LOCUS       sequence-301989-        9085 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For transient expression of mouse SA1 (= STAG1 or SCC3A) tagged with
            mKate2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9085)
  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 9085)
  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: pKS021 - pCAGGS-3XFLAG-mKate2-SA1-bpA
FEATURES             Location/Qualifiers
     source          1..9085
                     /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(6446..6465)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6511..6566
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6565..6584)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6925..6941)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6925..6941)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6938..6960)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6949..6965
                     /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(6973..6979,6980..6997,6998..7003))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7018..7039
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7164..7181)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7335..7923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7415..7434)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8094..8885,8886..8954))
                     /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     8717..8736
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8955..9059)
                     /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 tacttcagag ttaccagtgt tacaggattc aactaatgaa actactgccc actctgatgc
     2641 tggcagcgaa cttgaagaaa cagaggtcaa aggaaaaaga aaaaggggtc gtcctggccg
     2701 gcctccatca acaaataaga aacctcgaaa atctccagga gaaaagagca gaattgaagc
     2761 tggaattcga ggagcaggtc gtggaagagc caatgggcac ccccaacaga acggcgacgg
     2821 ggatcctgtc accttatttg aggtggtgaa gctggggaag agtgcaatgc agtccgtggt
     2881 ggatgactgg attgaattat ataaacaaga cagggacatc gcacttctgg atttaatcaa
     2941 cttttttatc cagtgttcag gatgtcgagg tacggtcaga atagagatgt ttcgaaatat
     3001 gcagaacgca gaaataatca gaaaaatgac tgaagaattt gatgaggaca gtggtgacta
     3061 cccccttaca atgcctggtc ctcagtggaa aaaatttcgt tccaattttt gtgaatttat
     3121 tggagtcctg attcgacagt gtcaatatag cataatttat gatgaatata tgatggacac
     3181 cgtaatttcc cttttgactg gtttgtcaga ctcccaagtc agagctttta ggcatacaag
     3241 tacccttgct gcaatgaagc tgatgactgc tctggtgaat gttgctttaa acctcagtat
     3301 tcatcaagat aatacacaga ggcaatatga agctgagaga aataaaatga ttgggaagag
     3361 agccaatgaa agactggagt tactacttca gaaacgtaaa gagctacaag aaaatcaaga
     3421 tgaaattgaa aatatgatga actctatctt taagggtata tttgttcatc gataccgtga
     3481 tgcaattgct gaaattcgag ccatctgtat tgaagaaatt ggagtgtgga tgaaaatgta
     3541 tagcgatgcc ttcctaaatg acagttactt gaaatatgta ggatggactc ttcacgatag
     3601 gcaaggggaa gtcagactga agtgtttgaa agctctacaa agcctgtata ccaatagaga
     3661 attattcccc aaattggagc tatttacaaa tcgattcaag gatcgcattg tatcaatgac
     3721 tcttgacaaa gaatatgatg ttgctgtgga agcaatccga ttggttactc tgatccttca
     3781 tggcagtgaa gaagctcttt ccaacgaaga ctgtgaaaat gtttaccatt tggtgtattc
     3841 agcacatcgc cctgttgctg tggcagctgg agagttccta cacaaaaagc tgttcagcag
     3901 acatgaccca caagcagagg aagcgttagc gaagaggaga ggaaggaaca gtccaaacgg
     3961 gaacctcatt aggatgcttg ttcttttctt tctggaaagt gagttacatg aacatgcagc
     4021 ctacttggtg gacagcttgt gggagagctc tcaagaactg ttgaaagact gggaatgtat
     4081 gacagagtta ctattagaag aacctgttca aggagaagaa gcaatgtctg accgtcaaga
     4141 gagtgctctt atagagctaa tggtctgtac aattcgtcag gcagctgagg cacatcctcc
     4201 agtgggaagg ggtactggca agagagtgct aacagccaaa gaaaggaaaa ctcaaattga
     4261 tgataggaac aaattaactg aacattttat cattacactt cctatgttac tatcaaagta
     4321 ttctgcagat gctgagaagg tagcaaactt gctgcaaatt ccacagtatt ttgatctaga
     4381 aatctacagc acaggtcgga tggaaaagca cctggatgct ttattaaaac aaattaagtt
     4441 tgttgtagag aaacatgtag aatcagatgt cctagaagcc tgcagtaaaa cctacagcat
     4501 tttatgcagt gaagagtata ccattcagaa tcgagttgat attgctcgga gccaactgat
     4561 tgatgaattt gtagatcgat tcaatcattc tgtagaagac ctgttgcaag agggtgaaga
     4621 agctgacgat gatgatatct acaatgttct ttccacctta aagcgtttaa cttcttttca
     4681 caatgctcat gatctcacta aatgggatct atttggtaat tgctacagac tattgaagac
     4741 tggaattgaa catggagcta tgccagaaca gatagtcgta caagcactgc agtgttccca
     4801 ttactcaatt ctgtggcagt tggtgaaaat tactgacgga tctccttcca aagaggattt
     4861 gttggtattg aggaaaacag tgaagtcttt tctggctgtt tgccaacagt gcctatctaa
     4921 tgttaatact ccagttaagg aacaggcttt catgctgctc tgtgatcttt taatgatttt
     4981 tagccaccaa ttaatgacag gtggcagaga gggccttcag cctttggtgt ttaatccaga
     5041 tactggactc cagtctgaac tcctcagttt tgtgatggat catgttttca ttgaccaaga
     5101 tgaggaaaac caaagcatgg agggtgatga ggaagatgaa gctaataaaa ttgaagcctt
     5161 acataaaagg aggaacctgc ttgctgcctt cagcaaactt atcatttatg atattgttga
     5221 catgcacgca gctgcagaca tctttaaaca ctacatgaag tattacaatg actatggtga
     5281 tattattaag gaaacactga gtaagaccag gcagattgat aaaattcagt gtgctaagac
     5341 tctcattctc agtctgcaac agttgtttaa tgaacttgtt caagaacaag gtcctaatct
     5401 ggataggaca tctgcccacg tcagtgggat taaagaactg gcacgtcgtt ttgcccttac
     5461 atttggattg gaccagatca agacccgaga agctgttgcc acacttcaca aggatggaat
     5521 agagtttgcc tttaaatacc agaatcagaa aggacaagaa tacccacccc ctaatctggc
     5581 ttttctagaa gtactaagtg aattttcttc taaactcctt cgacaggaca agaaaactgt
     5641 tcattcgtat cttgaaaaat tcctcaccga gcagatgatg gaaaggaggg aggatgtgtg
     5701 gcttccactc atctcgtata gaaactcatt agtcactgga ggtgaagatg acaggatgtc
     5761 tgtgaacagt ggaagtagca gcagtaaaac gtcctcagta aggagtaaga aaggaagacc
     5821 cccactgcac agaaaacgag tagaagatga aagtctggat aacacatggc taaataggac
     5881 tgacacgatg attcagactc ctggaccctt gccaacccca cagctcacct ccacggtact
     5941 tagagaaaac agccgtccca tgggagagca gattcaggaa cctgagtctg agcatggctc
     6001 tgaaccagac tttttacata atccccagat gcagatctct tggttaggcc agccgaagtt
     6061 agaagacttg aatcggaagg acagaacagg gatgaactac atgaaagtaa gagctggagt
     6121 ccggcacgcc gttcggggtc taatggagga agatgctgag cccatctttg aagatgtgat
     6181 gatgtcatca cggagccagt tagaagacat gaatgaagaa tttgaagaca ccatggttat
     6241 tgatttgccc ccatcaagaa atcgccgaga gagagctgag ctaaggccag acttctttga
     6301 ctctgcagct atcatagaag atgattcagg atttggaatg cctatgttct gagtttaaac
     6361 gcggccgctc aggtcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg
     6421 ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca
     6481 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag
     6541 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa
     6601 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgccatatgc tggctgccat
     6661 gaacaaaggt ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc
     6721 ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta
     6781 tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc
     6841 ctctcctgac tactcccagt catagctgtc cctcttctct tatgaagatc cctcgacctg
     6901 cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6961 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     7021 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     7081 gtcgtgccag cggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg
     7141 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     7201 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     7261 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7321 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7381 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7441 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7501 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     7561 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7621 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7681 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7741 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     7801 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7861 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7921 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7981 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     8041 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     8101 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     8161 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     8221 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     8281 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     8341 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     8401 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     8461 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     8521 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8581 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8641 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8701 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8761 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8821 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8881 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8941 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     9001 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     9061 acatttcccc gaaaagtgcc acctg
//
