LOCUS Exported 6555 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expression of Leishmania donovani ectodomain in mammalian cells. ACCESSION . VERSION . KEYWORDS D12_Ld_040370 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6555) TITLE Leishmania ectodomain expression plasmids REFERENCE 2 (bases 1 to 6555) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6555 /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 1333..1350 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 1402..1457 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(1456..1475) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(3693..3711) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3779..3883 /gene="bla" /label=AmpR promoter CDS 3884..4744 /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(4102..4121) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4915..5503 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5404..5423 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5657..5674 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5791..5812 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5827..5857 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5865..5881 /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 5870..5892 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5889..5905 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5889..5905 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" enhancer 6176..6555 /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 gccaccgata tgctgctggc ccccagcgac 841 agcatcccct tcaccacctt cggccagctg cagcaggtct gcgccgccac acaggccgga 901 agcacccctg tggccatgta caccgccacc atgaccgccc tggtggccaa cctgatgagc 961 gagaagaaag tgacccacgc ctacgtgtcc gcctacctga acgccgaccg gcggtggact 1021 tggtttgtgt acgagggcga cgaagtgatc gagatgctga tgaacagcag ccagtgggcc 1081 accggcaacc ccacaggcgc ccacaccacc taccggcacg tggtgtacag caccaccgac 1141 aagggcctga tcagcgccac aggctacgac aactaccccg tcgtgtgcaa atacgagaac 1201 cagaccgact acagcccccc tggcaagcgg ttccctggcg cgcccggcag cctgcaccac 1261 atcctggacg cccagaagat gctgtggaac cacagggaca ggaacctgcc tcctctggcc 1321 cctctgggcc cccaccacca tcaccaccat tgagtctaga cacgtgatta attaaggatc 1381 ccccgacctc gacctctggc taataaagga aatttatttt cattgcaata gtgtgttgga 1441 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcatttg gtcgagatcc 1501 ctcggagatc tctagctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga 1561 catctctgcc ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact 1621 tgccaactgg gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc 1681 tgactgtagt tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc 1741 atcctggagc agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact 1801 cttggctgaa gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga 1861 ctacgggagg ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc 1921 ctcaagaggg cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc 1981 atatgcttcc cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt 2041 tacccaacgg gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc 2101 gatatctccc accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag 2161 ggtagtgaac cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac 2221 tctcctgaat cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg 2281 agttgtgaac agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc 2341 agaataaaat ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc 2401 tcacaaaccc cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa 2461 caatagaaat ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat 2521 ttatggctat gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg 2581 tcccaggcag ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga 2641 aagagagtgg acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt 2701 aaacggggct ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg 2761 aaattgtgga gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta 2821 aaataagggt gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact 2881 tgcccacaaa accactaatg gcaccccggg gaatacctgc ataagtaggt gggcgggcca 2941 agataggggc gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc 3001 caagcacagg gttgttggtc ctcatattca cgaggtcgct gagagcacgg tgggctaatg 3061 ttgccatggg tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata 3121 tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata 3181 tctgggtagt atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata 3241 tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta 3301 tccgggtagc atatgctatc ctaatagaga ttagggtagt atatgctatc ctaatttata 3361 tctgggtagc atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg 3421 ggtagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg 3481 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg 3541 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg 3601 ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctca tgataagctg 3661 tcaaacatga gaattttctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt 3721 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg 3781 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca 3841 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt 3901 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga 3961 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga 4021 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat 4081 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca 4141 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt 4201 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac 4261 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct 4321 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga 4381 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac 4441 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat 4501 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg 4561 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc 4621 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc 4681 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg 4741 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta 4801 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg 4861 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga 4921 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt 4981 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag 5041 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa 5101 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag 5161 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca 5221 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac 5281 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa 5341 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc 5401 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg 5461 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc 5521 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc 5581 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag 5641 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa 5701 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga 5761 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc 5821 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca 5881 atttcacaca ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacca 5941 attctcatgt ttgacagctt atcatcgcag atccgggcaa cgttgttgcc attgctgcag 6001 gcgcagaact ggtaggtatg gaagatctat acattgaatc aatattggca attagccata 6061 ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat 6121 ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgttgacat 6181 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat 6241 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac 6301 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc 6361 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg 6421 tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat 6481 tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc 6541 atcgctatta ccatg //