LOCUS Exported 5415 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Homology arms and mTagRFPT-linker sequence for N-terminus tagging of human CETN2. ACCESSION . VERSION . KEYWORDS AICSDP-22:CETN2-mTagRFP-T SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5415) AUTHORS Allen Institute for Cell Science TITLE Allen Institute for Cell Science Plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 5415) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5415 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 56..75 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 309..326 /label=L4440 /note="L4440 vector, forward primer" protein_bind 443..464 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 479..509 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 517..533 /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 522..544 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 541..557 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 541..557 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS complement(1607..2335) /codon_start=1 /product="photostable variant of TagRFP (Shaner et al., 2008)" /label=TagRFP-T /translation="VSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMR IKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV LTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTDM ALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVA RYCDLPSKLGHKLNGMDELYK" primer_bind complement(3356..3373) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3356..3372) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3365..3387) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(3581..3600) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3700..3722 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3760..3778) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3846..3950 /gene="bla" /label=AmpR promoter CDS 3951..4811 /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" primer_bind complement(4169..4188) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4982..155 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 61 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 121 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac 181 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt 241 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga 301 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc 361 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag 421 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt 481 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca 541 caggaaacag ctatgaccat gattacgcca agctcggcgc gccattggga tcaggtcacc 601 ctttcctttt taacaatgta actcgagtct agacttgcgg caagtgtcag gaagcccatt 661 ttggcctcaa agaaacacca ggcaagccag gcactccatc caaggtgacg tttgctacct 721 atcgctatat ctctccctat cccatatcta tttcctaact gatttggaat tctgtgggaa 781 gaggtatctg cttggaagtc ttgagaaaga agcacaaggg cttgttttgg gagcccaaag 841 taacatgatg ggagggcagg cctcctgatg cagttgctac ctggactgct ctgtggtcac 901 atgactgttt taacaggaag acgccatgga cctctgcaga tgactggggc tccataccat 961 gataattgtc ttagctactt accattttct gggtcatcac agttaaaaag tcaccaaagt 1021 tcatttttcc tgtcccttcc ttatcaattt cacttatcat tttcttaatt tcttctttct 1081 tgggttcaaa gcccagggcc ctcattgcca cctataaaga aaacaggatc gtctcaataa 1141 gcacatctaa agtagcagga gcaaaaataa cagtgttgag agcattaagc cagcatctga 1201 tgtggtcatg ttaatatgat tggggaacac tatgacattg atctaattga tatttgcaat 1261 caattttatt gaataatact caaagtaagt aaagacagga aaacagcaca gtacaacctc 1321 ccctgttact tactgcaggg agaagagggc aaaaggggta gagaggtgat agggagatga 1381 caaggaagag gcagaggaaa ggaagggcag tacggcagga atgcacagag cttgccttca 1441 gttctttaac atctatggtg ccagttccat ccgcatcgaa aagatcaaaa gcttcccgga 1501 tctcctgctt ttgctcttca gtaagctcag gcttaggact cattcttttt cgctgagaac 1561 ttgatgccat gtttgccttt ttgaaattgg agcttctcag gccgctcttg tacagctcgt 1621 ccatgccatt aagtttgtgc cccagtttgc tagggaggtc gcagtatctg gccacagcca 1681 cctcgtgctg ctcgacgtag gtctctttgt cggcctcctt gattctttcc agtctgtggt 1741 ccacatagta gacgccgggc atcttgaggt tcttagcggg tttcttggat ctgtatgtgg 1801 tcttgaagtt gcagatcagg tggcccccgc ccacgagctt cagggccatg tcggttctgc 1861 cttccaggcc gccgtcagcg gggtacagca tctcggtgtt ggcctcccag ccgagtgttt 1921 tcttctgcat cacagggccg ttggatggga agttcacccc tctgatcttg acgttgtaga 1981 tgaggcagcc gtcctggagg ctggtgtcct gggtagcggt cagcacgccc ccgtcttcgt 2041 atgtggtgac tctctcccat gtgaagccct cagggaagga ctgcttaaag aaatcgggga 2101 tgccctgggt gtggttgatg aaggttctgc tgccgtacat gaagctggta gccaggatgt 2161 cgaaggcgaa ggggagaggg ccgccctcga ccaccttgat tctcatggtc tgggtgccct 2221 cgtagggctt gccttcgccc tcggatgtgc acttgaagtg gtggttgttc acggtgccct 2281 ccatgtacag cttcatgtgc atgttctcct taatcagctc ttcgccctta gacacggcct 2341 ttatatgtta tgcaatacac aagcacaata atatatcata cagtccatat ttacataaca 2401 tagttacaga cacccatatg tatttttaaa gtcatttata taaccaattc caaagctttg 2461 cttaaaaaga aagataacag ggtttactgg caagaacaat gaatctttcc atgaacttct 2521 acccctccct tttctttact gttgtttcta gaagcaggga gatcacttgt tcagtaagga 2581 cacctcttac tgaaaatcat cccaaaagag agccagtcaa gaagatgact gtgaataact 2641 agagataatc agctcttccc gaacgtgggg atcctggttc tgaaaacagt cttacgatgc 2701 ccactcccag tactcccctt cccagtaagt actccgactt cccaattact tctagatttg 2761 gccaaatatt agtaagttgt taagatgtat attgtgtcag taccgagtaa taaaactcaa 2821 tccatctggc accttcgtgc cttctccaag aagctaaagc atcccctgaa tgcccctgta 2881 tttaaattgt aaaaaacatt gtaacagaac acgatacacc ttttcccagc tgtgtgtgga 2941 aattttggca gaatgaggag gccagggggc caaggtctga gagagtcagg ggcctgtgag 3001 ggattcgagt cttttttgcc cagggaggtt caccgagggg atcagggaat gcaaggggag 3061 cccgaggggc gggtgtgggc ctgttgtggg tgaagaggag gcagcagcga agcgcaggcc 3121 cagtaggttg gttagcgtct gtaccctact ctcccgggaa agagctcaaa ggaaagactg 3181 gtgcgctaaa gccgagaaag gcgcaggaga ggcggcgtgg ccgagctggg gcctgggaag 3241 ggctcagccg ctgggcgtgg ggcgcgacag agcggcagca ctcaccatag ccaaaggagt 3301 ccgctgccgg ttgttaggca accgacgtgt acactatccc aatacgcgtc aattcactgg 3361 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg 3421 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt 3481 cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc 3541 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg 3601 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc 3661 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga 3721 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt 3781 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa 3841 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca 3901 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc 3961 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc 4021 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt 4081 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt 4141 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg 4201 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact 4261 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg 4321 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga 4381 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg 4441 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa 4501 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac 4561 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc 4621 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca 4681 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga 4741 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta 4801 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc 4861 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc 4921 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt 4981 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 5041 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 5101 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact 5161 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 5221 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 5281 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 5341 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 5401 ggagaaaggc ggaca //