LOCUS Exported 8181 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pSicoR-Ef1a-mCh-Puro-GFPi SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8181) 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 8181) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8181 /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 3058..3269 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" LTR 3282..3550 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" regulatory 3588..3597 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3594..4301 /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(3739..3757) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 4007..4026 /label=mCherry-F /note="mCherry, forward primer" CDS 4308..4361 /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 4365..4961 /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 4858..4878 /label=Puro-F /note="Puromycin resistance gene, forward primer" protein_bind 4981..5014 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 5070..5658 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5123..5143) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5541..5552) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(5661..5677) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5662..5678) /label=pBluescriptKS /note="For pBluescript vector" LTR 6187..6367 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(6429..7017) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6509..6528) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7188..8048) /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 7811..7830 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8049..8153) /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 gtttacagcc acaacgtcta tatttcaaga gaatatagac gttgtggctg tttttttctc 3001 gagtactagg atccattagg cggccgcgga tctgcgatcg ctccggtgcc cgtcagtggg 3061 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg 3121 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc 3181 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt 3241 ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc atctctcctt 3301 cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc gttctgccgc 3361 ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt 3421 cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc cggctctcca 3481 cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct gttctgcgcc 3541 gttacagatc caagctgtga ccggcgccta cgctagcgct accggtcgcc accatggtga 3601 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca 3661 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct 3721 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct 3781 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg 3841 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga 3901 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt 3961 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga 4021 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga 4081 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca 4141 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca 4201 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg 4261 agggccgcca ctccaccggc ggcatggacg agctgtacaa gccccgggag ggcagaggaa 4321 gtcttctaac atgcggtgac gtggaggaga atcccggccc tcgaaccgag tacaagccca 4381 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt 4441 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca 4501 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg 4561 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg 4621 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc 4681 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca 4741 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg 4801 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca 4861 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag 4921 gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta agaattcgtc gagggaccta 4981 ataacttcgt atagcataca ttatacgaag ttatacatgt ttaagggttc cggttccact 5041 aggtacaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa 5101 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa 5161 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat 5221 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt 5281 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc 5341 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc 5401 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg 5461 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga 5521 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc 5581 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc 5641 tttgggccgc ctccccgcat cgataccgtc gacctcgatc gagacctaga aaaacatgga 5701 gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa 5761 gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac 5821 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt 5881 cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc 5941 cctgattggc agaactacac accagggcca gggatcagat atccactgac ctttggatgg 6001 tgctacaagc tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac 6061 acccgcttgt tacaccctgt gagcctgcat gggatggatg acccggagag agaagtatta 6121 gagtggaggt ttgacagccg cctagcattt catcacatgg cccgagagct gcatccggac 6181 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg 6241 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt 6301 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc 6361 tctagcagca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 6421 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 6481 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 6541 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 6601 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc 6661 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 6721 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 6781 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 6841 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag 6901 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 6961 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 7021 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 7081 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 7141 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 7201 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 7261 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 7321 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 7381 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 7441 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 7501 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc 7561 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc 7621 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca 7681 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag 7741 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg 7801 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa 7861 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 7921 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga 7981 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga 8041 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 8101 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 8161 ccccgaaaag tgccacctga c //