LOCUS       sequence-310236-        7205 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For transient expression of mouse BORIS (CTCFL) tagged with eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7205)
  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 7205)
  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: pKS031 - pCAGGS-3XFLAG-BORIS-eGFP-bpA
FEATURES             Location/Qualifiers
     source          1..7205
                     /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             join(3763..3765,3766..4476)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(3763..3765,3766..4476)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3805..3826)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4066..4085)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4413..4434
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4564..4583)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4629..4684
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4683..4702)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5045..5061)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5045..5061)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5058..5080)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5069..5085
                     /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(5093..5099,5100..5117,5118..5123))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5138..5159
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5284..5301)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5455..6043)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5535..5554)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6214..7005,7006..7074))
                     /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     6837..6856
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7075..7179)
                     /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 accggtgctg ccgctgaggt ccctgtccct
     1861 tctgggtact tcacccagat caaagagcag aagttgaagc ctggagacct agaggaggag
     1921 aaagaggagg acggggtaca aagagtggaa gcccaggagg gagttgtcaa ggaggtggag
     1981 gccgagaaca gttgcctgct tctggaggcc agggccccgg tggagagcga caggcggatc
     2041 ctgaccctgc aaacggtgca cctggagtcc caggatgtgc acctacaggg gctgggatgg
     2101 ctgagcgtgc cacactctga ggagctttca gggacggtac cagaggcgga aggcatactg
     2161 cagttgccat ccgtgctgtg gctcgaccca gagccccagc tcagccttca gcattgcgtg
     2221 acggtcagca tcccggaaga gctgtaccca ccagaggagc tgcagcggat acattttcac
     2281 ctgctgagag agaatgtgct aatggccgag gagaacccag agttaacacc agacttggac
     2341 gaaagcacag ccctgaaaaa gcccgaagaa gatgaaaagg accagctccc gccccaggga
     2401 gagacagaca agagagaaga gaggttgctc cttctggaaa tgaaaccaaa agagggaaaa
     2461 gacgacgaaa ttgtcctgac catttcccat ctaagcctcg aagaacagca agatccacca
     2521 gcggccaatc agacaagtgt gccgggagcc aaagccgcaa aaccaaaacg gcggaggcag
     2581 accaagggaa agcctcagag ctttcagtgt gacacctgcc cgttcacttc ctccaagctc
     2641 tcaactttca atcgtcacat caaaattcac agcaatgaga ggccacacct gtgtcacctg
     2701 tgcctgaagg ccttccggac tgtcactctt cttaggaacc atgtgaacac ccacacagga
     2761 accaggcccc acaagtgcag ggactgcgac atggcgtttg tcaccagtgg agaactcgtc
     2821 cggcacaggc gttacaaaca cacttatgag aagcccttca agtgctccct gtgcaagtac
     2881 gccagcgtcg aggcaagcaa gatgaagcgt cacatccgct cacacacggg tgagcgtccc
     2941 ttccagtgtt gccagtgtgc ttacgccagc agggactcct acaagctgaa gcgccacatg
     3001 aggacacact caggtgagaa gccgtatgaa tgtcccacct gtcacgtccg gttcacccag
     3061 agcgggacca tgaaaatcca tatagcacag aagcacggag agaatgtgcc caaatacgag
     3121 tgtccccact gtgccaccat catcgcgagg aagagcgacc tgcgtgtcca tctgcgtaac
     3181 ctgcacagcc agagcccgga ggagatgaag tgccgatact gtcccgctgg cttccatgag
     3241 cgctatgccc tcattcagca ccagaggacc cacaagaacg agaagaagtt caagtgcaag
     3301 cagtgcgatt acgcgtgcaa gcaggagcga tgcttgaagg cgcacatgcg catgcacaca
     3361 ggagagaagc ccttctcctg cctggcctgc aacaagcact tccgacagaa gcagctactg
     3421 accgtgcacc tgaggaagta ccatgacccg aacttcgtcc ccaatctgca cctgtgcctc
     3481 aagtgtgata aacgtttctc ccgctggagt aacctgcaga gacacagaaa gaagtgtgac
     3541 ccggagcatg agacgttagc ccccaacaag gacaggagac cagtgacaag gacacaggcc
     3601 tcggagggag aagcaggaca caaggaaggg gagcctcagt gccctgggga gcaggctctg
     3661 ggccaccaag gagaagcagc ggggagccag agcccagacc acggccttac ctgcgagata
     3721 atctttaaca tgatggataa gcttaagggg ggagcaggag gcgtgagcaa gggcgaggag
     3781 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     3841 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     3901 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     3961 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     4021 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     4081 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     4141 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     4201 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     4261 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     4321 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     4381 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     4441 gccgggatca ctctcggcat ggacgagctg tacaagtcgc gataagctag ctcaggtgca
     4501 ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg
     4561 agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac
     4621 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     4681 ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt
     4741 ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag
     4801 aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt
     4861 gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct
     4921 aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt
     4981 catagctgtc cctcttctct tatggagatc cctcgacctg cagcccaagc ttggcgtaat
     5041 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5101 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5161 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag cggatccgct
     5221 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5281 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5341 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5401 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5461 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5521 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5581 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5641 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5701 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5761 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5821 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5881 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5941 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6001 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6061 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6121 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6181 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6241 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6301 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6361 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6421 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6481 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6541 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6601 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6661 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6721 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6781 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6841 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6901 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6961 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7021 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7081 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7141 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7201 acctg
//
