LOCUS       sequence-310133-        9331 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 9331)
  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 9331)
  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..9331
                     /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(6690..6709)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6755..6810
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6809..6828)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7171..7187)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7171..7187)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7184..7206)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7195..7211
                     /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(7219..7225,7226..7243,7244..7249))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7264..7285
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7410..7427)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7581..8169)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7661..7680)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8340..9131,9132..9200))
                     /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     8963..8982
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9201..9305)
                     /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 ggtatggact tcaccgcgca gcccaagcct gccactgccc tctgtggcgt cgtgagtgcc
     2641 gacgggaaga tcgcttaccc tccgggggta aaagagatca ccgacaagat caccacggac
     2701 gagatgatca aacgcctgaa gatggtagtg aaaaccttta tggatatgga tcaggactca
     2761 gaagatgaaa aacagcagta tctcccacta gccttgcatc ttgcatctga attcttcctc
     2821 aggaacccca ataaagatgt gcgtctcctt gtagcatgtt gtttggctga tatctttcgt
     2881 atctatgccc cagaagctcc atatacttcc catgataaac ttaaggacat atttttgttt
     2941 attaccagac aattaaaagg tttggaggat acaaagagtc cacagtttaa tagatacttt
     3001 tatttattag agaatttagc ttgggttaaa tcatataaca tctgctttga attggaagat
     3061 tgcaatgaaa tttttattca gctttttaga actctcttct cagtgatcaa caatagccac
     3121 aataagaagg tacaaatgca catgctagat ttgatgagtt ctatcatcat ggaaggtgat
     3181 ggagttactc aagaattatt ggactccatt cttattaacc tcattcctgc acataagaac
     3241 ttaaataaac agtcctttga ccttgcaaaa gtgctattga aaagaacagt ccagactatt
     3301 gaggcatgca ttgccaattt tttcaatcaa gtcctggtgc tgggaagatc atcagtaagt
     3361 gatttgtcag aacatgtatt tgatctgatt caggaacttt ttgctataga tcctcattta
     3421 ttattatccg tcatgccaca gcttgaattc aaactaaaga gcaatgatgg agaagagcga
     3481 ttagctgttg ttcgacttct agctaaattg tttggctcca aagattctga tttggcaaca
     3541 cagaatcgtc ctctttggca atgttttctt ggacgattta atgatattca tgttcctgtg
     3601 agattagaaa gtgtgaaatt tgccagtcat tgtttaatga atcacccaga tttagcgaag
     3661 gatctcacag aatatttaaa ggttagatca catgatccag aagaagctat tcgtcatgat
     3721 gtcattgtta ctataataac agctgccaag agggacctgg ccttagtaaa tgatcagctg
     3781 cttggctttg taagggaaag aacactggat aaacggtggc gagtaagaaa agaagctatg
     3841 atgggtctgg ctcagcttta taagaaatac tgtcttcatg gtgaagcagg aaaggaagct
     3901 gcagagaaag tcagctggat aaaggacaaa cttctgcata tttattatca gaacagcatt
     3961 gacgacaaac tgttggtaga gaaaatcttt gctcagtatc ttgtccccca caacctggaa
     4021 acagaagaga gaatgaaatg cttatattac ttatatgcta gtttggatcc aaatgctgta
     4081 aaagctctca acgaaatgtg gaagtgtcag aacatgcttc ggagccatgt acgcgaacta
     4141 ttggatttgc acaagcagcc tacatcagag gctaactgtt ctgccatgtt tggaaaactg
     4201 atgaccatag caaagaattt gcctgacccc gggaaagcac aagattttgt gaagaaattt
     4261 aaccaggttc tcggcgatga tgagaaactt cggtctcagt tggagttatt aattagccca
     4321 acctgttctt gcaaacaagc agatatttgt gtgagagaaa tagcccggaa acttgcaaat
     4381 cctaagcaac caacaaatcc ttttctagag atggtcaaat ttctgttgga aagaatcgca
     4441 cctgtgcaca ttgattcaga agccataagt gcactagtga aattgatgaa taagtcaata
     4501 gaggggacag cagatgatga agaggagggt gtaagtccag atacagctat ccgttcagga
     4561 cttgaacttc ttaaggttct gtcttttaca catcctacct cgttccactc tgcagagaca
     4621 tatgagtcct tgttacagtg cctaagaatg gaggatgaca aggtagcaga agctgctatt
     4681 caaattttta gaaatacagg tcacaaaata gaaacagacc ttccccagat acgatcgacc
     4741 ttaattccca ttttacatca aaaagcaaag aggggtactc cacaccaagc aaaacaggct
     4801 gtgcactgta tacacgccat attcacaaat aaagaagtcc agcttgcaca gatttttgag
     4861 ccactcagta ggagtctgaa tgctgatgtg ccagaacaac ttataactcc attagtttca
     4921 ttgggccaca tttctatgtt agcaccagat cagtttgctt ccccaatgaa atctgtagta
     4981 gcaaatttta ttgtgaaaga tctgctaatg aatgacaggt caacaggtga aaagaatgga
     5041 aaactgtggt ctccagatga agaggtttcc cctgaagtac tagcaaaggt acaggcaatt
     5101 aaacttctgg taaggtggct gttgggtatg aaaaacaacc agtctaaatc tgccaattca
     5161 acccttcggt tattatcagc gatgttggtt agtgagggtg acctgacaga gcaaaagagg
     5221 atcagtaaat ctgatatgtc tcgcttgcga ttagctgctg gtagtgccat aatgaagctt
     5281 gctcaggaac cttgttacca tgaaattatt accccagaac agtttcagct ctgtgcactt
     5341 gttattaatg atgagtgtta ccaagtaagg cagatatttg ctcagaagct gcataaggca
     5401 cttgtgaagt tactgctccc attggagtat atggcgatct ttgccttgtg tgccaaagat
     5461 cctgtgaagg agagaagagc acacgcacga caatgtttac tgaaaaatat cagtatacgc
     5521 agggaataca ttaagcagaa tcctatggct actgagaaat tattatcact gttgcctgaa
     5581 tatgtagttc catacatgat tcacctgcta gcccatgatc cagattttac aagatcacaa
     5641 gatgttgatc agcttcgtga tatcaaagag tgcctatggt tcatgcttga agttttaatg
     5701 acaaagaatg aaaacaatag ccatgccttt atgaagaaga tggcagagaa catcaagtta
     5761 accagagatg cccagtctcc agatgaatcc aagacaaatg aaaaactgta tacagtatgt
     5821 gatgtggctc tctgtgttat aaatagtaaa agtgctttgt gcaatgcaga ttcaccaaag
     5881 gacccagtcc tcccaatgaa attttttaca caacctgaaa aggacttctg taacgataag
     5941 agttatattt cagaagagac aagagtactt ctgttaacag gaaagccaaa gcctgctgga
     6001 gtactaggtg cagtaaataa gcctttatca gcaacgggaa ggaaacccta tgttagaagc
     6061 actggcactg agactggaag caatattaat gtaaattcag agctgaaccc ttcaaccgga
     6121 aatcgatcaa gggaacagag ttcagaggca gcagaaactg gagttagtga aaatgaagag
     6181 aaccctgtga ggattatttc agtcacacct gtaaagaata ttgacccagt aaagaataag
     6241 gaaattaatt ctgatcaggc tacccagggc aacatcagca gtgaccgagg aaagaaaaga
     6301 acagtaacag cagctggtgc agagaatatc caacaaaaaa cagatgagaa agtagatgaa
     6361 tcgggacctc ccgccccttc caaacccagg agaggacgtc gacccaagtc tgaatctcag
     6421 ggcaatgcta ccaaaaatga tgatctaaat aaacctatta acaagggaag gaagagagct
     6481 gcagtgggtc aggagagccc tgggggtttg gaagcaggta atgccaaagc acccaaactg
     6541 caagatttag ccaaaaaggc agcaccagca gaaagacaaa ttgacttaca aaggtaagtt
     6601 taacgcggcc gctcaggtca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc
     6661 aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac
     6721 atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca
     6781 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
     6841 ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct
     6901 gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc
     6961 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt
     7021 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt
     7081 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc
     7141 gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     7201 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     7261 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     7321 aaacctgtcg tgccagcgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc
     7381 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     7441 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     7501 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     7561 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     7621 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     7681 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     7741 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     7801 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     7861 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     7921 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     7981 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     8041 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     8101 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     8161 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     8221 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     8281 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     8341 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     8401 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     8461 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     8521 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     8581 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     8641 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     8701 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     8761 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     8821 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     8881 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     8941 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     9001 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     9061 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     9121 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     9181 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     9241 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     9301 ccgcgcacat ttccccgaaa agtgccacct g
//
