LOCUS       sequence-301990-        9004 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For transient expression of mouse SA2 (STAG2 or SCC3B) tagged with 
            mKate2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9004)
  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 9004)
  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: pKS022 - pCAGGS-3XFLAG-mKate2-SA2-bpA
FEATURES             Location/Qualifiers
     source          1..9004
                     /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(6365..6384)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6430..6485
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6484..6503)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6844..6860)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6844..6860)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6857..6879)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6868..6884
                     /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(6892..6898,6899..6916,6917..6922))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6937..6958
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7083..7100)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7254..7842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7334..7353)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8013..8804,8805..8873))
                     /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     8636..8655
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8874..8978)
                     /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 ccggtatgat
     2581 agcagctcca gaaataccaa ctgattttaa tctactgcag gagtcagaga cacacttttc
     2641 ttctgacaca gattttgaag acattgaagg aaaaaatcag aagcaaggca aaggcaaaac
     2701 ttgtaaaaaa ggcaaaaagg gtccagcaga aaagggtaaa agtggaaatg gaggaggaaa
     2761 acctccttct ggctcaaacc gaatgaacgg tcatcatcaa cagaatggag ttgaaaacat
     2821 gatgttgttc gaagttgtta aaatgggcaa gagtgctatg cagtcggtgg tagatgactg
     2881 gatcgagtcc tacaagcatg accgggacat agcgcttctt gaccttatca atttctttat
     2941 tcagtgctca ggctgcaaag gagtcgtcac agcagagatg ttcagacata tgcagaactc
     3001 tgagataatt cgaaaaatga cagaagaatt tgacgaggat agtggggatt acccacttac
     3061 catggctggt cctcagtgga agaagttcaa gtccagcttc tgtgagttca ttggtgtgtt
     3121 agtacggcag tgtcaataca gtatcatata tgatgagtac atgatggata ctgtcatttc
     3181 acttcttact ggattgtctg actcccaagt cagagcattt cgacatacaa gcaccctggc
     3241 agctatgaag ttgatgactg ctttggtaaa tgtggcacta aatcttagca ttaatatgga
     3301 taacacacaa agacagtacg aggcagaacg gaataaaatg attggaaaac gagctaacga
     3361 aaggctagaa ctcctgttac aaaagcgtaa agagcttcag gaaaatcaag atgagataga
     3421 aaatatgatg aatgcaatat ttaaaggagt gtttgtacat agatacaggg atgcaatagc
     3481 tgaaatcaga gctatatgca ttgaagagat tggcatttgg atgaagatgt atagtgatgc
     3541 cttccttaat gacagctatt taaaatatgt tggttggact atgcatgata agcaaggtga
     3601 agtcagactc aaatgtctta ctgctttaca agggctttac tataacaaag aacttaattc
     3661 caaactggaa ctctttacca gtcgattcaa ggatagaatt gtgtctatga cccttgacaa
     3721 agaatatgat gttgcagtac aagcaataaa gttactcact cttgttttac agagtagtga
     3781 agaagttcta actgcagaag attgtgaaaa tgtctatcat ctggtttatt cagctcaccg
     3841 gccagtagca gttgcagctg gagaatttct ctacaaaaag ctcttcagcc gcagagaccc
     3901 agaagaggat gggctgatga aaagaagggg aaggcaaggc ccgaatgcca accttgtgaa
     3961 gactttggtg tttttctttt tggaaagtga gttacacgag catgcagctt accttgtgga
     4021 cagcatgtgg gattgtgcta ctgagctcct gaaagactgg gaatgtatga atagcttgtt
     4081 attagaagag cctcttagtg gagaggaagc tctaacagac agacaagaga gtgctttgat
     4141 tgaaataatg ctttgtacta ttagacaagc agctgaatgt catcctcccg tgggaagagg
     4201 gacaggaaaa agggtgctga cagcaaagga gaagaagaca cagttggatg acagaacaag
     4261 aatcactgag ctgtttgctg tggcccttcc gcagttatta gctaaatact ctgtagatgc
     4321 agaaaaggtg accaatttgt tgcagttacc tcagtacttt gatttggaga tatacaccac
     4381 tggacgatta gaaaagcatt tggatgcctt attgcgccag atccgaaata ttgtagagaa
     4441 gcacacagac acagatgttt tggaagcatg ttctaaaact taccatgcat tatgtaacga
     4501 agagttcacc atcttcaaca gagtagatat ttcaagaagt cagctgattg atgaactggc
     4561 agataaattt aaccggcttc ttgaagattt tctgcaagag ggagaagaac ctgatgaaga
     4621 tgacgcatat caggtattgt caacattgaa gaggatcact gcttttcata atgctcatga
     4681 tctttcaaag tgggatttgt tcgcttgtaa ttacaaactc ttgaaaaccg gaatcgaaaa
     4741 tggagatatg cctgagcaga ttgttattca cgcactgcag tgtgctcact atgtaatcct
     4801 ttggcagctt gctaagatca ctgaaagcac gtctacaaag gaggacttac tgcgcttaaa
     4861 gaaacagatg agggtatttt gccagatatg tcagcattac ttgaccaacg tgaatactac
     4921 tgttaaagaa caggccttca ctattctgtg tgacattttg atgatcttca gtcaccagat
     4981 tatgtctgga gggcgtgaca tgttagagcc attagtttat actcctgatt cttcattgca
     5041 gtctgagttg ctcagcttta ttttggatca tgtcttcatt gaacaggatg atgatagtaa
     5101 tagtgcagat ggtcagcaag aggatgaagc cagcaaaatt gaagctttgc acaagaggag
     5161 aaatttactt gcagcatttt gtaagctaat tgtatatacc gtggtggaga tgaacacagc
     5221 tgcagatatc ttcaagcagt atatgaagta ttataatgat tatggcgata tcatcaaaga
     5281 aacaatgagt aaaacaaggc agattgacaa aattcagtgt gctaagaccc ttattctcag
     5341 tttgcagcag ctttttaatg agatgataca agagaatggc tataattttg atagatcatc
     5401 ctcaacattt agtggcataa aagaacttgc tcggcgtttt gctttaactt ttggacttga
     5461 tcaattgaaa accagagaag ccattgccat gctccacaaa gatggtatag aatttgcttt
     5521 taaggagcct aatccacaag gggagagcca tccaccttta aacttggcat ttcttgatat
     5581 tctcagtgaa ttttcttcta aactgctccg acaggataaa agaacagtgt atgtttattt
     5641 ggaaaagttc atgacctttc agatgtcact ccgaagagaa gatgtatggc ttcctctgat
     5701 gtcttaccga aattctttgc tagctggtgg tgatgatgat actatgtcag tcattagtgg
     5761 aatgagcagt cggggatcaa ctgtacggag taaaaaatca aaaccatcta caggaaagcg
     5821 gaaagtggtt gaaggcatgc aactagcact ccctgaagaa agcagtagca gtgacagtat
     5881 gtggttgagc agagagcaaa cactgcacac ccctgtcatg atgcagacac cacagctcac
     5941 ctccactatt atgagagaac cgaaaaggtt acggcctgaa gatagcttca tgagtgtcta
     6001 tccgatgcag gcagaacacc atcaaacgcc tctggattat aatcggcgtg gtacaagcct
     6061 aatggaagat gatgaagagc caattgttga agatgttatg atgtcctcag aaggaaggat
     6121 tgaggatctt aatgagggaa tggactttga caccatggat atagacttgc caccatcaaa
     6181 gaatagaaga gagagaacag aactgaagcc cgatttcttt gatccagctt caattatgga
     6241 tgaatcagtt cttggggtgt caatgtttta agtttaaacg cggccgctca ggtcaggtgc
     6301 aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact
     6361 gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga
     6421 cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc
     6481 tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg
     6541 tttagagttt ggcaacatat gccatatgct ggctgccatg aacaaaggtg gctataaaga
     6601 ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg
     6661 acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta
     6721 aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc
     6781 atagctgtcc ctcttctctt atgaagatcc ctcgacctgc agcccaagct tggcgtaatc
     6841 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6901 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6961 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgctg
     7021 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     7081 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     7141 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     7201 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     7261 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7321 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7381 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7441 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7501 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7561 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7621 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7681 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7741 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7801 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7861 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7921 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7981 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     8041 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     8101 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     8161 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     8221 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     8281 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     8341 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     8401 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     8461 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     8521 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8581 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     8641 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8701 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8761 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     8821 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8881 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8941 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     9001 cctg
//
