LOCUS       sequence-310248-        9095 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 9095)
  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 9095)
  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..9095
                     /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(6454..6473)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6519..6574
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6573..6592)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6935..6951)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6935..6951)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6948..6970)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6959..6975
                     /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(6983..6989,6990..7007,7008..7013))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7028..7049
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7174..7191)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7345..7933)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7425..7444)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8104..8895,8896..8964))
                     /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     8727..8746
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8965..9069)
                     /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 ggggggsggg 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 ggtatgatta cttcagagtt accagtgtta caggattcaa ctaatgaaac tactgcccac
     2641 tctgatgctg gcagcgaact tgaagaaaca gaggtcaaag gaaaaagaaa aaggggtcgt
     2701 cctggccggc ctccatcaac aaataagaaa cctcgaaaat ctccaggaga aaagagcaga
     2761 attgaagctg gaattcgagg agcaggtcgt ggaagagcca atgggcaccc ccaacagaac
     2821 ggcgacgggg atcctgtcac cttatttgag gtggtgaagc tggggaagag tgcaatgcag
     2881 tccgtggtgg atgactggat tgaattatat aaacaagaca gggacatcgc acttctggat
     2941 ttaatcaact tttttatcca gtgttcagga tgtcgaggta cggtcagaat agagatgttt
     3001 cgaaatatgc agaacgcaga aataatcaga aaaatgactg aagaatttga tgaggacagt
     3061 ggtgactacc cccttacaat gcctggtcct cagtggaaaa aatttcgttc caatttttgt
     3121 gaatttattg gagtcctgat tcgacagtgt caatatagca taatttatga tgaatatatg
     3181 atggacaccg taatttccct tttgactggt ttgtcagact cccaagtcag agcttttagg
     3241 catacaagta cccttgctgc aatgaagctg atgactgctc tggtgaatgt tgctttaaac
     3301 ctcagtattc atcaagataa tacacagagg caatatgaag ctgagagaaa taaaatgatt
     3361 gggaagagag ccaatgaaag actggagtta ctacttcaga aacgtaaaga gctacaagaa
     3421 aatcaagatg aaattgaaaa tatgatgaac tctatcttta agggtatatt tgttcatcga
     3481 taccgtgatg caattgctga aattcgagcc atctgtattg aagaaattgg agtgtggatg
     3541 aaaatgtata gcgatgcctt cctaaatgac agttacttga aatatgtagg atggactctt
     3601 cacgataggc aaggggaagt cagactgaag tgtttgaaag ctctacaaag cctgtatacc
     3661 aatagagaat tattccccaa attggagcta tttacaaatc gattcaagga tcgcattgta
     3721 tcaatgactc ttgacaaaga atatgatgtt gctgtggaag caatccgatt ggttactctg
     3781 atccttcatg gcagtgaaga agctctttcc aacgaagact gtgaaaatgt ttaccatttg
     3841 gtgtattcag cacatcgccc tgttgctgtg gcagctggag agttcctaca caaaaagctg
     3901 ttcagcagac atgacccaca agcagaggaa gcgttagcga agaggagagg aaggaacagt
     3961 ccaaacggga acctcattag gatgcttgtt cttttctttc tggaaagtga gttacatgaa
     4021 catgcagcct acttggtgga cagcttgtgg gagagctctc aagaactgtt gaaagactgg
     4081 gaatgtatga cagagttact attagaagaa cctgttcaag gagaagaagc aatgtctgac
     4141 cgtcaagaga gtgctcttat agagctaatg gtctgtacaa ttcgtcaggc agctgaggca
     4201 catcctccag tgggaagggg tactggcaag agagtgctaa cagccaaaga aaggaaaact
     4261 caaattgatg ataggaacaa attaactgaa cattttatca ttacacttcc tatgttacta
     4321 tcaaagtatt ctgcagatgc tgagaaggta gcaaacttgc tgcaaattcc acagtatttt
     4381 gatctagaaa tctacagcac aggtcggatg gaaaagcacc tggatgcttt attaaaacaa
     4441 attaagtttg ttgtagagaa acatgtagaa tcagatgtcc tagaagcctg cagtaaaacc
     4501 tacagcattt tatgcagtga agagtatacc attcagaatc gagttgatat tgctcggagc
     4561 caactgattg atgaatttgt agatcgattc aatcattctg tagaagacct gttgcaagag
     4621 ggtgaagaag ctgacgatga tgatatctac aatgttcttt ccaccttaaa gcgtttaact
     4681 tcttttcaca atgctcatga tctcactaaa tgggatctat ttggtaattg ctacagacta
     4741 ttgaagactg gaattgaaca tggagctatg ccagaacaga tagtcgtaca agcactgcag
     4801 tgttcccatt actcaattct gtggcagttg gtgaaaatta ctgacggatc tccttccaaa
     4861 gaggatttgt tggtattgag gaaaacagtg aagtcttttc tggctgtttg ccaacagtgc
     4921 ctatctaatg ttaatactcc agttaaggaa caggctttca tgctgctctg tgatctttta
     4981 atgattttta gccaccaatt aatgacaggt ggcagagagg gccttcagcc tttggtgttt
     5041 aatccagata ctggactcca gtctgaactc ctcagttttg tgatggatca tgttttcatt
     5101 gaccaagatg aggaaaacca aagcatggag ggtgatgagg aagatgaagc taataaaatt
     5161 gaagccttac ataaaaggag gaacctgctt gctgccttca gcaaacttat catttatgat
     5221 attgttgaca tgcacgcagc tgcagacatc tttaaacact acatgaagta ttacaatgac
     5281 tatggtgata ttattaagga aacactgagt aagaccaggc agattgataa aattcagtgt
     5341 gctaagactc tcattctcag tctgcaacag ttgtttaatg aacttgttca agaacaaggt
     5401 cctaatctgg ataggacatc tgcccacgtc agtgggatta aagaactggc acgtcgtttt
     5461 gcccttacat ttggattgga ccagatcaag acccgagaag ctgttgccac acttcacaag
     5521 gatggaatag agtttgcctt taaataccag aatcagaaag gacaagaata cccaccccct
     5581 aatctggctt ttctagaagt actaagtgaa ttttcttcta aactccttcg acaggacaag
     5641 aaaactgttc attcgtatct tgaaaaattc ctcaccgagc agatgatgga aaggagggag
     5701 gatgtgtggc ttccactcat ctcgtataga aactcattag tcactggagg tgaagatgac
     5761 aggatgtctg tgaacagtgg aagtagcagc agtaaaacgt cctcagtaag gagtaagaaa
     5821 ggaagacccc cactgcacag aaaacgagta gaagatgaaa gtctggataa cacatggcta
     5881 aataggactg acacgatgat tcagactcct ggacccttgc caaccccaca gctcacctcc
     5941 acggtactta gagaaaacag ccgtcccatg ggagagcaga ttcaggaacc tgagtctgag
     6001 catggctctg aaccagactt tttacataat ccccagatgc agatctcttg gttaggccag
     6061 ccgaagttag aagacttgaa tcggaaggac agaacaggga tgaactacat gaaagtaaga
     6121 gctggagtcc ggcacgccgt tcggggtcta atggaggaag atgctgagcc catctttgaa
     6181 gatgtgatga tgtcatcacg gagccagtta gaagacatga atgaagaatt tgaagacacc
     6241 atggttattg atttgccccc atcaagaaat cgccgagaga gagctgagct aaggccagac
     6301 ttctttgact ctgcagctat catagaagat gattcaggat ttggaatgcc tatgttctga
     6361 gtttaaacgc ggccgctcag gtcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     6421 ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
     6481 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     6541 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     6601 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     6661 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     6721 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     6781 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     6841 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc
     6901 cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6961 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     7021 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     7081 cgggaaacct gtcgtgccag cggatccgct gcattaatga atcggccaac gcgcggggag
     7141 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     7201 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     7261 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     7321 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     7381 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7441 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7501 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7561 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7621 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7681 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7741 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     7801 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7861 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7921 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7981 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     8041 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     8101 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     8161 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     8221 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     8281 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     8341 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     8401 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     8461 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     8521 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8581 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8641 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8701 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8761 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8821 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8881 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8941 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     9001 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     9061 ggttccgcgc acatttcccc gaaaagtgcc acctg
//
