LOCUS Exported 6172 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS XE252 pCDNA3.1(zeo)-hDsh3-333-716 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6172) TITLE Xenopus Expression Plasmids REFERENCE 2 (bases 1 to 6172) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6172 /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" primer_bind complement(2179..2196) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2185..2409 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2455..2883 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2542..2561) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2752..2773 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2892..2912) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2897..3227 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3078..3213 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3140..3159 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 3275..3322 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 3341..3715 /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 3845..3966 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3882..3901) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3936..3955 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4015..4031) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4015..4031) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4028..4050) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4039..4055 /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(4063..4093) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4108..4129 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4246..4263) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4417..5005) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4497..4516) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5176..6036) /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 5799..5818 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6037..6141) /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 tagccaagtg ctgggaccca agtccacgtg gttgcttcac 1021 attgcccagg agcgagccca tccggcccat tgaccctgcg gcctgggtct cccacactgc 1081 agccatgacc ggcaccttcc ctgcatacgg catgagcccc tccctgagca ccatcacctc 1141 caccagctcc tccatcacca gttccatccc tgacacagag cgcctagacg acttccactt 1201 gtccatccac agtgacatgg ctgccatcgt aaaagccatg gcctcccctg aatcagggtt 1261 ggaggtccgt gaccgcatgt ggctcaagat taccatccct aatgctttca tcggctcaga 1321 tgtggtggac tggctgtacc acaatgtgga aggcttcacg gaccggaggg aggcccgcaa 1381 gtatgccagc aacctgctga aagctggctt catccgccat accgtcaaca agatcacctt 1441 ctccgagcag tgctactaca tcttcggtga cctctgcggc aacatggcca acctgtctct 1501 ccacgatcac gatggctcca gtggcgcctc tgaccaggac acactggccc ctttgccgca 1561 cccgggggcc gccccttggc ccatggcttt cccgtaccag tacccgccac ccccgcaccc 1621 atacaacccg cacccgggct tcccggagct gggctacagc tacggcgggg gcagcgccag 1681 cagtcagcac agcgaaggca gtcggagcag tggctccaac cgtagcggca gcgatcggag 1741 gaaggagaag gacccgaagg ccggggactc caagtccggg ggcagcggca gcgaatcgga 1801 ccacaccaca cgcagcagcc tgcgggggcc gcgggagcgg gcgcccagcg agcgctcagg 1861 tccggcggcc agcgagcaca gccaccgcag ccaccattcc ctggccagca gccttcgcag 1921 ccaccacaca cacccgagct acggtcctcc cggagtgccc cctctctacg gcccccccat 1981 gctgatgatg cccccgccgc ccgcggccat ggggccccca ggagcccctc cgggccgcga 2041 cctggcctca gtgcccccgg aactgaccgc cagcagacag tccttccgca tggccatggg 2101 aaaccccagt gagttctttg tggatgtgat gtgagcggcc gctcgagtct agagggcccg 2161 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc 2221 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 2281 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 2341 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg 2401 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc 2461 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 2521 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 2581 ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt 2641 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 2701 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 2761 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga 2821 ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 2881 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccaggcag 2941 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 3001 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 3061 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 3121 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat 3181 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag 3241 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta 3301 tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc 3361 cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct 3421 cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac 3481 cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg 3541 ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg 3601 ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc 3661 cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt 3721 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt 3781 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca 3841 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 3901 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 3961 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata 4021 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag 4081 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg 4141 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca 4201 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 4261 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 4321 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 4381 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 4441 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 4501 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 4561 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg 4621 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 4681 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 4741 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 4801 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac 4861 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 4921 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat 4981 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 5041 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 5101 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 5161 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 5221 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg 5281 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga 5341 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt 5401 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 5461 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt 5521 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 5581 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc 5641 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 5701 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 5761 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag 5821 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt 5881 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 5941 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 6001 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg 6061 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa 6121 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc //