LOCUS Exported 4869 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mouse N-terminal CTCF CRISPR knock-in donor vector with miniAID degron and HaloTag. ACCESSION . VERSION . KEYWORDS pLX-N-CTCF-mAID-Halo-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4869) AUTHORS Xie L, Dong P, Chen X, Hsieh TS, Banala S, De Marzio M, English BP, Qi Y, Jung SK, Kieffer-Kwon KR, Legant WR, Hansen AS, Schulmann A, Casellas R, Zhang B, Betzig E, Lavis LD, Chang HY, Tjian R, Liu Z TITLE 3D ATAC-PALM: super-resolution imaging of the accessible genome. JOURNAL Nat Methods. 2020 Apr;17(4):430-436. doi: 10.1038/s41592-020-0775-2. Epub 2020 Mar 16. PUBMED 32203384 REFERENCE 2 (bases 1 to 4869) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4869 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(29..51) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 151..170 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 364..386 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 378..395 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 379..395 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS 915..938 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 948..1838 /codon_start=1 /product="modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives" /label=HaloTag(R) /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL IGSEIARWLSTLEISG" CDS 1851..1871 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="EDLYFQS" primer_bind complement(2648..2664) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2648..2664) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2661..2683) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2672..2688 /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(2696..2726) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2741..2762 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2879..2896) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3050..3638) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3130..3149) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3809..4669) /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 4432..4451 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4670..4774) /gene="bla" /label=AmpR promoter primer_bind 4842..4860 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt caaaccaaaa ttaacattgt 421 agtgtttctc ttagactgaa cttctgcagt atatttaaca cctgctgtca ccagttttct 481 gttaattttg agtataatct agagtcttga tcagtttcag atttttttgt tttgattggc 541 aagatttata atataggtgg tgaatttttt tttcttccat caaagaggaa gatatctatt 601 aaataaccat tgatggatgc ctaattcatt cactaaggtg ggttttttgg ggggtcagga 661 ggttgagttt taggggtttg gcttggtttt tggttcttag agatagggtt tctctgtgca 721 ggccaggctg gcctcaaact cagaaatctg cctgcctgtg ggttctaagt gctgggatta 781 aaggtgtgtg caccactacc acctgccact caaatgattt tagtttaaaa tgtgtgcttt 841 gatttgagca aaccacatca taaataacat ctgtgttctc cttaataaag gaaggggaga 901 tggaaactag aatggattac aaggatgacg atgacaagct aggctccatg gcagaaatcg 961 gtactggctt tccattcgac ccccattatg tggaagtcct gggcgagcgc atgcactacg 1021 tcgatgttgg tccgcgcgat ggcacccctg tgctgttcct gcacggtaac ccgacctcct 1081 cctacgtgtg gcgcaacatc atcccgcatg ttgcaccgac ccatcgctgc attgctccag 1141 acctgatcgg tatgggcaaa tccgacaaac cagacctggg ttatttcttc gacgaccacg 1201 tccgcttcat ggatgccttc atcgaagccc tgggtctgga agaggtcgtc ctggtcattc 1261 acgactgggg ctccgctctg ggtttccact gggccaagcg caatccagag cgcgtcaaag 1321 gtattgcatt tatggagttc atccgcccta tcccgacctg ggacgaatgg ccagaatttg 1381 cccgcgagac cttccaggcc ttccgcacca ccgacgtcgg ccgcaagctg atcatcgatc 1441 agaacgtttt tatcgagggt acgctgccga tgggtgtcgt ccgcccgctg actgaagtcg 1501 agatggacca ttaccgcgag ccgttcctga atcctgttga ccgcgagcca ctgtggcgct 1561 tcccaaacga gctgccaatc gccggtgagc cagcgaacat cgtcgcgctg gtcgaagaat 1621 acatggactg gctgcaccag tcccctgtcc cgaagctgct gttctggggc accccaggcg 1681 ttctgatccc accggccgaa gccgctcgcc tggccaaaag cctgcctaac tgcaaggctg 1741 tggacatcgg cccgggtctg aatctgctgc aagaagacaa cccggacctg atcggcagcg 1801 agatcgcgcg ctggctgtcg acgctcgaga tttccggcga gccaaccact gaggatctgt 1861 actttcagag cgataactcc ggccgtacgg ggggagcagg aggcgtcgac ggtatcgatc 1921 ctaaagatcc agccaaacct ccggccaagg cacaagttgt gggatggcca ccggtgagat 1981 cataccggaa gaacgtgatg gtttcctgcc aaaaatcaag cggtggcccg gaggcggcgg 2041 cgttcgtgaa gaagcttgat ggtggaggtt ccggtgaagc tgtcgaggct atcgttgaag 2101 agtctgaaac tttcattaaa ggaaaagaaa gaaagacata tcagcgcaga agagagggag 2161 gacaggaaga agatgcttgc cacctgcccc agaaccagac agatgggggt gaggtggtcc 2221 aggatgtcaa cagcagtgta cagatggtaa tgatggaaca gctggatcct acccttctcc 2281 agatgaagac tgaagtcatg gagggtacag tggctccgga agcagaggct gcagtggacg 2341 atacccagat cataaccttg caggttgtaa atatggagga acagcccatt aacataggag 2401 agcttcagct tgtccaagta cctgttcctg tgacggtacc tgttgctact acttcagtag 2461 aagaacttca gggggcttat gagaatgaag tgtctaaaga gggccttgca gaaagtgaac 2521 cgatgatatg tcacacctta cctttgcctg aaggatttca ggtggtgaaa gtgggggcca 2581 atggagaagt ggaagatccc gggcccgtcg actgcagagg cctgcatgca agcttggcgt 2641 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 2701 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 2761 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 2821 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 2881 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 2941 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 3001 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 3061 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 3121 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 3181 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 3241 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 3301 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 3361 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 3421 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 3481 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 3541 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 3601 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 3661 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 3721 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 3781 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 3841 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 3901 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 3961 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 4021 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 4081 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 4141 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 4201 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 4261 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 4321 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 4381 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 4441 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 4501 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 4561 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 4621 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 4681 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 4741 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 4801 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc 4861 cctttcgtc //