LOCUS Exported 6922 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS XE253 pCDNA3.1(zeo)-hDsh3-82-716 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6922) TITLE Xenopus Expression Plasmids REFERENCE 2 (bases 1 to 6922) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6922 /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 1310..1321 /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(2929..2946) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2935..3159 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3205..3633 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3292..3311) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3502..3523 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3642..3662) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3647..3977 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3828..3963 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3890..3909 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 4025..4072 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 4091..4465 /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 4595..4716 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4632..4651) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4686..4705 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4765..4781) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4765..4781) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4778..4800) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4789..4805 /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(4813..4843) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4858..4879 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4996..5013) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5167..5755) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5247..5266) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5926..6786) /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 6549..6568 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6787..6891) /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 ggcgcgccag gctcacaccc agacccagcc cccttctgtg ctgataaccc 1021 atcggagctg ccaccaccta tggagcgcac gggaggcatc ggggactccc gacccccatc 1081 cttccaccct catgctggtg ggggcagcca ggagaacctg gacaatgaca cagagacgga 1141 ctctttggtg tctgcccagc gagagcggcc acgccggagg gatggcccag agcatgcaac 1201 ccggctaaat ggaactgcga agggggaacg gcggcgagaa ccagggggtt atgatagctc 1261 atccaccctt atgagcagtg agctggagac caccagcttc tttgactcag atgaggatga 1321 ctccaccagc aggttcagca gctccacaga acagagcagt gcctcacgcc tgatgagaag 1381 acacaagcgg cggcggcgga agcagaaggt ttctcggatt gagcggtcct cgtccttcag 1441 cagcatcacg gactccacca tgtcactcaa catcatcacg gtcactctca acatggaaaa 1501 atataacttc ttgggcatct ccattgtggg ccaaagcaac gagcgtggtg acggcggcat 1561 ctacattggc tctatcatga agggtggggc cgtggctgct gatggacgca tcgagccagg 1621 agatatgttg ttacaggtaa acgagatcaa ctttgagaac atgagtaatg acgatgcagt 1681 ccgggtactg cgggagattg tgcacaaacc ggggcccatc accctgactg tagccaagtg 1741 ctgggaccca agtccacgtg gttgcttcac attgcccagg agcgagccca tccggcccat 1801 tgaccctgcg gcctgggtct cccacactgc agccatgacc ggcaccttcc ctgcatacgg 1861 catgagcccc tccctgagca ccatcacctc caccagctcc tccatcacca gttccatccc 1921 tgacacagag cgcctagacg acttccactt gtccatccac agtgacatgg ctgccatcgt 1981 aaaagccatg gcctcccctg aatcagggtt ggaggtccgt gaccgcatgt ggctcaagat 2041 taccatccct aatgctttca tcggctcaga tgtggtggac tggctgtacc acaatgtgga 2101 aggcttcacg gaccggaggg aggcccgcaa gtatgccagc aacctgctga aagctggctt 2161 catccgccat accgtcaaca agatcacctt ctccgagcag tgctactaca tcttcggtga 2221 cctctgcggc aacatggcca acctgtctct ccacgatcac gatggctcca gtggcgcctc 2281 tgaccaggac acactggccc ctttgccgca cccgggggcc gccccttggc ccatggcttt 2341 cccgtaccag tacccgccac ccccgcaccc atacaacccg cacccgggct tcccggagct 2401 gggctacagc tacggcgggg gcagcgccag cagtcagcac agcgaaggca gtcggagcag 2461 tggctccaac cgtagcggca gcgatcggag gaaggagaag gacccgaagg ccggggactc 2521 caagtccggg ggcagcggca gcgaatcgga ccacaccaca cgcagcagcc tgcgggggcc 2581 gcgggagcgg gcgcccagcg agcgctcagg tccggcggcc agcgagcaca gccaccgcag 2641 ccaccattcc ctggccagca gccttcgcag ccaccacaca cacccgagct acggtcctcc 2701 cggagtgccc cctctctacg gcccccccat gctgatgatg cccccgccgc ccgcggccat 2761 ggggccccca ggagcccctc cgggccgcga cctggcctca gtgcccccgg aactgaccgc 2821 cagcagacag tccttccgca tggccatggg aaaccccagt gagttctttg tggatgtgat 2881 gtgagcggcc gctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc 2941 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag 3001 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta 3061 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag 3121 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca 3181 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg 3241 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg 3301 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg 3361 gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt 3421 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt 3481 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta 3541 tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa 3601 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg 3661 gtgtggaaag tccccaggct ccccaggcag gcagaagtat gcaaagcatg catctcaatt 3721 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 3781 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 3841 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 3901 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag 3961 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc 4021 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag 4081 gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc 4141 cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga 4201 cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt 4261 ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga 4321 gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat 4381 cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca 4441 cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt 4501 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg 4561 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg 4621 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 4681 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc 4741 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 4801 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 4861 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 4921 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 4981 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 5041 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 5101 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 5161 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 5221 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 5281 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 5341 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 5401 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 5461 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 5521 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 5581 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 5641 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 5701 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 5761 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 5821 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 5881 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 5941 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 6001 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 6061 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 6121 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 6181 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 6241 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 6301 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 6361 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 6421 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 6481 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 6541 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 6601 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 6661 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 6721 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 6781 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 6841 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 6901 ccgaaaagtg ccacctgacg tc //