LOCUS       sequence-301996-        8941 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For transient expression of mouse SMC3 tagged with mKate2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8941)
  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 8941)
  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: pKS037 - pCAGGS-3XFLAG-Smc3-mKate2-bpA
FEATURES             Location/Qualifiers
     source          1..8941
                     /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             5516..6211
                     /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(6302..6321)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6367..6422
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6421..6440)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6781..6797)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6781..6797)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6794..6816)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6805..6821
                     /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(6829..6835,6836..6853,6854..6859))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6874..6895
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7020..7037)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7191..7779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7271..7290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7950..8741,8742..8810))
                     /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     8573..8592
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8811..8915)
                     /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 agaccggtgg gggagcagga ggcatgtaca tcaagcaggt
     1861 gatcatccag ggcttccgaa gttaccgaga ccaaacaatt gtagatccct tcagctccaa
     1921 acataatgtc atcgtgggca gaaatggctc tggaaaaagc aacttctttt atgcaatcca
     1981 gtttgttctc agtgatgaat tcagccatct tcgtccagag cagcgattgg ctttgttgca
     2041 tgagggtaca ggtcctcgtg ttatttctgc ttttgtggaa atcatttttg acaattcgga
     2101 caaccggtta ccaattgata aagaggaagt ttcacttcga agagttattg gtgccaaaaa
     2161 ggatcagtat ttcctagata agaagatggt cacgaaaaat gatgtgatga atctccttga
     2221 aagtgctggg ttttccagaa gtaatcctta ttacattgtt aaacaaggaa agatcaacca
     2281 aatggcaacg gcaccagatt ctcagagatt aaagctgttg agagaagtag ctggtactag
     2341 agtgtatgat gaacggaaag aagaaagcat ctccctgatg aaagaaacag agggcaaacg
     2401 ggaaaagatc aatgagttgt tgaagtacat tgaagagcgc ctgcacactc tagaggagga
     2461 gaaggaggag ctggcccagt atcagaagtg ggataagatg aggcgcgccc tggagtacac
     2521 catctacaac caggagctca acgagacgcg cgctaagctc gacgagcttt ctgctaagcg
     2581 agaaacgagt ggagagaaat ccagacaact aagggatgcc cagcaggatg caagagacaa
     2641 aatggaggat attgagcgcc aggttagaga actgaaaaca aaaatttcag ccatgaaaga
     2701 agaaaaagaa cagcttagtg ctgaaagaca agagcagatt aagcaaagga ccaagttgga
     2761 gcttaaagcc aaggatttac aagatgagct ggcgggcaac agtgaacagc ggaaacgttt
     2821 attaaaagag aggcagaagc tgcttgaaaa aatagaagaa aagcagaaag aactggcaga
     2881 aacagaacct aaattcaaca gcgtaaaaga aaaagaagag cgaggaattg cgaggttggc
     2941 tcaagctaca caggaaagaa ctgatcttta tgcaaagcag ggtcgaggaa gccagtttac
     3001 gtcaaaagaa gaaagggata aatggattaa aaaggaactg aagtctttag atcaggcaat
     3061 taatgataag aaaagacaga ttgctgctat tcacaaggat ttggaggata ccgaggcaaa
     3121 taaagagaaa aatctggagc aatataataa attggatcag gatctcaatg aagtcaaagc
     3181 tcgagttgaa gaactggaca gaaaatatta tgaagtaaaa aataagaaag atgaactaca
     3241 aagtgaaaga aattacttgt ggagagagga gaatgcagaa caacaagcac ttgctgctaa
     3301 aagagaagac cttgagaaga agcagcaact tcttagagcg gcaacaggaa aggccatttt
     3361 aaatggaata gatagcatta acaaagtgct agaacatttt cggcgaaaag gtataaacca
     3421 gcatgttcaa aatggctacc atggcatcgt aatgaataac tttgagtgtg aaccagcttt
     3481 ctatacttgt gtggaagtca ctgctggtaa caggttattt tatcacattg ttgattcaga
     3541 tgaagtcagc acgaaaattt taatggagtt caataaaatg aatcttcctg gagaggtgac
     3601 tttcctgcct cttaacaagt tagatgtgag agatactgcc tatcctgaaa ctaacgatgc
     3661 tattcctatg atcagtaagc tgaggtacaa tcccagattt gacaaagctt tcaaacatgt
     3721 atttggaaag acactcatct gtcggagcat ggaagtttca actcagctgg cccgggcctt
     3781 cactatggac tgtattactt tggaaggtga tcaagtcagc catcgaggtg ctctgactgg
     3841 aggttattac gacacaagaa agtctcgact tgagttacag aaagacgtta ggaaggcaga
     3901 ggaggagctg ggtgagctgg aggctaagct caatgaaaac ctacgcagga acattgaaag
     3961 gattaataat gaaattgacc agttgatgaa ccaaatgcag cagatagaga cccaacaaag
     4021 aaaatttaaa gcatccagag atagcatatt atcagagatg aagatgctaa aagagaagag
     4081 acagcaatca gaaaagacct tcatgccaaa gcaacgtagc ttacaaagct tggaggcaag
     4141 tctgcatgct atggagtcca ccagagagtc actgaaagcg gagctaggaa cggatttgct
     4201 ttctcaactc agtctggaag atcagaaaag agtcgacgct ctgaatgatg aaatccgtca
     4261 gctgcagcag gaaaacagac agctgctaaa tgaaagaatt aaactagaag gcattattac
     4321 tcgagtagag acttacctga atgagaatct gaggaaacgc ttggaccaag tagaacagga
     4381 acttaatgaa ctgagagaga cagaaggtgg tactgttctt actgccacaa catcagaact
     4441 tgaagctatt aataaaagag taaaagatac tatggcaaga tcagaagatt tggacaattc
     4501 cattgacaaa acagaagctg gaattaaaga gctccagaaa agtatggagc gctggaaaaa
     4561 tatggagaaa gaacacatgg atgccataaa tcatgatact aaagagctgg agaagatgac
     4621 caaccggcaa ggcatgctgt tgaagaagaa ggaggagtgt atgaagaaga tccgggagct
     4681 ggggtccctc ccccaggaag cgtttgagaa gtaccagaca ctgagcctga agcagttgtt
     4741 tcgaaaactg gagcaatgca acacagagtt aaagaagtac agccacgtga acaaaaaggc
     4801 tctagatcag tttgtgaatt tctctgagca gaaggaaaag ctgataaagc ggcaagagga
     4861 attggataga ggctacaaat caatcatgga attgatgaat gtacttgaac ttagaaaata
     4921 tgaagctatt cagttaactt tcaaacaggt atctaagaac ttcagtgaag ttttccagaa
     4981 gttggtacct ggtggcaaag cgactttggt gatgaagaaa ggagatgtgg agggcagcca
     5041 gtctcaggac gaaggagaag ggagtggcga gagtgagcgg ggctctgggt cacagagcag
     5101 tgttccatca gttgaccagt tcacaggagt tgggatcagg gtgtcattta cagggaagca
     5161 aggggagatg agagaaatgc agcagctctc aggaggacag aaatctctgg tagcccttgc
     5221 tctcatcttt gccattcaga aatgcgaccc tgctcctttt tacctgtttg atgagatcga
     5281 ccaagctttg gatgctcagc acagaaaagc cgtgtcagat atgattatgg agctagccgt
     5341 gcatgcccag tttattacca ctactttcag gcctgaactg cttgagtcag ctgacaaatt
     5401 ctatggtgta aagttcagaa ataaggttag tcacattgat gtgatcacag cagagatggc
     5461 caaagatttt gtagaagacg ataccacgca tggtcttaag gggggagcag gaggcatggt
     5521 gagcgagctg attaaggaga acatgcacat gaagctgtac atggagggca ccgtgaacaa
     5581 ccaccacttc aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat
     5641 gagaatcaag gcggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctaccag
     5701 cttcatgtac ggcagcaaaa ccttcatcaa ccacacccag ggcatccccg acttctttaa
     5761 gcagtccttc cccgagggct tcacatggga gagagtcacc acatacgaag acgggggcgt
     5821 gctgaccgct acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaagat
     5881 cagaggggtg aacttcccat ccaacggccc tgtgatgcag aagaaaacac tcggctggga
     5941 ggcctccacc gagaccctgt accccgctga cggcggcctg gaaggcagag ccgacatggc
     6001 cctgaagctc gtgggcgggg gccacctgat ctgcaacttg aagaccacat acagatccaa
     6061 gaaacccgct aagaacctca agatgcccgg cgtctactat gtggacagaa gactggaaag
     6121 aatcaaggag gccgacaaag agacctacgt cgagcagcac gaggtggctg tggccagata
     6181 ctgcgacctc cctagcaaac tggggcacag aaagtcgcga taagctagct caggtgcagg
     6241 ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag
     6301 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     6361 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     6421 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt
     6481 agagtttggc aacatatgcc atatgctggc tgccatgaac aaaggtggct ataaagaggt
     6541 catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact
     6601 tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa
     6661 ttttccttac atgttttact agccagattt ttcctcctct cctgactact cccagtcata
     6721 gctgtccctc ttctcttatg aagatccctc gacctgcagc ccaagcttgg cgtaatcatg
     6781 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6841 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6901 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tccgctgcat
     6961 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     7021 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     7081 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     7141 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     7201 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     7261 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     7321 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     7381 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     7441 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7501 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7561 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7621 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7681 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7741 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7801 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7861 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7921 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7981 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     8041 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     8101 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     8161 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     8221 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     8281 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     8341 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     8401 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     8461 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     8521 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     8581 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     8641 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     8701 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     8761 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8821 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8881 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8941 g
//
