LOCUS       sequence-310250-        8951 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 8951)
  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 8951)
  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..8951
                     /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             5524..6219
                     /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(6310..6329)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6375..6430
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6429..6448)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6791..6807)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6791..6807)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6804..6826)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6815..6831
                     /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(6839..6845,6846..6863,6864..6869))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6884..6905
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7030..7047)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7201..7789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7281..7300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7960..8751,8752..8820))
                     /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     8583..8602
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8821..8925)
                     /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 rgggctcggg 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 accggtgggg gagcaggagg catgtacatc
     1861 aagcaggtga tcatccaggg cttccgaagt taccgagacc aaacaattgt agatcccttc
     1921 agctccaaac ataatgtcat cgtgggcaga aatggctctg gaaaaagcaa cttcttttat
     1981 gcaatccagt ttgttctcag tgatgaattc agccatcttc gtccagagca gcgattggct
     2041 ttgttgcatg agggtacagg tcctcgtgtt atttctgctt ttgtggaaat catttttgac
     2101 aattcggaca accggttacc aattgataaa gaggaagttt cacttcgaag agttattggt
     2161 gccaaaaagg atcagtattt cctagataag aagatggtca cgaaaaatga tgtgatgaat
     2221 ctccttgaaa gtgctgggtt ttccagaagt aatccttatt acattgttaa acaaggaaag
     2281 atcaaccaaa tggcaacggc accagattct cagagattaa agctgttgag agaagtagct
     2341 ggtactagag tgtatgatga acggaaagaa gaaagcatct ccctgatgaa agaaacagag
     2401 ggcaaacggg aaaagatcaa tgagttgttg aagtacattg aagagcgcct gcacactcta
     2461 gaggaggaga aggaggagct ggcccagtat cagaagtggg ataagatgag gcgcgccctg
     2521 gagtacacca tctacaacca ggagctcaac gagacgcgcg ctaagctcga cgagctttct
     2581 gctaagcgag aaacgagtgg agagaaatcc agacaactaa gggatgccca gcaggatgca
     2641 agagacaaaa tggaggatat tgagcgccag gttagagaac tgaaaacaaa aatttcagcc
     2701 atgaaagaag aaaaagaaca gcttagtgct gaaagacaag agcagattaa gcaaaggacc
     2761 aagttggagc ttaaagccaa ggatttacaa gatgagctgg cgggcaacag tgaacagcgg
     2821 aaacgtttat taaaagagag gcagaagctg cttgaaaaaa tagaagaaaa gcagaaagaa
     2881 ctggcagaaa cagaacctaa attcaacagc gtaaaagaaa aagaagagcg aggaattgcg
     2941 aggttggctc aagctacaca ggaaagaact gatctttatg caaagcaggg tcgaggaagc
     3001 cagtttacgt caaaagaaga aagggataaa tggattaaaa aggaactgaa gtctttagat
     3061 caggcaatta atgataagaa aagacagatt gctgctattc acaaggattt ggaggatacc
     3121 gaggcaaata aagagaaaaa tctggagcaa tataataaat tggatcagga tctcaatgaa
     3181 gtcaaagctc gagttgaaga actggacaga aaatattatg aagtaaaaaa taagaaagat
     3241 gaactacaaa gtgaaagaaa ttacttgtgg agagaggaga atgcagaaca acaagcactt
     3301 gctgctaaaa gagaagacct tgagaagaag cagcaacttc ttagagcggc aacaggaaag
     3361 gccattttaa atggaataga tagcattaac aaagtgctag aacattttcg gcgaaaaggt
     3421 ataaaccagc atgttcaaaa tggctaccat ggcatcgtaa tgaataactt tgagtgtgaa
     3481 ccagctttct atacttgtgt ggaagtcact gctggtaaca ggttatttta tcacattgtt
     3541 gattcagatg aagtcagcac gaaaatttta atggagttca ataaaatgaa tcttcctgga
     3601 gaggtgactt tcctgcctct taacaagtta gatgtgagag atactgccta tcctgaaact
     3661 aacgatgcta ttcctatgat cagtaagctg aggtacaatc ccagatttga caaagctttc
     3721 aaacatgtat ttggaaagac actcatctgt cggagcatgg aagtttcaac tcagctggcc
     3781 cgggccttca ctatggactg tattactttg gaaggtgatc aagtcagcca tcgaggtgct
     3841 ctgactggag gttattacga cacaagaaag tctcgacttg agttacagaa agacgttagg
     3901 aaggcagagg aggagctggg tgagctggag gctaagctca atgaaaacct acgcaggaac
     3961 attgaaagga ttaataatga aattgaccag ttgatgaacc aaatgcagca gatagagacc
     4021 caacaaagaa aatttaaagc atccagagat agcatattat cagagatgaa gatgctaaaa
     4081 gagaagagac agcaatcaga aaagaccttc atgccaaagc aacgtagctt acaaagcttg
     4141 gaggcaagtc tgcatgctat ggagtccacc agagagtcac tgaaagcgga gctaggaacg
     4201 gatttgcttt ctcaactcag tctggaagat cagaaaagag tcgacgctct gaatgatgaa
     4261 atccgtcagc tgcagcagga aaacagacag ctgctaaatg aaagaattaa actagaaggc
     4321 attattactc gagtagagac ttacctgaat gagaatctga ggaaacgctt ggaccaagta
     4381 gaacaggaac ttaatgaact gagagagaca gaaggtggta ctgttcttac tgccacaaca
     4441 tcagaacttg aagctattaa taaaagagta aaagatacta tggcaagatc agaagatttg
     4501 gacaattcca ttgacaaaac agaagctgga attaaagagc tccagaaaag tatggagcgc
     4561 tggaaaaata tggagaaaga acacatggat gccataaatc atgatactaa agagctggag
     4621 aagatgacca accggcaagg catgctgttg aagaagaagg aggagtgtat gaagaagatc
     4681 cgggagctgg ggtccctccc ccaggaagcg tttgagaagt accagacact gagcctgaag
     4741 cagttgtttc gaaaactgga gcaatgcaac acagagttaa agaagtacag ccacgtgaac
     4801 aaaaaggctc tagatcagtt tgtgaatttc tctgagcaga aggaaaagct gataaagcgg
     4861 caagaggaat tggatagagg ctacaaatca atcatggaat tgatgaatgt acttgaactt
     4921 agaaaatatg aagctattca gttaactttc aaacaggtat ctaagaactt cagtgaagtt
     4981 ttccagaagt tggtacctgg tggcaaagcg actttggtga tgaagaaagg agatgtggag
     5041 ggcagccagt ctcaggacga aggagaaggg agtggcgaga gtgagcgggg ctctgggtca
     5101 cagagcagtg ttccatcagt tgaccagttc acaggagttg ggatcagggt gtcatttaca
     5161 gggaagcaag gggagatgag agaaatgcag cagctctcag gaggacagaa atctctggta
     5221 gcccttgctc tcatctttgc cattcagaaa tgcgaccctg ctccttttta cctgtttgat
     5281 gagatcgacc aagctttgga tgctcagcac agaaaagccg tgtcagatat gattatggag
     5341 ctagccgtgc atgcccagtt tattaccact actttcaggc ctgaactgct tgagtcagct
     5401 gacaaattct atggtgtaaa gttcagaaat aaggttagtc acattgatgt gatcacagca
     5461 gagatggcca aagattttgt agaagacgat accacgcatg gtcttaaggg gggagcagga
     5521 ggcatggtga gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc
     5581 gtgaacaacc accacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
     5641 cagaccatga gaatcaaggc ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
     5701 gctaccagct tcatgtacgg cagcaaaacc ttcatcaacc acacccaggg catccccgac
     5761 ttctttaagc agtccttccc cgagggcttc acatgggaga gagtcaccac atacgaagac
     5821 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac
     5881 gtcaagatca gaggggtgaa cttcccatcc aacggccctg tgatgcagaa gaaaacactc
     5941 ggctgggagg cctccaccga gaccctgtac cccgctgacg gcggcctgga aggcagagcc
     6001 gacatggccc tgaagctcgt gggcgggggc cacctgatct gcaacttgaa gaccacatac
     6061 agatccaaga aacccgctaa gaacctcaag atgcccggcg tctactatgt ggacagaaga
     6121 ctggaaagaa tcaaggaggc cgacaaagag acctacgtcg agcagcacga ggtggctgtg
     6181 gccagatact gcgacctccc tagcaaactg gggcacagaa agtcgcgata agctagctca
     6241 ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata
     6301 ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca
     6361 tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     6421 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta
     6481 tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct
     6541 ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa
     6601 agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac
     6661 atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct cctgactact
     6721 cccagtcata gctgtccctc ttctcttatg gagatccctc gacctgcagc ccaagcttgg
     6781 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6841 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     6901 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga
     6961 tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     7021 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     7081 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     7141 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7201 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7261 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7321 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7381 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7441 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7501 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7561 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7621 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     7681 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7741 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7801 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7861 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7921 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7981 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     8041 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     8101 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     8161 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8221 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     8281 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8341 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8401 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8461 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8521 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8581 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8641 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8701 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8761 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8821 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8881 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8941 agtgccacct g
//
