LOCUS       sequence-310213-        9014 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 9014)
  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 9014)
  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..9014
                     /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(6373..6392)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6438..6493
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6492..6511)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6854..6870)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6854..6870)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6867..6889)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6878..6894
                     /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(6902..6908,6909..6926,6927..6932))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6947..6968
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7093..7110)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7264..7852)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7344..7363)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8023..8814,8815..8883))
                     /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     8646..8665
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8884..8988)
                     /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 gggggggggg 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 ggtatgatag cagctccaga aataccaact gattttaatc tactgcagga gtcagagaca
     2641 cacttttctt ctgacacaga ttttgaagac attgaaggaa aaaatcagaa gcaaggcaaa
     2701 ggcaaaactt gtaaaaaagg caaaaagggt ccagcagaaa agggtaaaag tggaaatgga
     2761 ggaggaaaac ctccttctgg ctcaaaccga atgaacggtc atcatcaaca gaatggagtt
     2821 gaaaacatga tgttgttcga agttgttaaa atgggcaaga gtgctatgca gtcggtggta
     2881 gatgactgga tcgagtccta caagcatgac cgggacatag cgcttcttga ccttatcaat
     2941 ttctttattc agtgctcagg ctgcaaagga gtcgtcacag cagagatgtt cagacatatg
     3001 cagaactctg agataattcg aaaaatgaca gaagaatttg acgaggatag tggggattac
     3061 ccacttacca tggctggtcc tcagtggaag aagttcaagt ccagcttctg tgagttcatt
     3121 ggtgtgttag tacggcagtg tcaatacagt atcatatatg atgagtacat gatggatact
     3181 gtcatttcac ttcttactgg attgtctgac tcccaagtca gagcatttcg acatacaagc
     3241 accctggcag ctatgaagtt gatgactgct ttggtaaatg tggcactaaa tcttagcatt
     3301 aatatggata acacacaaag acagtacgag gcagaacgga ataaaatgat tggaaaacga
     3361 gctaacgaaa ggctagaact cctgttacaa aagcgtaaag agcttcagga aaatcaagat
     3421 gagatagaaa atatgatgaa tgcaatattt aaaggagtgt ttgtacatag atacagggat
     3481 gcaatagctg aaatcagagc tatatgcatt gaagagattg gcatttggat gaagatgtat
     3541 agtgatgcct tccttaatga cagctattta aaatatgttg gttggactat gcatgataag
     3601 caaggtgaag tcagactcaa atgtcttact gctttacaag ggctttacta taacaaagaa
     3661 cttaattcca aactggaact ctttaccagt cgattcaagg atagaattgt gtctatgacc
     3721 cttgacaaag aatatgatgt tgcagtacaa gcaataaagt tactcactct tgttttacag
     3781 agtagtgaag aagttctaac tgcagaagat tgtgaaaatg tctatcatct ggtttattca
     3841 gctcaccggc cagtagcagt tgcagctgga gaatttctct acaaaaagct cttcagccgc
     3901 agagacccag aagaggatgg gctgatgaaa agaaggggaa ggcaaggccc gaatgccaac
     3961 cttgtgaaga ctttggtgtt tttctttttg gaaagtgagt tacacgagca tgcagcttac
     4021 cttgtggaca gcatgtggga ttgtgctact gagctcctga aagactggga atgtatgaat
     4081 agcttgttat tagaagagcc tcttagtgga gaggaagctc taacagacag acaagagagt
     4141 gctttgattg aaataatgct ttgtactatt agacaagcag ctgaatgtca tcctcccgtg
     4201 ggaagaggga caggaaaaag ggtgctgaca gcaaaggaga agaagacaca gttggatgac
     4261 agaacaagaa tcactgagct gtttgctgtg gcccttccgc agttattagc taaatactct
     4321 gtagatgcag aaaaggtgac caatttgttg cagttacctc agtactttga tttggagata
     4381 tacaccactg gacgattaga aaagcatttg gatgccttat tgcgccagat ccgaaatatt
     4441 gtagagaagc acacagacac agatgttttg gaagcatgtt ctaaaactta ccatgcatta
     4501 tgtaacgaag agttcaccat cttcaacaga gtagatattt caagaagtca gctgattgat
     4561 gaactggcag ataaatttaa ccggcttctt gaagattttc tgcaagaggg agaagaacct
     4621 gatgaagatg acgcatatca ggtattgtca acattgaaga ggatcactgc ttttcataat
     4681 gctcatgatc tttcaaagtg ggatttgttc gcttgtaatt acaaactctt gaaaaccgga
     4741 atcgaaaatg gagatatgcc tgagcagatt gttattcacg cactgcagtg tgctcactat
     4801 gtaatccttt ggcagcttgc taagatcact gaaagcacgt ctacaaagga ggacttactg
     4861 cgcttaaaga aacagatgag ggtattttgc cagatatgtc agcattactt gaccaacgtg
     4921 aatactactg ttaaagaaca ggccttcact attctgtgtg acattttgat gatcttcagt
     4981 caccagatta tgtctggagg gcgtgacatg ttagagccat tagtttatac tcctgattct
     5041 tcattgcagt ctgagttgct cagctttatt ttggatcatg tcttcattga acaggatgat
     5101 gatagtaata gtgcagatgg tcagcaagag gatgaagcca gcaaaattga agctttgcac
     5161 aagaggagaa atttacttgc agcattttgt aagctaattg tatataccgt ggtggagatg
     5221 aacacagctg cagatatctt caagcagtat atgaagtatt ataatgatta tggcgatatc
     5281 atcaaagaaa caatgagtaa aacaaggcag attgacaaaa ttcagtgtgc taagaccctt
     5341 attctcagtt tgcagcagct ttttaatgag atgatacaag agaatggcta taattttgat
     5401 agatcatcct caacatttag tggcataaaa gaacttgctc ggcgttttgc tttaactttt
     5461 ggacttgatc aattgaaaac cagagaagcc attgccatgc tccacaaaga tggtatagaa
     5521 tttgctttta aggagcctaa tccacaaggg gagagccatc cacctttaaa cttggcattt
     5581 cttgatattc tcagtgaatt ttcttctaaa ctgctccgac aggataaaag aacagtgtat
     5641 gtttatttgg aaaagttcat gacctttcag atgtcactcc gaagagaaga tgtatggctt
     5701 cctctgatgt cttaccgaaa ttctttgcta gctggtggtg atgatgatac tatgtcagtc
     5761 attagtggaa tgagcagtcg gggatcaact gtacggagta aaaaatcaaa accatctaca
     5821 ggaaagcgga aagtggttga aggcatgcaa ctagcactcc ctgaagaaag cagtagcagt
     5881 gacagtatgt ggttgagcag agagcaaaca ctgcacaccc ctgtcatgat gcagacacca
     5941 cagctcacct ccactattat gagagaaccg aaaaggttac ggcctgaaga tagcttcatg
     6001 agtgtctatc cgatgcaggc agaacaccat caaacgcctc tggattataa tcggcgtggt
     6061 acaagcctaa tggaagatga tgaagagcca attgttgaag atgttatgat gtcctcagaa
     6121 ggaaggattg aggatcttaa tgagggaatg gactttgaca ccatggatat agacttgcca
     6181 ccatcaaaga atagaagaga gagaacagaa ctgaagcccg atttctttga tccagcttca
     6241 attatggatg aatcagttct tggggtgtca atgttttaag tttaaacgcg gccgctcagg
     6301 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
     6361 ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     6421 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     6481 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     6541 gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg
     6601 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag
     6661 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt
     6721 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact
     6781 actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc agcccaagct
     6841 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     6901 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6961 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     7021 ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     7081 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     7141 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     7201 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     7261 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     7321 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     7381 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     7441 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7501 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7561 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7621 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7681 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     7741 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7801 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7861 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7921 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7981 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     8041 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     8101 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     8161 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     8221 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     8281 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     8341 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     8401 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     8461 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8521 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8581 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8641 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8701 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8761 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8821 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8881 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8941 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     9001 aaaagtgcca cctg
//
