LOCUS Exported 5229 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Provides pA sequences N-terminal to PGK-PacDeltaTK-pA cassette on Gateway entry vector. ACCESSION . VERSION . KEYWORDS pENTR-R2L3-pA-PGKpacDeltaTK-pA1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5229) TITLE Minoru Ko Lab plasmids REFERENCE 2 (bases 1 to 5229) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5229 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 92..109 /label=L4440 /note="L4440 vector, forward primer" terminator 268..295 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 387..473 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 522..544 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 537..553 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind complement(624..748) /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" polyA_signal 760..808 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" primer_bind 810..826 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 837..1336 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(872..895) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 1269..1288 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 1360..1956 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(1360..1379) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 1856..1876 /label=Puro-F /note="Puromycin resistance gene, forward primer" CDS 1963..2958 /codon_start=1 /product="truncated herpes simplex virus thymidine kinase (Salomon et al., 1995)" /label=-Delta-TK /note="confers sensitivity to nucleoside analogs such as acyclovir or ganciclovir" /translation="MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRV LGASETIANIYTTQHRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGS SHAPPPALTLIFDRHPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGAL PEDRHIDRLAKRQRPGERLDLAMLAAIRRVYGLLANTVRYLQCGGSWREDWGQLSGTAV PPQGAEPQSNAGPRPHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRSMHVFI LDYDQSPAGCRDALLQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN" protein_bind complement(3338..3433) /gene="mutant version of attL" /label=attL3 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(3470..3489) /label=T7 /note="T7 promoter, forward primer" promoter complement(3471..3489) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3494..3510) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3494..3510) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3612..3631) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 3623..4432 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(3690..3709) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 4579..5167 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5068..5087 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc 241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta 301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc 361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa 421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg 481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa 541 aacgacggcc agtcttaagc tcgggccctg cagctctaga gctcgaattc tacaggtcac 601 taataccatc taagtagttg gttcatagtg actgcatatg ttgtgtttta cagtattatg 661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt 721 ctcgttcaac tttcttgtac aaagtggtta attaaggatc ggcaataaaa agacagaata 781 aaacgcacgg gtgttgggtc gtttgttctg taaaacgacg gccagtgaat tctaccgggt 841 aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac 901 ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca 961 accggctccg ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa 1021 gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct 1081 cactagtctc gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg 1141 cagcggccaa tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg 1201 gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg 1261 aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc 1321 atctccgggc ctttcgaccg atcatcaagc ttgatcctca tgaccgagta caagcccacg 1381 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc 1441 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc 1501 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg 1561 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg 1621 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa 1681 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc 1741 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga 1801 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac 1861 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga 1921 ccgcgcacct ggtgcatgac ccgcaagccc ggtgccggat ccatgcccac gctactgcgg 1981 gtttatatag acggtcccca cgggatgggg aaaaccacca ccacgcaact gctggtggcc 2041 ctgggttcgc gcgacgatat cgtctacgta cccgagccga tgacttactg gcgggtgctg 2101 ggggcttccg agacaatcgc gaacatctac accacacaac accgcctcga ccagggtgag 2161 atatcggccg gggacgcggc ggtggtaatg acaagcgccc agataacaat gggcatgcct 2221 tatgccgtga ccgacgccgt tctggctcct catatcgggg gggaggctgg gagctcacat 2281 gccccgcccc cggccctcac cctcatcttc gaccgccatc ccatcgccgc cctcctgtgc 2341 tacccggccg cgcggtacct tatgggcagc atgacccccc aggccgtgct ggcgttcgtg 2401 gccctcatcc cgccgacctt gcccggcacc aacatcgtgc ttggggccct tccggaggac 2461 agacacatcg accgcctggc caaacgccag cgccccggcg agcggctgga cctggctatg 2521 ctggctgcga ttcgccgcgt ttacgggcta cttgccaata cggtgcggta tctgcagtgc 2581 ggcgggtcgt ggcgggagga ctggggacag ctttcgggga cggccgtgcc gccccagggt 2641 gccgagcccc agagcaacgc gggcccacga ccccatatcg gggacacgtt atttaccctg 2701 tttcgggccc ccgagttgct ggcccccaac ggcgacctgt ataacgtgtt tgcctgggcc 2761 ttggacgtct tggccaaacg cctccgttcc atgcacgtct ttatcctgga ttacgaccaa 2821 tcgcccgccg gctgccggga cgccctgctg caacttacct ccgggatggt ccagacccac 2881 gtcaccaccc ccggctccat accgacgata tgcgacctgg cgcgcacgtt tgcccgggag 2941 atgggggagg ctaactgagc tctagcttat cgactcgccg atagtggaaa ccgacgcccc 3001 agcactcgtc cgagggcaaa ggaatagtcg agaaattgat gatctattaa acaataaaga 3061 tgtccacatg gaagtttttc ctgtcatact ttgttaagaa gggtgagaac agagtaccta 3121 cattttgaat ggaaggattg gagctacggg ggtgggggtg gggtgggatt agataaatgc 3181 ctgctcttta ctgaaggctc tttactattg ctttatgata atgtttcata gttggatatc 3241 ataatttaaa caagcaaaac caaattaagg gccagctcat tcctcccact catgatctat 3301 agatcctcta gagtcgacct gcaggcatgc acccgggcaa ctttattata caaagttggc 3361 attataaaaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat 3421 aaaatcatta tttggagctc catggtagcg ttaacgcggc cgcgatatcc cctatagtga 3481 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg 3541 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat 3601 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt 3661 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca 3721 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa 3781 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct 3841 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg 3901 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga 3961 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc 4021 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg 4081 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt 4141 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac 4201 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat 4261 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg 4321 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa 4381 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt 4441 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc 4501 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga 4561 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac 4621 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt 4681 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc 4741 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat 4801 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag 4861 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc 4921 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag 4981 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac 5041 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg 5101 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct 5161 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc 5221 tcacatgtt //