LOCUS Exported 8765 bp ds-DNA circular SYN 26-JAN-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8765) AUTHORS Woltjen TITLE Direct Submission JOURNAL Exported Jan 26, 2018 from SnapGene 4.0.4 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8765 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 7..27 /label=MDSSR gene 103..1001 /gene="ENSG00000165704" /locus_tag="HPRT1" /label=ENSG00000165704 /note="hypoxanthine phosphoribosyltransferase 1 [Source:HGNC Symbol;Acc:5157]" primer_bind 103..122 /label=dna450 CDS 818..1001 /codon_start=2 /gene="ENSG00000165704" /label=ENSG00000165704 /label=ENSG00000165704(6) /note="transcript_id=ENST00000370796" /db_xref="UCSC:uc004exl.3" /db_xref="HPA:CAB012200" /db_xref="HPA:HPA006360" /db_xref="GO:GO:0004422" /db_xref="GO:GO:0005737" /db_xref="GO:GO:0006166" /db_xref="GO:GO:0009116" /db_xref="HGNC_transcript_name:HPRT1-201" /db_xref="IPI:IPI00873466.1" /db_xref="goslim_goa:GO:0005575" /db_xref="goslim_goa:GO:0005622" /db_xref="goslim_goa:GO:0005623" /db_xref="goslim_goa:GO:0005737" /db_xref="goslim_goa:GO:0009058" /db_xref="goslim_goa:GO:0034641" /db_xref="goslim_goa:GO:0044281" /protein_id="ENSP00000359832" /translation="TERLARDVMKEMGGHHIVALCVLKGGYKFFADLLDYIKALNRNSD RSIPMTVDFIRLKRYC" mRNA 818..1001 /gene="ENSG00000165704" /label=transcript_id=ENST00000298556 /label=ENSG00000165704(1) /note="transcript_id=ENST00000298556" misc_feature 968..976 /label=T microhomology primer_bind complement(971..1040) /label=Munich-IF-R misc_feature 985..993 /label=A microhomology misc_feature 989..1001 /label=MH-mutant misc_feature 989 /label=G>T (silent); AflII misc_feature 995 /label=312C>A; S104R misc_feature complement(1003..1025) /label=ps1 CDS 1026..1079 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" primer_bind 1026..1050 /label=T2A-pdtk-Fo CDS 1089..1685 /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 1650..1669 /label=dna930 CDS 1692..2687 /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" polyA_signal 2725..2949 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(2935..2950) /label=T2A-pdtk-Rev enhancer 2968..3347 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3349..3626 /label=chicken beta-actin promoter intron 3627..4642 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" protein_bind 4736..4760 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 4766..5476 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" protein_bind complement(5478..5502) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" misc_feature 5601..6059 /label=Rabbit b-globin pA polyA_site 5663..5718 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal" primer_bind 6093..6154 /label=Munich-IF-F misc_feature 6099..6121 /label=ps1 gene 6122..6488 /gene="ENSG00000165704" /locus_tag="HPRT1" /label=ENSG00000165704 /note="hypoxanthine phosphoribosyltransferase 1 [Source:HGNC Symbol;Acc:5157]" misc_feature 6122..6134 /label=MH-normal misc_feature 6122 /label=G>T (silent); AflII primer_bind complement(6465..6488) /label=dna411 primer_bind 6491..6507 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6528..6551) /label=pENTRfw rep_origin complement(6802..7390) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(7561..8421) /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" promoter complement(8422..8526) /gene="bla" /label=AmpR promoter ORIGIN 1 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc 61 acagatgcgt aaggagcgat cgcgaaaata ccgcatcagg cggtgcagtg cagcagaatg 121 atcacagttc actgcagtct caacctcctg ggttcaagcg atccttccac ctcagcctcc 181 tgagtagctg ggactatagg tgtgcgccac cactcccagc taatattttt attttgtaga 241 aacagggttc gccatgttac ccagcctccc aaagtgctgg gattacaggc atgagccact 301 ggcccagttt ctacagtctc tcttaatatt gtatattatc caagaaattt catttaatca 361 gaacctgcca gtctgatagg tgaaaatggt atcttgtttt tatttgcatt taaaaaaaat 421 tatgatagtg gtatgcttgg tttttttgaa ggtatcaaat tttttacctt atgaaacatg 481 agggcaaagg atgtgttacg tggaagattt aaaaaaaatt tttaatgcat ttttttgaga 541 caaggtcttg ctctattgtc caggctggag tgcagtggca caatcacagt tcactccagc 601 ctcaacatcc tgcactaaag tgattttccc acctcacctc tcaagtagct gggactacag 661 gtacatgcta ccatgcctgg ctaatttttt tttttttgca ggcatggggt ctcactatat 721 tgcccaggtt ggtgtggaag tttaatgact aagaggtgtt tgttataaag tttaatgtat 781 gaaactttct attaaattcc tgattttatt tctgtaggac tgaacgtctt gctcgagatg 841 tgatgaagga gatgggaggc catcacattg tagccctctg tgtgctcaag gggggctata 901 aattctttgc tgacctgctg gattacatca aagcactgaa tagaaatagt gatagatcca 961 ttcctatgac tgtagatttt atcagactta agagatattg taccaccggc aagctgcccg 1021 tgcccgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggcccta 1081 ggctcgagat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca 1141 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg 1201 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg 1261 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca 1321 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt 1381 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc 1441 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg 1501 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg 1561 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg 1621 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg 1681 gtgccggatc catgcccacg ctactgcggg tttatataga cggtccccac gggatgggga 1741 aaaccaccac cacgcaactg ctggtggccc tgggttcgcg cgacgatatc gtctacgtac 1801 ccgagccgat gacttactgg cgggtgctgg gggcttccga gacaatcgcg aacatctaca 1861 ccacacaaca ccgcctcgac cagggtgaga tatcggccgg ggacgcggcg gtggtaatga 1921 caagcgccca gataacaatg ggcatgcctt atgccgtgac cgacgccgtt ctggctcctc 1981 atatcggggg ggaggctggg agctcacatg ccccgccccc ggccctcacc ctcatcttcg 2041 accgccatcc catcgccgcc ctcctgtgct acccggccgc gcggtacctt atgggcagca 2101 tgacccccca ggccgtgctg gcgttcgtgg ccctcatccc gccgaccttg cccggcacca 2161 acatcgtgct tggggccctt ccggaggaca gacacatcga ccgcctggcc aaacgccagc 2221 gccccggcga gcggctggac ctggctatgc tggctgcgat tcgccgcgtt tacgggctac 2281 ttgccaatac ggtgcggtat ctgcagtgcg gcgggtcgtg gcgggaggac tggggacagc 2341 tttcggggac ggccgtgccg ccccagggtg ccgagcccca gagcaacgcg ggcccacgac 2401 cccatatcgg ggacacgtta tttaccctgt ttcgggcccc cgagttgctg gcccccaacg 2461 gcgacctgta taacgtgttt gcctgggcct tggacgtctt ggccaaacgc ctccgttcca 2521 tgcacgtctt tatcctggat tacgaccaat cgcccgccgg ctgccgggac gccctgctgc 2581 aacttacctc cgggatggtc cagacccacg tcaccacccc cggctccata ccgacgatat 2641 gcgacctggc gcgcacgttt gcccgggaga tgggggaggc taactgatct agagggcccg 2701 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc 2761 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 2821 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 2881 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg 2941 gctctatggg ctagcggtgg cggcctcgac attgattatt gactagttat taatagtaat 3001 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg 3061 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt 3121 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac 3181 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg 3241 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact 3301 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc 3361 ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat 3421 ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggg cgcgcgccag 3481 gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca 3541 atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct 3601 ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc 3661 gctccgcgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt 3721 gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct 3781 tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg 3841 ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc 3901 cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag 3961 tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg 4021 gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc 4081 ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc 4141 gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg 4201 gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct cgggggaggg 4261 gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca gccattgcct 4321 tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct gtgcggagcc 4381 gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc 4441 cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct 4501 ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg gacggggcag 4561 ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca 4621 tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca 4681 ttttggcaaa gaattcgcgg ccgcggtacc ggcgcgccgg atcctgcagg ttatcacaag 4741 tttgtacaaa aaagcaggct ccaccatggt gagcaagggc gaggaggaca acatggccat 4801 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt 4861 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa 4921 ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta 4981 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt 5041 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt 5101 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac 5161 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc 5221 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct 5281 gaaggacggc ggccactacg acgccgaggt caagaccacc tacaaggcca agaagcccgt 5341 gcagctgccc ggcgcctaca acgtcaacat caagctggac atcacctccc acaacgagga 5401 ctacaccatc gtggaacagt acgagcgcgc cgagggccgc cactccaccg gcggcatgga 5461 cgagctgtac aagtaagacc cagctttctt gtacaaagtg gtgataactc tagagaattc 5521 actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct 5581 cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc 5641 cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg 5701 aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag 5761 aatgagtatt tggtttagag tttggcaaca tatgccatat gctggctgcc atgaacaaag 5821 gtggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca 5881 tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc 5941 tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg 6001 actactccca gtcatagctg tccctcttct cttatgaaga tccctcgacc tgcagcccaa 6061 gcttggatcc ctcgagttaa ttaaatctag aagtcgacgg gcacgggcag cttgccggtg 6121 gtaagagcta ttgtgtgagt atatttaata tatgattctt tttagtggca acagtaggtt 6181 ttcttatatt ttctttgaat ctctgcaaac catacttgct ttcatttcac ttggttacag 6241 tgagattttt ctaacatatt cactagtact ttacatcaaa gccaatactg tttttttaaa 6301 actagtcacc ttggaggata tatacttatt ttacaggtgt gtgtggtttt ttaaataaac 6361 tccttttagg aattgctgtt gggacttggg atactttttt cactatacat actggtgaca 6421 gataccctct cttgagctac atcggtttgt ggggagtcaa aagtcctttg gagctaggtt 6481 tgacaaattt gtaaaacgac ggccagtgcg gccgcgaatt cggtacccta actaacagga 6541 agagtttgta ggggcccccc ctcgactgca ttaatgaatc ggccaacgcg cggggagagg 6601 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 6661 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 6721 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 6781 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 6841 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 6901 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 6961 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 7021 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 7081 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 7141 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 7201 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 7261 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 7321 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 7381 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 7441 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 7501 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 7561 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 7621 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 7681 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 7741 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 7801 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 7861 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 7921 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 7981 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 8041 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 8101 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 8161 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 8221 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 8281 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 8341 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 8401 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 8461 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 8521 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac 8581 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga 8641 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 8701 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 8761 gctta //