LOCUS Exported 5134 bp ds-DNA circular SYN 18-DEC-2018 DEFINITION Episomal vector pSpiro-PAC-IFT46-3xHA-C, complete sequence. ACCESSION JQ768101 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5134) AUTHORS Jerlstrom-Hultqvist J, Einarsson E, Svard SG. TITLE Stable Transfection of the Diplomonad Parasite Spironucleus salmonicida JOURNAL Eukaryotic Cell 11 (11), 1353-1361 (2012) PUBMED 22983987 REFERENCE 2 (bases 1 to 5134) AUTHORS Jerlstroem-Hultqvist JSE., Einarsson EM, Svaerd SG. TITLE Direct Submission JOURNAL Submitted (09-MAR-2012) Department of Cell and Molecular Biology, Uppsala University, BMC, Box 596, Uppsala 751 24, Sweden REFERENCE 3 (bases 1 to 5134) AUTHORS . TITLE Direct Submission COMMENT ##Assembly-Data-START## Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END## FEATURES Location/Qualifiers source 1..5134 /organism="Episomal vector pSpiro-PAC-IFT46-3xHA-C" /mol_type="other DNA" /db_xref="taxon:1233395" promoter 24..128 /gene="bla" /label=AmpR promoter CDS 129..989 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" CDS 327..989 /codon_start=1 /transl_except="(pos: 987 .. 989, aa: TERM)" /product="beta-lactamase" /label=beta-lactamase /protein_id="AFS33193.1" /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW" rep_origin 1160..1748 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 2036..2057 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2072..2102 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2110..2126 /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 2134..2150 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 2171..2189 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" 3'UTR complement(2214..2423) /note="ornithine carbamoyltransferase from Spironucleus salmonicida" CDS complement(2421..3029) /codon_start=1 /product="puromycin N-acetyltransferase" /label=puromycin N-acetyltransferase /protein_id="AFS33194.1" /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET SAPRNLPFYERLGFTVTADVECPKDRATWCMTRKPGALE" CDS complement(2430..3029) /codon_start=1 /gene="pac from Streptomyces" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET SAPRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA" 5'UTR complement(3030..3191) /note="ornithine carbamoyltransferase from Spironucleus salmonicida" 5'UTR 3250..3501 /note="intraflagellar transport 46 homolog from Spironucleus salmonicida" CDS 3502..4419 /codon_start=1 /product="intraflagellar transport 46-3xHA" /label=intraflagellar transport 46-3xHA /protein_id="AFS33195.1" /translation="MEMVLILILKWGKSEDILLYITKHKY*IFHFKFSMADNHEDDTLT DEPSDNFATLTNDRLLELFRTFVPTELMLETLFVPFLPDY*PAIGDVDPFIKVGRPDGK LEPLGLIVLDEPCNNQSEPTILESMLEAKTKPIARSKDKTTIKSNKKDKKSQDAKIKAI DYDKEIIHDWVDKVEKIH*T*GSAEYNYSVPVPSATILELWDTEMESIIKTIDLPKPSI ELSIE*YAKVCCLILDIPVGPNAIESLHMFFTNYRDVLDETEVIK*Q*GY*RPPYPYDV PDYAYPYDVPDYAYPYDVPDYAGP" CDS 4330..4410 /codon_start=1 /product="three tandem HA epitope tags" /label=3xHA /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA" 3'UTR 4417..4476 /note="ribosomal protein S15A from Spironucleus salmonicida" promoter complement(4491..4509) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4519..4535) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 4677..5132 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" ORIGIN 1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt 61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa 121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt 181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt 241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt 301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg 361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga 421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa 481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga 541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa 601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca 661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta 721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac 781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc 841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag 901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga 961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt 1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata 1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag 1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc 1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc 1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa 1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg 1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg 1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg 1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat 1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg 2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt 2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg 2161 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct gggtaccact agtgttgtat 2221 ggaatattta tatataaatg cattattaat aatgatatag tatcttaata tacattaata 2281 atttgacaat tccaaacaaa aaagtaacat tattacaagt aataaagtat atattcatta 2341 caaatatatt tcagataagt agtaactgtc tattcattat acaaatgagt aataaaagat 2401 aattaatgaa atttagctga tcactcgagg gcaccgggct tgcgggtcat gcaccaggtc 2461 gcgcggtcct tcgggcactc gacgtcggcg gtgacggtga agccgagccg ctcgtagaag 2521 gggaggttgc ggggcgcgga ggtctccagg aaggcgggca ccccggcgcg ctcggccgcc 2581 tccactccgg ggagcacgac ggcgctgccc agacccttgc cctggtggtc gggcgagacg 2641 ccgacggtgg ccaggaacca cgcgggctcc ttgggccggt gcggcgccag gaggccttcc 2701 atctgttgct gcgcggccag ccgggaaccg ctcaactcgg ccatgcgcgg gccgatctcg 2761 gcgaacaccg cccccgcttc gacgctctcc ggcgtggtcc agaccgccac cgcggcgccg 2821 tcgtccgcga cccacacctt gccgatgtcg agcccgacgc gcgtgaggaa gagttcttgc 2881 agctcggtga cccgctcgat gtggcggtcc gggtcgacgg tgtggcgcgt ggcggggtag 2941 tcggcgaacg cggcggcgag ggtgcgtacg gcccggggga cgtcgtcgcg ggtggcgagg 3001 cgcaccgtgg gcttgtactc ggtgcccatg gtgggaagtg gtgaccaaat ctagaaaaaa 3061 gatttaagaa tattatttgt taaattatta atctctatta tttccccctc cattatgttc 3121 aaaaatatat ataaatatac cataattatt ctacttattt tgtgattttc ttaaatatgt 3181 gagtatgcct ccatatgaag cttgcttaat taagcgctag cgatatcgga tcctgcagtc 3241 tagacgcgtc gttctcgttc ggataaaatg ctgttcaaaa gtaagaggag gtgaagcctg 3301 ctgagatgac ttctttcaag ttcggaaatg ctcaggtgac tgccgaaaaa cccatgtttg 3361 gggcagtttc atagcaagaa tagcagaaac cagcagaata ggtataattt aagtttggaa 3421 acgcgcaggt aactgctgat aagcctatgt tcggagcacc tgtcaaaccg gcagagtaga 3481 gcgcacctgc ttttggcgat aatggaaatg gttttaattt taattttaaa atggggtaaa 3541 agtgaagata ttttattata tattactaaa cataaatact aaatttttca cttcaaattt 3601 tcaatggcgg ataaccacga agacgatacc ttaacagatg agccctcaga caactttgca 3661 actcttacca atgataggct tctagaatta ttcagaactt ttgttcccac agagctcatg 3721 cttgaaactc tttttgtacc ttttttacca gattattaac cggctattgg tgatgtagat 3781 ccattcataa aggttggccg tccagatggt aaattagaac ctttaggctt aattgtttta 3841 gatgaaccat gtaataacca atctgagcct actattttag aatctatgct agaagcaaaa 3901 acgaaaccga ttgccaggtc taaagataaa acaactatta aaagtaataa aaaagataaa 3961 aaatcacagg atgctaagat taaagctatt gattatgaca aggaaattat acatgattgg 4021 gtagataaag ttgaaaaaat tcattaaaca tagggttctg cagaatataa ttattcagta 4081 ccagttccaa gtgctacaat attagaattg tgggatactg aaatggagtc tattatcaaa 4141 accatagatt tgccaaaacc aagtatcgaa ctttccatcg aataatatgc taaagtatgt 4201 tgccttattt tagatattcc agtaggacct aatgctattg aaagtttaca tatgttcttt 4261 actaattata gggatgtatt agatgaaact gaagttataa aataacaata aggatattag 4321 cggccgccct atccctacga cgtgccagat tacgcatatc cgtacgatgt tcctgactat 4381 gcttacccct atgatgtccc tgactacgcc gggccctgat taaatttgat ttttttaaat 4441 aaactattat tccaaaattt tgatttatat tatttggagc tccaattcgc cctatagtga 4501 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg 4561 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga 4621 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc 4681 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 4741 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 4801 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 4861 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc 4921 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 4981 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg 5041 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 5101 gaattttaac aaaatattaa cgcttacaat ttag //