LOCUS Exported 7800 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expression of Leishmania donovani ectodomain in mammalian cells. ACCESSION . VERSION . KEYWORDS D3_Ld_030930 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7800) TITLE Leishmania ectodomain expression plasmids REFERENCE 2 (bases 1 to 7800) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7800 /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 2578..2595 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 2647..2702 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(2701..2720) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4938..4956) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5024..5128 /gene="bla" /label=AmpR promoter CDS 5129..5989 /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(5347..5366) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6160..6748 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6649..6668 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6902..6919 /label=L4440 /note="L4440 vector, forward primer" protein_bind 7036..7057 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 7072..7102 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7110..7126 /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 7115..7137 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7134..7150 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7134..7150 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" enhancer 7421..7800 /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 gccaccgctg atgctgtgtc cagccccgtg 841 atcctgccca gaggccggtc caacaccaaa gtgaccctgt acgtgcggac cggcagagtg 901 tccatccccc tggaatggat cgacggcaaa gaggtgttct tcgagtacac cggcagactg 961 tacgaagtgg gcaacagcgg ccccatgctg cctggcccca ccctgaaagt gaaccctggc 1021 ggcagaatcg tgctgaagct ggtcaacgac ctgggcaaag agggcatggc caacatgacc 1081 ggccggatga acaccctgca cggccccaac atcaccaacg tgcacttcca cggcatgcac 1141 agcgacccca agatggacaa ccccttcaag gtggccctgc ccggcgagac actggtgtac 1201 acaatcaacg tgccccggga ccacgagccc ggcctgcact ggtatcacgc ccacagccac 1261 ggggccgtgt actaccacgt gatgggcggc atgttcggcg ccatcgacgt gggcgagggc 1321 gacttcacca agacccccag acaccccttc cacggctggg acagccagat cctgatggtg 1381 cacctgtacc ggctgaacac cagcgagaga tgcgacggca tgcccatgag cagcctggac 1441 gccgccatgt ccagcctgct gcccttcgac ccccggatcg tggaccggaa gggcaacgag 1501 tacgagatgc ccgccgagct gttcctggtc aacggccagc accggcccac cgtcagagtg 1561 aagaggggcc accctatgct gctgcggatc ggctttgccg ccggaagctg ctacatcaac 1621 atgagcctgc ccaagcagtg cgccttccac atgacagcca tcgacggcgc ccagctgccc 1681 cggaccgtgg aagtggtgga aggctggctg tacttcacca ccgccacccg gcgaagcctg 1741 gtggtcgtgt gtgacgaaga gggcacctac cccgtgtccc acaccggcga tcccagcgac 1801 gccatcttct acctgaagtc ccggggcaag caccagaaag gcggcagagc cgtggccttc 1861 cccgtgacca tgcacaagta cagccccggc tacctgcagc tgaagggcca gaacaccttc 1921 tacagagaga tcagcttcag ccagcgggac ctgcccatca ccaagcccta ctacgtgctg 1981 ggccagggcc ccaactgcca gagcctgcag aacagcagca cctgtcacta cgagtacttc 2041 cagggcgaga tcggcagacg gatcgagggc taccacggct tcaccgtgcc tctgcacgcc 2101 gtggtcaccg ccagagtgta cggcgacccc accgacaaga ggccccaccc cctgcacttt 2161 cacgtgaacc acttccagtt tctgagcttc gagcccagac ctggcggcaa ccacgagaac 2221 gagacaatgg ctatgtacgg cgtgcactcc ggcgagttcc gggacaccat ccccatcctg 2281 gacggcgtga ccaccatccg gtggaaggcc gccacctatg tgggcgaggt ggtgtaccac 2341 tgccacgccc tgcagcacga ggaccggggc atgatgatga gctacctggt gtacagcccc 2401 cggtccaaag agggaaaggc cgtccagaga tacgagcaga gccagaccgg cgccttcctg 2461 ctgacctggc ctcacagatc tggcgcgccc ggcagcctgc accacatcct ggacgcccag 2521 aagatgctgt ggaaccacag ggacaggaac ctgcctcctc tggcccctct gggcccccac 2581 caccatcacc accattgagt ctagacacgt gattaattaa ggatcccccg acctcgacct 2641 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct 2701 cactcggaag gacatatggg agggcaaatc atttggtcga gatccctcgg agatctctag 2761 ctagaggatc gatccccgcc ccggacgaac taaacctgac tacgacatct ctgccccttc 2821 ttcgcggggc agtgcatgta atcccttcag ttggttggta caacttgcca actgggccct 2881 gttccacatg tgacacgggg ggggaccaaa cacaaagggg ttctctgact gtagttgaca 2941 tccttataaa tggatgtgca catttgccaa cactgagtgg ctttcatcct ggagcagact 3001 ttgcagtctg tggactgcaa cacaacattg cctttatgtg taactcttgg ctgaagctct 3061 tacaccaatg ctgggggaca tgtacctccc aggggcccag gaagactacg ggaggctaca 3121 ccaacgtcaa tcagaggggc ctgtgtagct accgataagc ggaccctcaa gagggcatta 3181 gcaatagtgt ttataaggcc cccttgttaa ccctaaacgg gtagcatatg cttcccgggt 3241 agtagtatat actatccaga ctaaccctaa ttcaatagca tatgttaccc aacgggaagc 3301 atatgctatc gaattagggt tagtaaaagg gtcctaagga acagcgatat ctcccacccc 3361 atgagctgtc acggttttat ttacatgggg tcaggattcc acgagggtag tgaaccattt 3421 tagtcacaag ggcagtggct gaagatcaag gagcgggcag tgaactctcc tgaatcttcg 3481 cctgcttctt cattctcctt cgtttagcta atagaataac tgctgagttg tgaacagtaa 3541 ggtgtatgtg aggtgctcga aaacaaggtt tcaggtgacg cccccagaat aaaatttgga 3601 cggggggttc agtggtggca ttgtgctatg acaccaatat aaccctcaca aaccccttgg 3661 gcaataaata ctagtgtagg aatgaaacat tctgaatatc tttaacaata gaaatccatg 3721 gggtggggac aagccgtaaa gactggatgt ccatctcaca cgaatttatg gctatgggca 3781 acacataatc ctagtgcaat atgatactgg ggttattaag atgtgtccca ggcagggacc 3841 aagacaggtg aaccatgttg ttacactcta tttgtaacaa ggggaaagag agtggacgcc 3901 gacagcagcg gactccactg gttgtctcta acacccccga aaattaaacg gggctccacg 3961 ccaatggggc ccataaacaa agacaagtgg ccactctttt ttttgaaatt gtggagtggg 4021 ggcacgcgtc agcccccaca cgccgccctg cggttttgga ctgtaaaata agggtgtaat 4081 aacttggctg attgtaaccc cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac 4141 taatggcacc ccggggaata cctgcataag taggtgggcg ggccaagata ggggcgcgat 4201 tgctgcgatc tggaggacaa attacacaca cttgcgcctg agcgccaagc acagggttgt 4261 tggtcctcat attcacgagg tcgctgagag cacggtgggc taatgttgcc atgggtagca 4321 tatactaccc aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatagg 4381 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg 4441 ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg 4501 ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg 4561 ctatcctaat agagattagg gtagtatatg ctatcctaat ttatatctgg gtagcatata 4621 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag catatgctat 4681 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat 4741 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat 4801 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat 4861 cctaatctgt atccgggtag catatgctat cctcatgata agctgtcaaa catgagaatt 4921 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat 4981 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg 5041 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 5101 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 5161 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 5221 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 5281 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 5341 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 5401 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 5461 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 5521 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 5581 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 5641 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact 5701 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 5761 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 5821 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 5881 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 5941 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 6001 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 6061 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 6121 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 6181 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 6241 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 6301 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 6361 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 6421 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 6481 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 6541 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 6601 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 6661 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 6721 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 6781 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 6841 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 6901 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc 6961 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag 7021 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt 7081 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa 7141 cagctatgac catgattacg ccaagctcta gctagaggtc gaccaattct catgtttgac 7201 agcttatcat cgcagatccg ggcaacgttg ttgccattgc tgcaggcgca gaactggtag 7261 gtatggaaga tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat 7321 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg 7381 tacatttata ttggctcatg tccaatatga ccgccatgtt gacattgatt attgactagt 7441 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt 7501 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg 7561 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg 7621 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt 7681 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg 7741 accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg //