LOCUS       sequence-301988-        9307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For transient expression of mouse PDS5A tagged with mKate2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9307)
  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 9307)
  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: pKS016 - pCAGGS-3XFLAG-mKate2-Pds5a-bpA
FEATURES             Location/Qualifiers
     source          1..9307
                     /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(6668..6687)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6733..6788
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6787..6806)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7147..7163)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7147..7163)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7160..7182)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7171..7187
                     /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(7195..7201,7202..7219,7220..7225))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7240..7261
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7386..7403)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7557..8145)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7637..7656)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8316..9107,9108..9176))
                     /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     8939..8958
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9177..9281)
                     /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 cttcacgcag ccgaagcctg ccactgccct ctgtggcgtc gtgagtgcgg acgggaagat
     2641 cgcttaccct ccgggagtaa aagagatcac cgacaagatc accaccgatg aaatgatcaa
     2701 acgtctgaag atggtagtga aaacttttat ggatatggat caggattcag aagatgaaaa
     2761 gcaacagtat ctcccactag ccttgcatct cgcatctgaa ttctttctca ggaaccctaa
     2821 taaagatgtg cgccttctcg tagcatgttg tttggctgat atatttcgaa tctatgcccc
     2881 agaagctcca tatacttccc atgataaact taaggacata tttctcttta ttaccagaca
     2941 gttaaaaggt ttggaggata caaagagtcc acagtttaat agatattttt atttattaga
     3001 gaatttagct tgggttaaat catataacat ctgctttgag ttggaagatt gcaatgaaat
     3061 ttttattcag ctttttagaa ctctcttctc ggtaatcaac aatagccaca atacgaaagt
     3121 acaaatgcac atgttagact tgatgagttc tatcatcatg gaaggtgatg gagttactca
     3181 agaattattg gattccattc ttatcaacct catccctgca cacaagaact taaataaaca
     3241 gtccttcgac cttgcaaaag tcctactgaa aaggacagtc cagactattg aagcatgtat
     3301 tgccaatttt ttcaatcaag tcctggtgct gggcagatca tcagtcagcg acctgtctga
     3361 acacgtattt gatctgattc aggaactttt tgctatcgat cctcagttac tgttatctgt
     3421 catgccacag cttgaattca aactgaagag caacgatggt gaagaacgcc tagctgtggt
     3481 tcgactcctc gcaaaattgt tcggctctaa agattcagat ttagcaacac agaatcggcc
     3541 tctctggcag tgctttcttg ggcgatttaa tgacattcat gttcctgtga ggttagaaag
     3601 tgtgaagttt gccagccact gtttgatgaa tcacccagac ttagcgaagg atctgacaga
     3661 atatttgaaa gttaggtcac atgatccaga agaagccatt cgtcatgatg ttattgttac
     3721 tataataaca gctgccaaaa gagaccttgc cttagtaaat gatcagttgc ttggctttgt
     3781 cagggaaagg acactggata aacggtggcg agtaagaaaa gaagccatga tgggtctggc
     3841 tcagctctat aagaaatact gtcttcatgg ggaagcagga aaggaagccg cggagaaagt
     3901 cagctggata aaggacaagc tgctgcatat ctactatcag aacagcattg atgacaaact
     3961 attggtagag aaaatctttg ctcagtatct tgttccccac aacctggaaa cagaagagag
     4021 aatgaaatgc ttatattatt tatatgctag cttggatcca aatgctgtaa aagctctcaa
     4081 tgaaatgtgg aaatgtcaga acatgcttcg aagtcatgtg cgtgaactat tggacttaca
     4141 caagcagcct acatcagaag cgaactgttc tgccatgttt gggaaactga tgaccatagc
     4201 aaagaatttg cctgaccctg gaaaagcaca agattttgta aagaaattta accaggttct
     4261 tggtgatgat gagaaactga ggtctcagtt ggaattatta atcagcccaa cctgttcatg
     4321 caagcaggct gacgtttgtg tgagggaaat agctcggaaa cttgcaaatc ctaagcagcc
     4381 aaccaaccct tttctagaga tggtcaaatt tcttttggaa agaattgctc ctgtgcacat
     4441 cgattcagaa gccataagtg cactggtaaa actgatgaat aagtcaatcg aagggacggc
     4501 cgatgatgaa gaggagggtg tcagtccaga ctcagccatc cgctcaggac ttgagcttct
     4561 taaggttctg tctttcacac atcctacctc gttccactct gcagagacat atgagtcctt
     4621 gttacagtgc ctaagaatgg aggatgacaa ggtagcagaa gctgcaatac aaatttttag
     4681 aaacacaggc cacaaaatag aaactgacct tccccagata cggtcaacct tgatccccat
     4741 tttacatcag aaagccaagc ggggcactcc acaccaagca aagcaggctg ttcactgcat
     4801 ccatgccatc ttctcaaaca aggaggtcca gctggcacag atttttgagc cactcagtag
     4861 gagtctgaat gctgatgtac cagagcaact tataactcca ttagtttcac tgggccacat
     4921 ttccatgtta gcaccagatc agtttgcctc cccgatgaaa tctgtagtgg caaactttat
     4981 tgttaaagac cttctaatga acgacaggtc aacaggtgag aagaatggaa aattatggtc
     5041 tccagatgag gaagtttctc ctgaagttct agcaaaggta caggcaatta aacttctggt
     5101 aaggtggctg ttgggtatga aaaacaacca gtctaaatct gccaactcaa ctcttcggtt
     5161 attatcagcc atgctggtta gtgagggcga cctgacagag caaaagagga tcagtaaatc
     5221 tgatatgtct cgcttgcgat tagctgctgg tagtgccata atgaagcttg ctcaggaacc
     5281 ttgctaccat gaaattatta ccccagaaca gtttcagctc tgtgcactgg ttattaatga
     5341 tgagtgctac caagtaaggc agatatttgc ccagaagctt cataaggcac ttgtgaagtt
     5401 gcttctccca ctggagtata tggccatctt tgctttgtgt gccaaagacc ctgtgaaaga
     5461 aaggagagca cacgctcggc agtgtttgtt aaagaacatc agcatccgca gggagtacat
     5521 caaacagaac cccatggcca ctgagaaatt attgtcactg ctgcctgaat atgtggttcc
     5581 atacatgatt cacctcctag cccatgatcc tgatttcaca cggtcacaag atgttgatca
     5641 acttcgtgat ataaaagagt gcctgtggtt tatgcttgaa gtcttaatga caaagaatga
     5701 aaacaacagc catgcattca tgaaaaagat ggcagagaat atcaagctaa ccagagacgc
     5761 ccagtctccc gatgaatcca agacaaatga aaaactttat acggtttgtg acgtggctct
     5821 gtgtgttata aatagtaaaa gtgctttgtg caatgcagac tcaccaaagg acccagtcct
     5881 cccaatgaag ttctttacac agcctgaaaa ggacttctgt aatgacaaaa gctatatttc
     5941 agaagagaca agagttcttc tgttaacagg aaagccaaag cctactggag tattaggtac
     6001 agtgaacaag cctttatcag caacgggaag gaagccttat gttagaagcg ccggcacaga
     6061 gactggaagc aatattaacg ccaattcaga gctgagtcct tcagccggaa gtcgttcaag
     6121 ggaacagagt tcagaggcat cagaaactgg agttagtgaa aatgaggaga atcctgtgag
     6181 aataatttct gtcacacctg taaagaatat tgatactgta aagaataagg aaattaattc
     6241 tgatcagtct acccaaggca acatcagcag tgacagagga aagaaaagaa ttgtaacagc
     6301 agctggtgca gagaatatcc aaaagccaga cgagaaagta gatgagtcag gaccgcctgc
     6361 cccttccaaa ccccggagag gacgtcgccc caaatctgaa tctcagggca atgcaaccaa
     6421 aaacgatgat ctaaataaac ctgttagcaa gggaaggaaa agagctgcag gcagccagga
     6481 gagtctggag gcagggaatg ccaaagcgcc caagctacaa gatggagcca aaaaggcagt
     6541 tccagcagag agacaaattg atttacaaag gtaagtttaa acgcggccgc tcaggtcagg
     6601 tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc
     6661 actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc
     6721 tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
     6781 gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt
     6841 tggtttagag tttggcaaca tatgccatat gctggctgcc atgaacaaag gtggctataa
     6901 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     6961 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     7021 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     7081 gtcatagctg tccctcttct cttatgaaga tccctcgacc tgcagcccaa gcttggcgta
     7141 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     7201 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     7261 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg
     7321 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     7381 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     7441 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     7501 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7561 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7621 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     7681 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     7741 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     7801 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7861 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7921 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7981 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     8041 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     8101 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     8161 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     8221 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     8281 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     8341 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     8401 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     8461 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     8521 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     8581 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     8641 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     8701 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     8761 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     8821 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     8881 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     8941 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     9001 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     9061 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     9121 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     9181 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     9241 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     9301 ccacctg
//
