PDS5A tagged with mKate2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9322)
  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 9322)
  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: pKS071 - pCAGGS-3XFLAG-mKate2-(human)PDS5A-bpA
FEATURES             Location/Qualifiers
     source          1..9322
                     /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(6683..6702)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6748..6803
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6802..6821)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7162..7178)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7162..7178)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7175..7197)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7186..7202
                     /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(7210..7216,7217..7234,7235..7240))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7255..7276
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7401..7418)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7572..8160)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7652..7671)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8331..9122,9123..9191))
                     /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     8954..8973
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9192..9296)
                     /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 ccggtatgga
     2581 cttcaccgcg cagcccaagc ctgccactgc cctctgtggc gtcgtgagtg ccgacgggaa
     2641 gatcgcttac cctccggggg taaaagagat caccgacaag atcaccacgg acgagatgat
     2701 caaacgcctg aagatggtag tgaaaacctt tatggatatg gatcaggact cagaagatga
     2761 aaaacagcag tatctcccac tagccttgca tcttgcatct gaattcttcc tcaggaaccc
     2821 caataaagat gtgcgtctcc ttgtagcatg ttgtttggct gatatctttc gtatctatgc
     2881 cccagaagct ccatatactt cccatgataa acttaaggac atatttttgt ttattaccag
     2941 acaattaaaa ggtttggagg atacaaagag tccacagttt aatagatact tttatttatt
     3001 agagaattta gcttgggtta aatcatataa catctgcttt gaattggaag attgcaatga
     3061 aatttttatt cagcttttta gaactctctt ctcagtgatc aacaatagcc acaataagaa
     3121 ggtacaaatg cacatgctag atttgatgag ttctatcatc atggaaggtg atggagttac
     3181 tcaagaatta ttggactcca ttcttattaa cctcattcct gcacataaga acttaaataa
     3241 acagtccttt gaccttgcaa aagtgctatt gaaaagaaca gtccagacta ttgaggcatg
     3301 cattgctaat tttttcaatc aagtcctggt gctgggaaga tcatcagtaa gtgatttgtc
     3361 agaacatgta tttgatctga ttcaggaact ttttgctata gatcctcatt tattattatc
     3421 cgtcatgcca cagcttgaat tcaaactaaa gagcaatgat ggagaagagc gattagctgt
     3481 tgttcgactt ctagctaaat tgtttggctc caaagattct gatttggcaa cacagaatcg
     3541 tcctctttgg caatgttttc ttggacgatt taatgatatt catgttcctg tgagattaga
     3601 aagtgtgaaa tttgccagtc attgtttaat gaatcaccca gatttagcga aggatctcac
     3661 agaatattta aaggttagat cacatgatcc agaagaagct attcgtcatg atgtcattgt
     3721 tactataata acagctgcca agagggacct ggccttagta aatgatcagc tgcttggctt
     3781 tgtaagggaa agaacactgg ataaacggtg gcgagtaaga aaagaagcta tgatgggtct
     3841 ggctcagctt tataagaaat actgtcttca tggtgaagca ggaaaggaag ctgcagagaa
     3901 agtcagctgg ataaaggaca aacttctgca tatttattat cagaacagca ttgacgacaa
     3961 actgttggta gagaaaatct ttgctcagta tcttgtcccc cacaacctgg aaacagaaga
     4021 gagaatgaaa tgcttatatt acttatatgc tagtttggat ccaaatgctg taaaagctct
     4081 caacgaaatg tggaagtgtc agaacatgct tcggagccat gtacgcgaac tattggattt
     4141 gcacaagcag cctacatcag aggctaactg ttctgccatg tttggaaaac tgatgaccat
     4201 agcaaagaat ttgcctgacc ccgggaaagc acaagatttt gtgaagaaat ttaaccaggt
     4261 tctcggcgat gatgagaaac ttcggtctca gttggagtta ttaattagcc caacctgttc
     4321 ttgcaaacaa gcagatattt gtgtgagaga aatagcccgg aaacttgcaa atcctaagca
     4381 accaacaaat ccttttctag agatggtcaa atttctgttg gaaagaatcg cacctgtgca
     4441 cattgattca gaagccataa gtgcactagt gaaattgatg aataagtcaa tagaggggac
     4501 agcagatgat gaagaggagg gtgtaagtcc agatacagct atccgttcag gacttgaact
     4561 tcttaaggtt ctgtctttta cacatcctac ctcgttccac tctgcagaga catatgagtc
     4621 cttgttacag tgcctaagaa tggaggatga caaggtagca gaagctgcta ttcaaatttt
     4681 tagaaataca ggtcacaaaa tagaaacaga ccttccccag atacgatcga ccttaattcc
     4741 cattttacat caaaaagcaa agaggggtac tccacaccaa gcaaaacagg ctgtgcactg
     4801 tatacacgcc atattcacaa ataaagaagt ccagcttgca cagatttttg agccactcag
     4861 taggagtctg aatgctgatg tgccagaaca acttataact ccattagttt cattgggcca
     4921 catttctatg ttagcaccag atcagtttgc ttccccaatg aaatctgtag tagcaaattt
     4981 tattgtgaaa gatctgctaa tgaatgacag gtcaacaggt gaaaagaatg gaaaactgtg
     5041 gtctccagat gaagaggttt cccctgaagt actagcaaag gtacaggcaa ttaaacttct
     5101 ggtaaggtgg ctgttgggta tgaaaaacaa ccagtctaaa tctgccaatt caacccttcg
     5161 gttattatca gcgatgttgg ttagtgaggg tgacctgaca gagcaaaaga ggatcagtaa
     5221 atctgatatg tctcgcttgc gattagctgc tggtagtgcc ataatgaagc ttgctcagga
     5281 accttgttac catgaaatta ttaccccaga acagtttcag ctctgtgcac ttgttattaa
     5341 tgatgagtgt taccaagtaa ggcagatatt tgctcagaag ctgcataagg cacttgtgaa
     5401 gttactgctc ccattggagt atatggcgat ctttgccttg tgtgccaaag atcctgtgaa
     5461 ggagagaaga gcacacgcac gacaatgttt actgaaaaat atcagtatac gcagggaata
     5521 cattaagcag aatcctatgg ctactgagaa attattatca ctgttgcctg aatatgtagt
     5581 tccatacatg attcacctgc tagcccatga tccagatttt acaagatcac aagatgttga
     5641 tcagcttcgt gatatcaaag agtgcctatg gttcatgctt gaagttttaa tgacaaagaa
     5701 tgaaaacaat agccatgcct ttatgaagaa gatggcagag aacatcaagt taaccagaga
     5761 tgcccagtct ccagatgaat ccaagacaaa tgaaaaactg tatacagtat gtgatgtggc
     5821 tctctgtgtt ataaatagta aaagtgcttt gtgcaatgca gattcaccaa aggacccagt
     5881 cctcccaatg aaatttttta cacaacctga aaaggacttc tgtaacgata agagttatat
     5941 ttcagaagag acaagagtac ttctgttaac aggaaagcca aagcctgctg gagtactagg
     6001 tgcagtaaat aagcctttat cagcaacggg aaggaaaccc tatgttagaa gcactggcac
     6061 tgagactgga agcaatatta atgtaaattc agagctgaac ccttcaaccg gaaatcgatc
     6121 aagggaacag agttcagagg cagcagaaac tggagttagt gaaaatgaag agaaccctgt
     6181 gaggattatt tcagtcacac ctgtaaagaa tattgaccca gtaaagaata aggaaattaa
     6241 ttctgatcag gctacccagg gcaacatcag cagtgaccga ggaaagaaaa gaacagtaac
     6301 agcagctggt gcagagaata tccaacaaaa aacagatgag aaagtagatg aatcgggacc
     6361 tcccgcccct tccaaaccca ggagaggacg tcgacccaag tctgaatctc agggcaatgc
     6421 taccaaaaat gatgatctaa ataaacctat taacaaggga aggaagagag ctgcagtggg
     6481 tcaggagagc cctgggggtt tggaagcagg taatgccaaa gcacccaaac tgcaagattt
     6541 agccaaaaag gcagcaccag cagaaagaca aattgactta caaaggtaag tttaaacgcg
     6601 gccgctcagg tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc
     6661 tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga
     6721 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
     6781 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
     6841 atcagaatga gtatttggtt tagagtttgg caacatatgc catatgctgg ctgccatgaa
     6901 caaaggtggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
     6961 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
     7021 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
     7081 tcctgactac tcccagtcat agctgtccct cttctcttat gaagatccct cgacctgcag
     7141 cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     7201 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     7261 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     7321 gtgccagcgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     7381 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     7441 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     7501 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     7561 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     7621 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     7681 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7741 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     7801 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7861 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7921 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7981 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     8041 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     8101 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     8161 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     8221 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     8281 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     8341 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     8401 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     8461 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     8521 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     8581 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     8641 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     8701 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8761 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8821 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8881 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8941 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     9001 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     9061 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     9121 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     9181 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     9241 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     9301 tttccccgaa aagtgccacc tg
//
