LOCUS Exported 6330 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS XE257 pCDNA3.1(zeo)-hDsh3-truncDEP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6330) TITLE Xenopus Expression Plasmids REFERENCE 2 (bases 1 to 6330) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6330 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 935..944 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 944..967 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" protein_bind 1592..1603 /label=ZF binding site /bound_moiety="CCR5TC zinc-finger domain" /note="target sequence for the CCR5TC zinc-finger domain from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 2008; Gross et al., 2013)" primer_bind complement(2337..2354) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2343..2567 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2613..3041 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2700..2719) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2910..2931 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3050..3070) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3055..3385 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3236..3371 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3298..3317 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 3433..3480 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 3499..3873 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" polyA_signal 4003..4124 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4040..4059) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4094..4113 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4173..4189) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4173..4189) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4186..4208) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4197..4213 /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(4221..4251) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4266..4287 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4404..4421) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4575..5163) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4655..4674) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5334..6194) /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 5957..5976 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6195..6299) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggactaca aggacgacga 961 cgacaagggc ggcgcgcctg tacagattcg agagggtgga agtggaccag gtggtatggg 1021 cgagaccaag atcatctacc acttggatgg gcaggagacg ccgtaccttg tgaagctgcc 1081 cctgcccgcc gagcgcgtca ccttggcgga ctttaagggc gttttgcagc gacccagcta 1141 taagttcttc ttcaagtcta tggacgacga tttcggagtg gtgaaggagg agatctcgga 1201 tgacaatgcc aagctaccat gcttcaatgg ccgggtggtg tcctggctgg tgtcagctga 1261 gggctcacac ccagacccag cccccttctg tgctgataac ccatcggagc tgccaccacc 1321 tatggagcgc acgggaggca tcggggactc ccgaccccca tccttccacc ctcatgctgg 1381 tgggggcagc caggagaacc tggacaatga cacagagacg gactctttgg tgtctgccca 1441 gcgagagcgg ccacgccgga gggatggccc agagcatgca acccggctaa atggaactgc 1501 gaagggggaa cggcggcgag aaccaggggg ttatgatagc tcatccaccc ttatgagcag 1561 tgagctggag accaccagct tctttgactc agatgaggat gactccacca gcaggttcag 1621 cagctccaca gaacagagca gtgcctcacg cctgatgaga agacacaagc ggcggcggcg 1681 gaagcagaag gtttctcgga ttgagcggtc ctcgtccttc agcagcatca cggactccac 1741 catgtcactc aacatcatca cggtcactct caacatggaa aaatataact tcttgggcat 1801 ctccattgtg ggccaaagca acgagcgtgg tgacggcggc atctacattg gctctatcat 1861 gaagggtggg gccgtggctg ctgatggacg catcgagcca ggagatatgt tgttacaggt 1921 aaacgagatc aactttgaga acatgagtaa tgacgatgca gtccgggtac tgcgggagat 1981 tgtgcacaaa ccggggccca tcaccctgac tgtagccaag tgctgggacc caagtccacg 2041 tggttgcttc acattgccca ggagcgagcc catccggccc attgaccctg cggcctgggt 2101 ctcccacact gcagccatga ccggcacctt ccctgcatac ggcatgagcc cctccctgag 2161 caccatcacc tccaccagct cctccatcac cagttccatc cctgacacag agcgcctaga 2221 cgacttccac ttgtccatcc acagtgacat ggctgccatc gtaaaagcca tggcctcccc 2281 ttaggaattc gcccttttcg gatccactag agggcccgtt taaacccgct gatcagcctc 2341 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac 2401 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg 2461 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga 2521 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga 2581 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc 2641 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc 2701 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct 2761 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa 2821 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc 2881 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact 2941 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg 3001 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt 3061 cagttagggt gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc aaagcatgca 3121 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 3181 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc 3241 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 3301 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt 3361 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc 3421 tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac 3481 aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc 3541 gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg 3601 gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag 3661 gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg 3721 tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg 3781 accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac 3841 tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc 3901 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 3961 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct 4021 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 4081 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg 4141 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt 4201 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt 4261 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg 4321 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg 4381 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 4441 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 4501 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 4561 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 4621 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 4681 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 4741 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg 4801 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 4861 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 4921 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 4981 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg 5041 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 5101 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 5161 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 5221 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 5281 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 5341 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 5401 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 5461 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 5521 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 5581 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 5641 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 5701 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 5761 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 5821 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 5881 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 5941 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 6001 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 6061 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 6121 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 6181 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 6241 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 6301 acatttcccc gaaaagtgcc acctgacgtc //