LOCUS Exported 9317 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION For transient expression of mouse PDS5A tagged with mKate2. ACCESSION . VERSION . KEYWORDS pKS016 - pCAGGS-3XFLAG-mKate2-Pds5a-bpA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9317) TITLE Molecular basis of CTCF binding polarity in genome folding JOURNAL Unpublished REFERENCE 2 (bases 1 to 9317) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9317 /organism="synthetic DNA construct" /mol_type="other DNA" 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(6676..6695) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 6741..6796 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(6795..6814) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(7157..7173) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7157..7173) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7170..7192) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7181..7197 /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(7205..7235) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7250..7271 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7396..7413) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7567..8155) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7647..7666) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8326..9186) /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 8949..8968 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9187..9291) /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 gsggggsggg 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 ggtatggact tcacgcagcc gaagcctgcc actgccctct gtggcgtcgt gagtgcggac 2641 gggaagatcg cttaccctcc gggagtaaaa gagatcaccg acaagatcac caccgatgaa 2701 atgatcaaac gtctgaagat ggtagtgaaa acttttatgg atatggatca ggattcagaa 2761 gatgaaaagc aacagtatct cccactagcc ttgcatctcg catctgaatt ctttctcagg 2821 aaccctaata aagatgtgcg ccttctcgta gcatgttgtt tggctgatat atttcgaatc 2881 tatgccccag aagctccata tacttcccat gataaactta aggacatatt tctctttatt 2941 accagacagt taaaaggttt ggaggataca aagagtccac agtttaatag atatttttat 3001 ttattagaga atttagcttg ggttaaatca tataacatct gctttgagtt ggaagattgc 3061 aatgaaattt ttattcagct ttttagaact ctcttctcgg taatcaacaa tagccacaat 3121 acgaaagtac aaatgcacat gttagacttg atgagttcta tcatcatgga aggtgatgga 3181 gttactcaag aattattgga ttccattctt atcaacctca tccctgcaca caagaactta 3241 aataaacagt ccttcgacct tgcaaaagtc ctactgaaaa ggacagtcca gactattgaa 3301 gcatgtattg ccaatttttt caatcaagtc ctggtgctgg gcagatcatc agtcagcgac 3361 ctgtctgaac acgtatttga tctgattcag gaactttttg ctatcgatcc tcagttactg 3421 ttatctgtca tgccacagct tgaattcaaa ctgaagagca acgatggtga agaacgccta 3481 gctgtggttc gactcctcgc aaaattgttc ggctctaaag attcagattt agcaacacag 3541 aatcggcctc tctggcagtg ctttcttggg cgatttaatg acattcatgt tcctgtgagg 3601 ttagaaagtg tgaagtttgc cagccactgt ttgatgaatc acccagactt agcgaaggat 3661 ctgacagaat atttgaaagt taggtcacat gatccagaag aagccattcg tcatgatgtt 3721 attgttacta taataacagc tgccaaaaga gaccttgcct tagtaaatga tcagttgctt 3781 ggctttgtca gggaaaggac actggataaa cggtggcgag taagaaaaga agccatgatg 3841 ggtctggctc agctctataa gaaatactgt cttcatgggg aagcaggaaa ggaagccgcg 3901 gagaaagtca gctggataaa ggacaagctg ctgcatatct actatcagaa cagcattgat 3961 gacaaactat tggtagagaa aatctttgct cagtatcttg ttccccacaa cctggaaaca 4021 gaagagagaa tgaaatgctt atattattta tatgctagct tggatccaaa tgctgtaaaa 4081 gctctcaatg aaatgtggaa atgtcagaac atgcttcgaa gtcatgtgcg tgaactattg 4141 gacttacaca agcagcctac atcagaagcg aactgttctg ccatgtttgg gaaactgatg 4201 accatagcaa agaatttgcc tgaccctgga aaagcacaag attttgtaaa gaaatttaac 4261 caggttcttg gtgatgatga gaaactgagg tctcagttgg aattattaat cagcccaacc 4321 tgttcatgca agcaggctga cgtttgtgtg agggaaatag ctcggaaact tgcaaatcct 4381 aagcagccaa ccaacccttt tctagagatg gtcaaatttc ttttggaaag aattgctcct 4441 gtgcacatcg attcagaagc cataagtgca ctggtaaaac tgatgaataa gtcaatcgaa 4501 gggacggccg atgatgaaga ggagggtgtc agtccagact cagccatccg ctcaggactt 4561 gagcttctta aggttctgtc tttcacacat cctacctcgt tccactctgc agagacatat 4621 gagtccttgt tacagtgcct aagaatggag gatgacaagg tagcagaagc tgcaatacaa 4681 atttttagaa acacaggcca caaaatagaa actgaccttc cccagatacg gtcaaccttg 4741 atccccattt tacatcagaa agccaagcgg ggcactccac accaagcaaa gcaggctgtt 4801 cactgcatcc atgccatctt ctcaaacaag gaggtccagc tggcacagat ttttgagcca 4861 ctcagtagga gtctgaatgc tgatgtacca gagcaactta taactccatt agtttcactg 4921 ggccacattt ccatgttagc accagatcag tttgcctccc cgatgaaatc tgtagtggca 4981 aactttattg ttaaagacct tctaatgaac gacaggtcaa caggtgagaa gaatggaaaa 5041 ttatggtctc cagatgagga agtttctcct gaagttctag caaaggtaca ggcaattaaa 5101 cttctggtaa ggtggctgtt gggtatgaaa aacaaccagt ctaaatctgc caactcaact 5161 cttcggttat tatcagccat gctggttagt gagggcgacc tgacagagca aaagaggatc 5221 agtaaatctg atatgtctcg cttgcgatta gctgctggta gtgccataat gaagcttgct 5281 caggaacctt gctaccatga aattattacc ccagaacagt ttcagctctg tgcactggtt 5341 attaatgatg agtgctacca agtaaggcag atatttgccc agaagcttca taaggcactt 5401 gtgaagttgc ttctcccact ggagtatatg gccatctttg ctttgtgtgc caaagaccct 5461 gtgaaagaaa ggagagcaca cgctcggcag tgtttgttaa agaacatcag catccgcagg 5521 gagtacatca aacagaaccc catggccact gagaaattat tgtcactgct gcctgaatat 5581 gtggttccat acatgattca cctcctagcc catgatcctg atttcacacg gtcacaagat 5641 gttgatcaac ttcgtgatat aaaagagtgc ctgtggttta tgcttgaagt cttaatgaca 5701 aagaatgaaa acaacagcca tgcattcatg aaaaagatgg cagagaatat caagctaacc 5761 agagacgccc agtctcccga tgaatccaag acaaatgaaa aactttatac ggtttgtgac 5821 gtggctctgt gtgttataaa tagtaaaagt gctttgtgca atgcagactc accaaaggac 5881 ccagtcctcc caatgaagtt ctttacacag cctgaaaagg acttctgtaa tgacaaaagc 5941 tatatttcag aagagacaag agttcttctg ttaacaggaa agccaaagcc tactggagta 6001 ttaggtacag tgaacaagcc tttatcagca acgggaagga agccttatgt tagaagcgcc 6061 ggcacagaga ctggaagcaa tattaacgcc aattcagagc tgagtccttc agccggaagt 6121 cgttcaaggg aacagagttc agaggcatca gaaactggag ttagtgaaaa tgaggagaat 6181 cctgtgagaa taatttctgt cacacctgta aagaatattg atactgtaaa gaataaggaa 6241 attaattctg atcagtctac ccaaggcaac atcagcagtg acagaggaaa gaaaagaatt 6301 gtaacagcag ctggtgcaga gaatatccaa aagccagacg agaaagtaga tgagtcagga 6361 ccgcctgccc cttccaaacc ccggagagga cgtcgcccca aatctgaatc tcagggcaat 6421 gcaaccaaaa acgatgatct aaataaacct gttagcaagg gaaggaaaag agctgcaggc 6481 agccaggaga gtctggaggc agggaatgcc aaagcgccca agctacaaga tggagccaaa 6541 aaggcagttc cagcagagag acaaattgat ttacaaaggt aagtttaaac gcggccgctc 6601 aggtcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca 6661 caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct 6721 tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa 6781 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa 6841 tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg 6901 ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca 6961 tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc 7021 tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg 7081 actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa 7141 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 7201 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 7261 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 7321 agcggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 7381 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 7441 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 7501 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 7561 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 7621 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 7681 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 7741 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 7801 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 7861 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 7921 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7981 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc 8041 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 8101 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 8161 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 8221 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 8281 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 8341 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 8401 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 8461 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 8521 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 8581 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 8641 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 8701 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 8761 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 8821 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 8881 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 8941 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 9001 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 9061 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 9121 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 9181 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 9241 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 9301 ccgaaaagtg ccacctg //