LOCUS Exported 8970 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION TALEColor-mVenus-Pan-Centromere. ACCESSION . VERSION . KEYWORDS pTH-PanCen-mVenus SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8970) AUTHORS Ma H, Reyes-Gutierrez P, Pederson T TITLE Visualization of repetitive DNA sequences in human chromosomes with transcription activator-like effectors. JOURNAL Proc Natl Acad Sci U S A. 2013 Dec 9. PUBMED 24324157 REFERENCE 2 (bases 1 to 8970) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8970 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(43..62) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 234..613 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 614..817 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 768..788 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 819..837 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 840..858 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 868..892 /label=LNCX /note="Human CMV promoter, forward primer" regulatory 1004..1013 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1013..1078 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 1085..1105 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 3644..4363 /codon_start=1 /product="Venus YFP with monomerizing A206K mutation (Nagai et al., 2002; Kremers et al., 2006)" /label=mVenus /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3689..3710) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3950..3969) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4297..4318 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(4395..4412) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4401..4625 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4671..5099 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4758..4777) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4968..4989 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5108..5128) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5113..5442 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5293..5428 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5355..5374 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5491..6516 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" polyA_signal 6646..6767 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6683..6702) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6737..6756 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6816..6832) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6816..6832) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6829..6851) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6840..6856 /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(6864..6894) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6909..6930 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7047..7064) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7218..7803) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7298..7317) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7974..8834) /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 8597..8616 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8835..8939) /gene="bla" /label=AmpR promoter ORIGIN 1 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc 61 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc 121 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattccatga agaatctgct 181 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg 241 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat 301 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc 361 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca 421 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta 481 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta 541 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat 601 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga 661 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca 721 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg 781 taggcgtgta cggtgggagg tctatataag cagagctctc cctatcagtg atagagatct 841 ccctatcagt gatagagatc gtcgacgagc tcgtttagtg aaccgtcaga tcgcctggag 901 acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca gcctggcgcg 961 ccggttctgg atccggttct caattgggtt ctaccggtta ggggccacca tggactataa 1021 ggaccacgac ggagactaca aggatcatga tattgattac aaagacgatg acgataagat 1081 ggccccaaag aagaagcgga aggtcggtat ccacggagtc ccagcagccg tagatttgag 1141 aactttggga tattcacagc agcagcagga aaagatcaag cccaaagtga ggtcgacagt 1201 cgcgcagcat cacgaagcgc tggtgggtca tgggtttaca catgcccaca tcgtagcctt 1261 gtcgcagcac cctgcagccc ttggcacggt cgccgtcaag taccaggaca tgattgcggc 1321 gttgccggaa gccacacatg aggcgatcgt cggtgtgggg aaacagtgga gcggagcccg 1381 agcgcttgag gccctgttga cggtcgcggg agagctgaga gggcctcccc ttcagctgga 1441 cacgggccag ttgctgaaga tcgcgaagcg gggaggagtc acggcggtcg aggcggtgca 1501 cgcgtggcgc aatgcgctca cgggagcacc cctcaacctg accccagagc aggtcgtggc 1561 aattgcgagc aacatcgggg gaaagcaggc actcgaaacc gtccagaggt tgctgcctgt 1621 gctgtgccaa gcgcacggac ttacgccaga gcaggtcgtg gcaattgcga gcaaccacgg 1681 gggaaagcag gcactcgaaa ccgtccagag gttgctgcct gtgctgtgcc aagcgcacgg 1741 actaacccca gagcaggtcg tggcaattgc gagcaacatc gggggaaagc aggcactcga 1801 aaccgtccag aggttgctgc ctgtgctgtg ccaagcgcac gggttgaccc cagagcaggt 1861 cgtggcaatt gcgagccatg acgggggaaa gcaggcactc gaaaccgtcc agaggttgct 1921 gcctgtgctg tgccaagcgc acggcctgac cccagagcag gtcgtggcaa ttgcgagcaa 1981 catcggggga aagcaggcac tcgaaaccgt ccagaggttg ctgcctgtgc tgtgccaagc 2041 gcacggactg acaccagagc aggtcgtggc aattgcgagc aaccacgggg gaaagcaggc 2101 actcgaaacc gtccagaggt tgctgcctgt gctgtgccaa gcgcacggac ttacacccga 2161 acaagtcgtg gcaattgcga gcaacatcgg gggaaagcag gcactcgaaa ccgtccagag 2221 gttgctgcct gtgctgtgcc aagcgcacgg acttacgcca gagcaggtcg tggcaattgc 2281 gagcaacatc gggggaaagc aggcactcga aaccgtccag aggttgctgc ctgtgctgtg 2341 ccaagcgcac ggactaaccc cagagcaggt cgtggcaatt gcgagcaacc acgggggaaa 2401 gcaggcactc gaaaccgtcc agaggttgct gcctgtgctg tgccaagcgc acgggttgac 2461 cccagagcag gtcgtggcaa ttgcgagcca tgacggggga aagcaggcac tcgaaaccgt 2521 ccagaggttg ctgcctgtgc tgtgccaagc gcacggcctg accccagagc aggtcgtggc 2581 aattgcgagc aacatcgggg gaaagcaggc actcgaaacc gtccagaggt tgctgcctgt 2641 gctgtgccaa gcgcacggac tgacaccaga gcaggtcgtg gcaattgcga gcaacggagg 2701 gggaaagcag gcactcgaaa ccgtccagag gttgctgcct gtgctgtgcc aagcgcacgg 2761 cctcacccca gagcaggtcg tggcaattgc gagcaacgga gggggaaagc aggcactcga 2821 aaccgtccag aggttgctgc ctgtgctgtg ccaagcgcac ggacttacgc cagagcaggt 2881 cgtggcaatt gcgagccatg acgggggaaa gcaggcactc gaaaccgtcc agaggttgct 2941 gcctgtgctg tgccaagcgc acggactaac cccagagcag gtcgtggcaa ttgcgagcaa 3001 cggaggggga aagcaggcac tcgaaaccgt ccagaggttg ctgcctgtgc tgtgccaagc 3061 gcacgggttg accccagagc aggtcgtggc aattgcgagc catgacgggg gaaagcaggc 3121 actcgaaacc gtccagaggt tgctgcctgt gctgtgccaa gcgcacggcc tgaccccaga 3181 gcaggtcgtg gcaattgcga gcaacatcgg gggaaagcag gcactcgaaa ccgtccagag 3241 gttgctgcct gtgctgtgcc aagcgcacgg actgacacca gagcaggtcg tggcaattgc 3301 gagcaaccac gggggaaagc aggcactcga aaccgtccag aggttgctgc ctgtgctgtg 3361 ccaagcgcac ggactcacgc ctgagcaggt agtggctatt gcatccaaca tcgggggcag 3421 acccgcactg gagtcaatcg tggcccagct ttcgaggccg gaccccgcgc tggccgcact 3481 cactaatgat catcttgtag cgctggcctg cctcggcgga cgacccgcct tggatgcggt 3541 gaagaagggg ctcccgcacg cgcctgcatt gattaagcgg accaacagaa ggattcccga 3601 gaggacatca catcgagtgg caggcctgca gggaagtgga agtatggtga gcaagggcga 3661 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca 3721 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa 3781 gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg 3841 ctacggcctg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa 3901 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa 3961 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct 4021 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta 4081 caacagccac aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt 4141 caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa 4201 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc 4261 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac 4321 cgccgccggg atcactctcg gcatggacga gctgtacaag tagtctagag ggcccgttta 4381 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc 4441 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga 4501 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca 4561 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc 4621 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg 4681 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc 4741 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg 4801 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg 4861 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg 4921 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt 4981 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt 5041 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta 5101 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga 5161 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc 5221 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc 5281 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 5341 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag 5401 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt 5461 atatccattt tcggatctga tcagcacgtg atgaaaaagc ctgaactcac cgcgacgtct 5521 gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag 5581 ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta 5641 aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc 5701 gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc 5761 atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct 5821 gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg 5881 agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc 5941 atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc 6001 agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa 6061 gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc 6121 ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc 6181 aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag 6241 cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt 6301 cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag 6361 ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc 6421 cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac 6481 cgacgcccca gcactcgtcc gagggcaaag gaatagcacg tgctacgaga tttcgattcc 6541 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg 6601 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca 6661 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 6721 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata 6781 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat 6841 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg 6901 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag 6961 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt 7021 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 7081 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 7141 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 7201 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 7261 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 7321 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 7381 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 7441 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 7501 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 7561 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 7621 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 7681 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 7741 accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 7801 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 7861 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 7921 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 7981 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 8041 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 8101 gcaatgatac cgcgagaacc acgctcaccg gctccagatt tatcagcaat aaaccagcca 8161 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 8221 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 8281 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 8341 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 8401 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 8461 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 8521 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 8581 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 8641 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 8701 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 8761 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 8821 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 8881 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 8941 acatttcccc gaaaagtgcc acctgacgtc //