LOCUS Exported 7800 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression of zNanog Homeodomain. ACCESSION . VERSION . KEYWORDS pPB-CAG-zNanogHD-pgk-hph SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7800) AUTHORS Theunissen TW, Costa Y, Radzisheuskaya A, van Oosten AL, Lavial F, Pain B, Castro LF, Silva JC TITLE Reprogramming capacity of Nanog is functionally conserved in vertebrates and resides in a unique homeodomain. JOURNAL Development. 2011 Nov;138(22):4853-65. doi: 10.1242/dev.068775. PUBMED 22028025 REFERENCE 2 (bases 1 to 7800) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7800 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 383..658 /label=chicken beta-actin promoter intron 659..1670 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1594..1616 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1678..1697 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" protein_bind 1748..1772 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(2031..2055) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind complement(2076..2092) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 2100..2116 /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)." promoter complement(2124..2154) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2169..2190 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2250..2267) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2256..2480 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" protein_bind complement(2557..2590) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind complement(2813..2832) /label=T7 /note="T7 promoter, forward primer" promoter complement(2814..2832) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2842..2859) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2842..2858) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2851..2873) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3000..3455 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3087..3106) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3297..3318 /label=F1ori-F /note="F1 origin, forward primer" promoter 3481..3585 /gene="bla" /label=AmpR promoter CDS 3586..4446 /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(3804..3823) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4617..5205 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5106..5125 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5359..5376 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5493..5514 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5529..5559 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5567..5583 /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 5591..5607 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5626..5646 /label=T3 /note="T3 promoter, forward primer" promoter 5628..5646 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" repeat_region 5669..5703 /label=piggyBac left (5') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" CDS complement(6186..7211) /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" promoter complement(7230..7729) /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind 7671..7694 /label=MSCV-rev /note="Murine stem cell virus, reverse primer" protein_bind complement(7752..7785) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggactatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg ggtcgaggtg agccccacgt tctgcttcac tctccccatc 421 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg 481 atgggggcgg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg 541 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc 601 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg 661 agtcgctgcg ttgccttcgc cccgtgcccc gctccgcgcc gcctcgcgcc gcccgccccg 721 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg 781 ctgtaattag cgcttggttt aatgacggct cgtttctttt ctgtggctgc gtgaaagcct 841 taaagggctc cgggagggcc ctttgtgcgg gggggagcgg ctcggggggt gcgtgcgtgt 901 gtgtgtgcgt ggggagcgcc gcgtgcggcc cgcgctgccc ggcggctgtg agcgctgcgg 961 gcgcggcgcg gggctttgtg cgctccgcgt gtgcgcgagg ggagcgcggc cgggggcggt 1021 gccccgcggt gcgggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg 1081 gggggtgagc agggggtgtg ggcgcggcgg tcgggctgta acccccccct gcacccccct 1141 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtgcggg gcgtggcgcg 1201 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg 1261 cctcgggccg gggagggctc gggggagggg cgcggcggcc ccggagcgcc ggcggctgtc 1321 gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac 1381 ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta 1441 gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc 1501 gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg 1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg 1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca 1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcctcgac ggtatcgata 1741 agctatcaca agtttgtaca aaaaagcagg cttcaccatg cggcctccga ccctgccccg 1801 caagacccgg gctgcttttt ctgaagaaca gatgaatgct ttggtcaatc ggtttaatgt 1861 ccagagatac ctcacgcctg cagagatgaa aacgcttgct ggagcaacag gattaacata 1921 caagcaggta aaaacatggt ttcagaaccg caggatgaaa ctcaagaggc atcagagaga 1981 cagcagttgg atgactgaga gatatgttgt caatgctgta cccaactaag acccagcttt 2041 cttgtacaaa gtggtgatag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat 2101 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg 2161 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag 2221 tcgggaaacc tgtcgtgcca gcggatcagc ctcgactgtg ccttctagtt gccagccatc 2281 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct 2341 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 2401 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg 2461 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct cgagatccac 2521 tagttctagc ctcgaggcta gagcggccgc gtcgagataa cttcgtataa tgtatgctat 2581 acgaagttat gttttgttac tttatagaag aaattttgag tttttgtttt tttttaataa 2641 ataaataaac ataaataaat tgtttgttga atttattatt agtatgtaag tgtaaatata 2701 ataaaactta atatctattc aaattaataa ataaacgtca attttacgca tgattatctt 2761 taacgtacgt cacaatatga ttatctttct agggttaaag agctccaatt cgccctatag 2821 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc 2881 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag 2941 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga 3001 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc 3061 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac 3121 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag 3181 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc 3241 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg 3301 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata 3361 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa 3421 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg 3481 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga 3541 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat 3601 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca 3661 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc 3721 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca 3781 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg 3841 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca 3901 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata 3961 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag 4021 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg 4081 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca 4141 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta 4201 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct 4261 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca 4321 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag 4381 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat 4441 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt 4501 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa 4561 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 4621 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 4681 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 4741 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag 4801 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 4861 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 4921 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 4981 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 5041 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 5101 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 5161 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 5221 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 5281 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 5341 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc 5401 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc 5461 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca 5521 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa 5581 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac 5641 taaagggaac aaaagctggg taccttaacc ctagaaagat agtctgcgta aaattgacgc 5701 atgcattctt gaaatattgc tctctctttc taaatagcgc gaatccgtcg ctgtgcattt 5761 aggacatctc agtcgccgct tggagctccc gtgaggcgtg cttgtcaatg cggtaagtgt 5821 cactgatttt gaactataac gaccgcgtga gtcaaaatga cgcatgatta tcttttacgt 5881 gacttttaag atttaactca tacgataatt atattgttat ttcatgttct acttacgtga 5941 taacttatta tatatatatt ttcttgttat agatatccaa ctatgaaaca ttatcataaa 6001 gcaatagtaa agagccttca gtaaagagca ggcatttatc taatcccacc ccacccccac 6061 ccccgtagct ccaatccttc cattcaaaat gtaggtactc tgttctcacc cttcttaaca 6121 aagtatgaca ggaaaaaact tccatgtgga catctttatt gtttaataga tcatcaattt 6181 ctcgactatt cctttgccct cggacgagtg ctggggcgtc ggtttccact atcggcgagt 6241 acttctacac agccatcggt ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg 6301 acagtcccgg ctccggatcg gacgattgcg tcgcatcgac cctgcgccca agctgcatca 6361 tcgaaattgc cgtcaaccaa gctctgatag agttggtcaa gaccaatgcg gagcatatac 6421 gcccggagcc gcggcgatcc tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc 6481 tgctccatac aagccaacca cggcctccag aagaagatgt tggcgacctc gtattgggaa 6541 tccccgaaca tcgcctcgct ccagtcaatg accgctgtta tgcggccatt gtccgtcagg 6601 acattgttgg agccgaaatc cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc 6661 caaagcatca gctcatcgag agcctgcgcg acggacgcac tgacggtgtc gtccatcaca 6721 gtttgccagt gatacacatg gggatcagca atcgcgcata tgaaatcacg ccatgtagtg 6781 tattgaccga ttccttgcgg tccgaatggg ccgaacccgc tcgtctggct aagatcggcc 6841 gcagcgatcg catccatggc ctccgcgacc ggctgcagaa cagcgggcag ttcggtttca 6901 ggcaggtctt gcaacgtgac accctgtgca cggcgggaga tgcaataggt caggctctcg 6961 ctgaattccc caatgtcaag cacttccgga atcgggagcg cggccgatgc aaagtgccga 7021 taaacataac gatctttgta gaaaccatcg gcgcagctat ttacccgcag gacatatcca 7081 cgccctccta catcgaagct gaaagcacga gattcttcgc cctccgagag ctgcatcagg 7141 tcggagacgc tgtcgaactt ttcgatcaga aacttctcga cagacgtcgc ggtgagttca 7201 ggctttttca tggtggcggc tggatcggtc gaaaggcccg gagatgagga agaggagaac 7261 agcgcggcag acgtgcgctt ttgaagcgtg cgagaatgcc gggcctccgg aggaccttcg 7321 ggcgcccgcc ccgcccctga gcccgcccct gagcccgccc ccggacccac cccttcccag 7381 cctctgagcc cagaaagcga aggagcaaag ctgctattgg ccgctgcccc aaaggcctac 7441 ccgcttccat tgctcagcgg tgctgtccat ctgcacgaga ctagtgagac gtgctacttc 7501 catttgtcac gtcctgcacg acgcgagctg cggggcgggg gggaacttcc tgactagggg 7561 aggagtagaa ggtggcgcga aggggccacc aaagaacgga gccggttggc gcctaccggt 7621 ggatgtggaa tgtgtgcgag gccagaggcc acttgtgtag cgccaagtgc cagcggggct 7681 gctaaagcgc atgctccaga ctgccttggg aaaagcgcct cccctacccg gtagaattca 7741 tcgatgatat cataacttcg tataatgtat gctatacgaa gttatgcggc cctcgaggtc //