LOCUS Exported 6480 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expression of Leishmania donovani ectodomain in mammalian cells. ACCESSION . VERSION . KEYWORDS D15_Ld_061240 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6480) TITLE Leishmania ectodomain expression plasmids REFERENCE 2 (bases 1 to 6480) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6480 /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 1258..1275 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 1327..1382 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(1381..1400) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(3618..3636) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3704..3808 /gene="bla" /label=AmpR promoter CDS 3809..4669 /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(4027..4046) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4840..5428 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5329..5348 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5582..5599 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5716..5737 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5752..5782 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5790..5806 /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 5795..5817 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5814..5830 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5814..5830 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" enhancer 6101..6480 /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 gccaccgccg aagcctctag agtggtggaa 841 ctgacccggg acaccttcga cgcctacgtg ggcggacccg acacctacac ctttgtgctg 901 tactgcgtga cctggtcccg gcagtgcaag accgccatcc agctgtggga ccggctgagc 961 atcagccaga gccagaaaga actgcgggac gtgttcaccg ccgcctatgt ggacggcgac 1021 aagtaccccg acctcgtgaa gcggatgaac gtgaagggct tccccaccat gctgttctac 1081 acccccctgc acaccgaggg cctcgagtac aacggcccca gagagagcta cctgctggac 1141 agcttcgtgt tccagttctc tggcgcgccc ggcagcctgc accacatcct ggacgcccag 1201 aagatgctgt ggaaccacag ggacaggaac ctgcctcctc tggcccctct gggcccccac 1261 caccatcacc accattgagt ctagacacgt gattaattaa ggatcccccg acctcgacct 1321 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct 1381 cactcggaag gacatatggg agggcaaatc atttggtcga gatccctcgg agatctctag 1441 ctagaggatc gatccccgcc ccggacgaac taaacctgac tacgacatct ctgccccttc 1501 ttcgcggggc agtgcatgta atcccttcag ttggttggta caacttgcca actgggccct 1561 gttccacatg tgacacgggg ggggaccaaa cacaaagggg ttctctgact gtagttgaca 1621 tccttataaa tggatgtgca catttgccaa cactgagtgg ctttcatcct ggagcagact 1681 ttgcagtctg tggactgcaa cacaacattg cctttatgtg taactcttgg ctgaagctct 1741 tacaccaatg ctgggggaca tgtacctccc aggggcccag gaagactacg ggaggctaca 1801 ccaacgtcaa tcagaggggc ctgtgtagct accgataagc ggaccctcaa gagggcatta 1861 gcaatagtgt ttataaggcc cccttgttaa ccctaaacgg gtagcatatg cttcccgggt 1921 agtagtatat actatccaga ctaaccctaa ttcaatagca tatgttaccc aacgggaagc 1981 atatgctatc gaattagggt tagtaaaagg gtcctaagga acagcgatat ctcccacccc 2041 atgagctgtc acggttttat ttacatgggg tcaggattcc acgagggtag tgaaccattt 2101 tagtcacaag ggcagtggct gaagatcaag gagcgggcag tgaactctcc tgaatcttcg 2161 cctgcttctt cattctcctt cgtttagcta atagaataac tgctgagttg tgaacagtaa 2221 ggtgtatgtg aggtgctcga aaacaaggtt tcaggtgacg cccccagaat aaaatttgga 2281 cggggggttc agtggtggca ttgtgctatg acaccaatat aaccctcaca aaccccttgg 2341 gcaataaata ctagtgtagg aatgaaacat tctgaatatc tttaacaata gaaatccatg 2401 gggtggggac aagccgtaaa gactggatgt ccatctcaca cgaatttatg gctatgggca 2461 acacataatc ctagtgcaat atgatactgg ggttattaag atgtgtccca ggcagggacc 2521 aagacaggtg aaccatgttg ttacactcta tttgtaacaa ggggaaagag agtggacgcc 2581 gacagcagcg gactccactg gttgtctcta acacccccga aaattaaacg gggctccacg 2641 ccaatggggc ccataaacaa agacaagtgg ccactctttt ttttgaaatt gtggagtggg 2701 ggcacgcgtc agcccccaca cgccgccctg cggttttgga ctgtaaaata agggtgtaat 2761 aacttggctg attgtaaccc cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac 2821 taatggcacc ccggggaata cctgcataag taggtgggcg ggccaagata ggggcgcgat 2881 tgctgcgatc tggaggacaa attacacaca cttgcgcctg agcgccaagc acagggttgt 2941 tggtcctcat attcacgagg tcgctgagag cacggtgggc taatgttgcc atgggtagca 3001 tatactaccc aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatagg 3061 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg 3121 ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg 3181 ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg 3241 ctatcctaat agagattagg gtagtatatg ctatcctaat ttatatctgg gtagcatata 3301 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag catatgctat 3361 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat 3421 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat 3481 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat 3541 cctaatctgt atccgggtag catatgctat cctcatgata agctgtcaaa catgagaatt 3601 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat 3661 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg 3721 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 3781 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 3841 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 3901 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 3961 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 4021 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 4081 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 4141 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 4201 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 4261 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 4321 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact 4381 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 4441 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 4501 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 4561 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 4621 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 4681 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 4741 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 4801 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 4861 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 4921 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 4981 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 5041 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 5101 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 5161 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 5221 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 5281 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 5341 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 5401 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 5461 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 5521 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 5581 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc 5641 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag 5701 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt 5761 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa 5821 cagctatgac catgattacg ccaagctcta gctagaggtc gaccaattct catgtttgac 5881 agcttatcat cgcagatccg ggcaacgttg ttgccattgc tgcaggcgca gaactggtag 5941 gtatggaaga tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat 6001 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg 6061 tacatttata ttggctcatg tccaatatga ccgccatgtt gacattgatt attgactagt 6121 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt 6181 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg 6241 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg 6301 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt 6361 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg 6421 accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg //