LOCUS Exported 8141 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pSicoR-Ef1a-mCh-Puro SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8141) AUTHORS Salomonis N, Schlieve CR, Pereira L, Wahlquist C, Colas A, Zambon AC, Vranizan K, Spindler MJ, Pico AR, Cline MS, Clark TA, Williams A, Blume JE, Samal E, Mercola M, Merrill BJ, Conklin BR TITLE Alternative splicing regulates mouse embryonic stem cell pluripotency and differentiation. JOURNAL Proc Natl Acad Sci U S A. 2010 Jun 8;107(23):10514-9. Epub 2010 May 24. PUBMED 20498046 REFERENCE 2 (bases 1 to 8141) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8141 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(56..75) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 247..626 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 628..826 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 781..801 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 844..1024 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1071..1196 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1695..1928 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2113..2157 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 2455..2572 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" protein_bind 2825..2858 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." promoter 3018..3229 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" LTR 3242..3510 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" regulatory 3548..3557 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3554..4261 /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" primer_bind complement(3699..3717) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 3967..3986 /label=mCherry-F /note="mCherry, forward primer" CDS 4268..4321 /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" CDS 4325..4921 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind 4818..4838 /label=Puro-F /note="Puromycin resistance gene, forward primer" protein_bind 4941..4974 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 5030..5618 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5083..5103) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5501..5512) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(5621..5637) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5622..5638) /label=pBluescriptKS /note="For pBluescript vector" LTR 6147..6327 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(6389..6977) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6469..6488) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7148..8008) /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 7771..7790 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8009..8113) /gene="bla" /label=AmpR promoter ORIGIN 1 gcttaagcgg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa 61 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg 121 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca 181 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata 241 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc 301 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac 361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa 421 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag 481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc 541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct 601 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg 661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt 721 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga 781 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt 841 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac 901 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg 961 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct 1021 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac 1081 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta 1141 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta 1201 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga 1261 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta 1321 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat 1381 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt 1441 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag 1501 agcaaaacaa aagtaagacc accgcacagc aagcggccgg ccgcgctgat cttcagacct 1561 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa 1621 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa 1681 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg 1741 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag 1801 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc 1861 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa 1921 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg 1981 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag 2041 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa 2101 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg 2161 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga 2221 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag 2281 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc 2341 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac 2401 ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa 2461 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac 2521 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta 2581 ttacagggac agcagagatc cagtttggtt agtaccgggc ccgctctaga gatccgacgc 2641 gccatctcta ggcccgcgcc ggccccctcg cacagacttg tgggagaagc tcggctactc 2701 ccctgccccg gttaatttgc atataatatt tcctagtaac tatagaggct taatgtgcga 2761 taaaagacag ataatctgtt ctttttaata ctagctacat tttacatgat aggcttggat 2821 ttctataact tcgtatagca tacattatac gaagttatac atgtcacaaa aggaaactca 2881 ccctaactgt aaagtaattg tgtgttttga gactataaat atcccttgga gaaaagcctt 2941 gttaacgcgc ggtgaccctc gagtactagg atccattagg cggccgcgga tctgcgatcg 3001 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg 3061 aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga 3121 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt 3181 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg 3241 aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg 3301 gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta 3361 ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc 3421 tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt 3481 ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagcgct 3541 accggtcgcc accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt 3601 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg 3661 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg 3721 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc 3781 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt 3841 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc 3901 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc 3961 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta 4021 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg 4081 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg 4141 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt 4201 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa 4261 gccccgggag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc 4321 tcgaaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt 4381 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga 4441 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga 4501 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga 4561 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg 4621 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga 4681 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg 4741 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct 4801 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc 4861 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta 4921 agaattcgtc gagggaccta ataacttcgt atagcataca ttatacgaag ttatacatgt 4981 ttaagggttc cggttccact aggtacaatt cgatatcaag cttatcgata atcaacctct 5041 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct 5101 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat 5161 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt 5221 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat 5281 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc 5341 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga 5401 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc 5461 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga 5521 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc 5581 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgatc 5641 gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt 5701 gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac 5761 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg 5821 ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct 5881 accacacaca aggctacttc cctgattggc agaactacac accagggcca gggatcagat 5941 atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag aaggtagaag 6001 aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat gggatggatg 6061 acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt catcacatgg 6121 cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc tgagcctggg 6181 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc 6241 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct 6301 tttagtcagt gtggaaaatc tctagcagca tgtgagcaaa aggccagcaa aaggccagga 6361 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 6421 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 6481 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 6541 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt 6601 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 6661 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 6721 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 6781 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg 6841 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 6901 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 6961 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 7021 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 7081 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 7141 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 7201 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 7261 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 7321 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 7381 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 7441 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 7501 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 7561 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 7621 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 7681 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 7741 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 7801 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 7861 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 7921 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 7981 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 8041 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 8101 taggggttcc gcgcacattt ccccgaaaag tgccacctga c //