LOCUS Exported 7632 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expression of Leishmania donovani ectodomain in mammalian cells. ACCESSION . VERSION . KEYWORDS D31_Ld_261020 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7632) TITLE Leishmania ectodomain expression plasmids REFERENCE 2 (bases 1 to 7632) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7632 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..204 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 155..175 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 201..225 /label=LNCX /note="Human CMV promoter, forward primer" primer_bind 706..725 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 748..757 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2410..2427 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 2479..2534 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(2533..2552) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4770..4788) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4856..4960 /gene="bla" /label=AmpR promoter CDS 4961..5821 /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(5179..5198) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5992..6580 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6481..6500 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6734..6751 /label=L4440 /note="L4440 vector, forward primer" protein_bind 6868..6889 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6904..6934 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6942..6958 /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 6947..6969 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 6966..6982 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 6966..6982 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" enhancer 7253..7632 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt 61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac 121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg 181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcctcac tctcttccgc 241 atcgctgtct gcgagggcca gctgttgggc tcgcggttga ggacaaactc ttcgcggtct 301 ttccagtact cttggatcgg aaacccgtcg gcctccgaac ggtactccgc caccgaggga 361 cctgagcgag tccgcatcga ccggatcgga aaacctctcg agaaaggcgt ctaaccagtc 421 acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc gggtggcggt cggggttgtt 481 tctggcggag gtgctgctga tgatgtaatt aaagtaggcg gtcttgagac ggcggatggt 541 cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg tgagtactcc ctctcaaaag 601 cgggcattac ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca 661 cctggcccga tctggccata cacttgagtg acaatgacat ccactttgcc tttctctcca 721 caggtgtcca ctcccaggtc caagtttgcc accatggagt ttcagaccca ggtactcatg 781 tccctgctgc tctgcatgtc tggtgcggcc gccaccggac ctgtgctgga agtgggagtg 841 gaccgggcca tcaccttcaa cagacagctg ggcgacggcg acgagctgct gttcggaagc 901 agagccgtgc tgctgcgggg cgagatcgtg cggtatacag ccggcgcttc cgccagctac 961 cacggctaca ccgtgtacgt gcagtgcagc gagcacctga ccaacttcgc cctgcagcag 1021 ctggccgctc ccgagctgtc taaggccgcc aagggcctgg tggtggaact gtgcgacacc 1081 gacaccacca acgaggacgt gctgagcttc ttcttctcca acgccgctgc cagcatcccc 1141 gtgtactttc tgcccaacgg caccgccagc aaagagctga gcagactgct gagcgtggcc 1201 gcccagagaa gcgccaccga gagggtggtg ctgtccgtgg ccaagaccag gcggctgagc 1261 gagctggtgg aaaacagcac cctgccctcc gccaccatcg agagccacta cgtgcacaag 1321 cccaagcaga ccagaagcaa agccgccagc gctgccgccg ctcctcaggt gctggtcacc 1381 gcccacttcg actccctggg cgtgtcccca gccagcagaa cctctggcgg aaccagcgga 1441 gccgtggtgg ccatggaact gtggcggaga ctgaccagca ccccctacgc cagacaggaa 1501 agcgtcgccc cttacgaagt gaccgtcctg ctgggcagca ccagccggtt caactacgcc 1561 ggcaccacaa gctggctgag ccagcacacc gatagagagc tggaccagtt ccgggtggtc 1621 ctgtgcctgg acgaactgct gccccccaga gagacaagcg agaacgcccc cgacctgtat 1681 ctgcatgtgc aggatgtgct gatgaagcgg cagcacggcc agcaggtcgt ggaacaggtg 1741 gaagccgccg caaagctgct gggaatcagc ctgaaggtgg tgtccgccaa gacaaactac 1801 cagcactacg acctggaatt cgagcacgag gccttcgcca gccggcagat gaccgccatg 1861 accctgagca cccaccgggt gcaccataac gaccagctgt tccgggacct gcggaggcct 1921 cctgccacag ccgctgatgc cgctatgctg gccaagcggg ccgacctggt cgaggccatt 1981 gtgcggattg tggccgaggc tgcccctagc gccgagggcg tgacaacagt gtggcctggc 2041 gccgctagct acatccaggg catgctgcac tacgcctccg acagccagag atcccccgtg 2101 gcccagaacg gcgctggcct gagacagtac gccgccacag tggaacacca catgagagcc 2161 caggccgctg ctgctcagag agccgccttt accagcgcca gctctgtggc cacctaccag 2221 gaactgcgga cccccggcat caccctgttc ggcccatacg aagagacaat gcaggtgttc 2281 atggccaagt cctacctgtt cgagtgcgcc gtgggcgcgc ccggcagcct gcaccacatc 2341 ctggacgccc agaagatgct gtggaaccac agggacagga acctgcctcc tctggcccct 2401 ctgggccccc accaccatca ccaccattga gtctagacac gtgattaatt aaggatcccc 2461 cgacctcgac ctctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt 2521 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttggtc gagatccctc 2581 ggagatctct agctagagga tcgatccccg ccccggacga actaaacctg actacgacat 2641 ctctgcccct tcttcgcggg gcagtgcatg taatcccttc agttggttgg tacaacttgc 2701 caactgggcc ctgttccaca tgtgacacgg ggggggacca aacacaaagg ggttctctga 2761 ctgtagttga catccttata aatggatgtg cacatttgcc aacactgagt ggctttcatc 2821 ctggagcaga ctttgcagtc tgtggactgc aacacaacat tgcctttatg tgtaactctt 2881 ggctgaagct cttacaccaa tgctggggga catgtacctc ccaggggccc aggaagacta 2941 cgggaggcta caccaacgtc aatcagaggg gcctgtgtag ctaccgataa gcggaccctc 3001 aagagggcat tagcaatagt gtttataagg cccccttgtt aaccctaaac gggtagcata 3061 tgcttcccgg gtagtagtat atactatcca gactaaccct aattcaatag catatgttac 3121 ccaacgggaa gcatatgcta tcgaattagg gttagtaaaa gggtcctaag gaacagcgat 3181 atctcccacc ccatgagctg tcacggtttt atttacatgg ggtcaggatt ccacgagggt 3241 agtgaaccat tttagtcaca agggcagtgg ctgaagatca aggagcgggc agtgaactct 3301 cctgaatctt cgcctgcttc ttcattctcc ttcgtttagc taatagaata actgctgagt 3361 tgtgaacagt aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga cgcccccaga 3421 ataaaatttg gacggggggt tcagtggtgg cattgtgcta tgacaccaat ataaccctca 3481 caaacccctt gggcaataaa tactagtgta ggaatgaaac attctgaata tctttaacaa 3541 tagaaatcca tggggtgggg acaagccgta aagactggat gtccatctca cacgaattta 3601 tggctatggg caacacataa tcctagtgca atatgatact ggggttatta agatgtgtcc 3661 caggcaggga ccaagacagg tgaaccatgt tgttacactc tatttgtaac aaggggaaag 3721 agagtggacg ccgacagcag cggactccac tggttgtctc taacaccccc gaaaattaaa 3781 cggggctcca cgccaatggg gcccataaac aaagacaagt ggccactctt ttttttgaaa 3841 ttgtggagtg ggggcacgcg tcagccccca cacgccgccc tgcggttttg gactgtaaaa 3901 taagggtgta ataacttggc tgattgtaac cccgctaacc actgcggtca aaccacttgc 3961 ccacaaaacc actaatggca ccccggggaa tacctgcata agtaggtggg cgggccaaga 4021 taggggcgcg attgctgcga tctggaggac aaattacaca cacttgcgcc tgagcgccaa 4081 gcacagggtt gttggtcctc atattcacga ggtcgctgag agcacggtgg gctaatgttg 4141 ccatgggtag catatactac ccaaatatct ggatagcata tgctatccta atctatatct 4201 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct 4261 gggtagtata tgctatccta atttatatct gggtagcata ggctatccta atctatatct 4321 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc 4381 gggtagcata tgctatccta atagagatta gggtagtata tgctatccta atttatatct 4441 gggtagcata tactacccaa atatctggat agcatatgct atcctaatct atatctgggt 4501 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt 4561 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt 4621 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt 4681 agtatatgct atcctaatct gtatccgggt agcatatgct atcctcatga taagctgtca 4741 aacatgagaa ttttcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa 4801 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg 4861 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata 4921 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg 4981 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac 5041 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact 5101 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat 5161 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga 5221 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac 5281 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat 5341 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac 5401 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct 5461 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac 5521 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga 5581 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg 5641 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact 5701 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac 5761 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta 5821 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt 5881 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga 5941 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc 6001 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt 6061 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc 6121 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc 6181 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg 6241 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg 6301 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga 6361 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc 6421 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg 6481 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg 6541 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt 6601 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc 6661 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg 6721 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc 6781 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg 6841 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca 6901 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt 6961 tcacacagga aacagctatg accatgatta cgccaagctc tagctagagg tcgaccaatt 7021 ctcatgtttg acagcttatc atcgcagatc cgggcaacgt tgttgccatt gctgcaggcg 7081 cagaactggt aggtatggaa gatctataca ttgaatcaat attggcaatt agccatatta 7141 gtcattggtt atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta 7201 tatcataata tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga 7261 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg 7321 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc 7381 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat 7441 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat 7501 catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat 7561 gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc 7621 gctattacca tg //