LOCUS Exported 6978 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expression of Leishmania donovani ectodomain in mammalian cells. ACCESSION . VERSION . KEYWORDS D29_Ld_251870 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6978) TITLE Leishmania ectodomain expression plasmids REFERENCE 2 (bases 1 to 6978) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6978 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 581..605 /label=LNCX /note="Human CMV promoter, forward primer" primer_bind 1086..1105 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 1128..1137 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2136..2153 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 2205..2260 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(2259..2278) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4496..4514) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4582..4686 /gene="bla" /label=AmpR promoter CDS 4687..5547 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(4905..4924) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5718..6306 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6207..6226 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6460..6477 /label=L4440 /note="L4440 vector, forward primer" protein_bind 6594..6615 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6630..6660 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6668..6684 /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)." primer_bind 6673..6695 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 6692..6708 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 6692..6708 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc 601 agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc tcgcggttga 661 ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg gcctccgaac 721 ggtactccgc caccgaggga cctgagcgag tccgcatcga ccggatcgga aaacctctcg 781 agaaaggcgt ctaaccagtc acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc 841 gggtggcggt cggggttgtt tctggcggag gtgctgctga tgatgtaatt aaagtaggcg 901 gtcttgagac ggcggatggt cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg 961 tgagtactcc ctctcaaaag cgggcattac ttctgcgcta agattgtcag tttccaaaaa 1021 cgaggaggat ttgatattca cctggcccga tctggccata cacttgagtg acaatgacat 1081 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caagtttgcc accatggagt 1141 ttcagaccca ggtactcatg tccctgctgc tctgcatgtc tggtgcggcc gccaccgccc 1201 tgtactacta cagcgtgtac gactacgaga agcacatgag catggtgcag cggaagtaca 1261 gcggctacga ccccctgacc gactgcgcca ccccttttgg ccagctgctg ggcgtggccg 1321 acgatgtgcc cgcctacagc aactgcaaca ccaagttcag cagcacctac atcaactacg 1381 tgtccctgat ggaccccatg gacaacggca gacggggcga ccccagcgag acacggatcg 1441 tgatgaccgc ctaccggtac accgccttcg actactgcat gcggtggctc gtgtggaacc 1501 ggggcgtgat gccccggctg gtggaaaaca ccaaccagct ctggaaaacc gtggactact 1561 tcaaccccgc caagcccgag caggggtggt ccgccgagta catcaccaac tacgaagagg 1621 tcacaggcat ggaagagaga aagttcaacg cccccagacg cgccgacgcc atcgtgtacc 1681 ggatggacaa gaacaccatc cctgccggcc acattgccgt ggtggtcaag gtggaagatg 1741 acgtggaagc cgctggcggc cctgagaagc tgaaagaact gaagaagatg cggctgcacc 1801 ccaggcgggt gtacgtggca gagcagaatt ggaagaacca gccctggggc ggacacaact 1861 acagccgggt gctgcagttc aagtggcgcg ctgtggccga gaaggcctac gagggctgct 1921 acgtggaccc cgacggcctg ggcatcatcg gcgtcgtcag agtgggcaag gccatgcccc 1981 tgagagccgc cccagacccc taccaggaag ccctggacat ggataacgac ggcgacctgg 2041 gcgcgcccgg cagcctgcac cacatcctgg acgcccagaa gatgctgtgg aaccacaggg 2101 acaggaacct gcctcctctg gcccctctgg gcccccacca ccatcaccac cattgagtct 2161 agacacgtga ttaattaagg atcccccgac ctcgacctct ggctaataaa ggaaatttat 2221 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag 2281 ggcaaatcat ttggtcgaga tccctcggag atctctagct agaggatcga tccccgcccc 2341 ggacgaacta aacctgacta cgacatctct gccccttctt cgcggggcag tgcatgtaat 2401 cccttcagtt ggttggtaca acttgccaac tgggccctgt tccacatgtg acacgggggg 2461 ggaccaaaca caaaggggtt ctctgactgt agttgacatc cttataaatg gatgtgcaca 2521 tttgccaaca ctgagtggct ttcatcctgg agcagacttt gcagtctgtg gactgcaaca 2581 caacattgcc tttatgtgta actcttggct gaagctctta caccaatgct gggggacatg 2641 tacctcccag gggcccagga agactacggg aggctacacc aacgtcaatc agaggggcct 2701 gtgtagctac cgataagcgg accctcaaga gggcattagc aatagtgttt ataaggcccc 2761 cttgttaacc ctaaacgggt agcatatgct tcccgggtag tagtatatac tatccagact 2821 aaccctaatt caatagcata tgttacccaa cgggaagcat atgctatcga attagggtta 2881 gtaaaagggt cctaaggaac agcgatatct cccaccccat gagctgtcac ggttttattt 2941 acatggggtc aggattccac gagggtagtg aaccatttta gtcacaaggg cagtggctga 3001 agatcaagga gcgggcagtg aactctcctg aatcttcgcc tgcttcttca ttctccttcg 3061 tttagctaat agaataactg ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa 3121 acaaggtttc aggtgacgcc cccagaataa aatttggacg gggggttcag tggtggcatt 3181 gtgctatgac accaatataa ccctcacaaa ccccttgggc aataaatact agtgtaggaa 3241 tgaaacattc tgaatatctt taacaataga aatccatggg gtggggacaa gccgtaaaga 3301 ctggatgtcc atctcacacg aatttatggc tatgggcaac acataatcct agtgcaatat 3361 gatactgggg ttattaagat gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt 3421 acactctatt tgtaacaagg ggaaagagag tggacgccga cagcagcgga ctccactggt 3481 tgtctctaac acccccgaaa attaaacggg gctccacgcc aatggggccc ataaacaaag 3541 acaagtggcc actctttttt ttgaaattgt ggagtggggg cacgcgtcag cccccacacg 3601 ccgccctgcg gttttggact gtaaaataag ggtgtaataa cttggctgat tgtaaccccg 3661 ctaaccactg cggtcaaacc acttgcccac aaaaccacta atggcacccc ggggaatacc 3721 tgcataagta ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat 3781 tacacacact tgcgcctgag cgccaagcac agggttgttg gtcctcatat tcacgaggtc 3841 gctgagagca cggtgggcta atgttgccat gggtagcata tactacccaa atatctggat 3901 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt 3961 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt 4021 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt 4081 agtatatgct atcctaatct gtatccgggt agcatatgct atcctaatag agattagggt 4141 agtatatgct atcctaattt atatctgggt agcatatact acccaaatat ctggatagca 4201 tatgctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagca 4261 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta 4321 tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca 4381 tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca 4441 tatgctatcc tcatgataag ctgtcaaaca tgagaatttt cttgaagacg aaagggcctc 4501 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt 4561 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca 4621 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg 4681 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc 4741 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg 4801 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt 4861 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta 4921 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat 4981 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga 5041 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca 5101 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact 5161 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc 5221 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact 5281 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt 5341 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt 5401 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt 5461 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata 5521 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag 5581 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat 5641 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa 5701 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 5761 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 5821 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg 5881 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 5941 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 6001 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 6061 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 6121 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 6181 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 6241 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 6301 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 6361 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag 6421 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa 6481 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc 6541 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg 6601 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg 6661 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc 6721 aagctctagc tagaggtcga ccaattctca tgtttgacag cttatcatcg cagatccggg 6781 caacgttgtt gccattgctg caggcgcaga actggtaggt atggaagatc tatacattga 6841 atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct 6901 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc 6961 caatatgacc gccatgtt //