LOCUS Exported 5265 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCMV-HA GCNF SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5265) AUTHORS Gu P, LeMenuet D, Chung AC, Mancini M, Wheeler DA, Cooney AJ TITLE Orphan nuclear receptor GCNF is required for the repression of pluripotency genes during retinoic acid-induced embryonic stem cell differentiation. JOURNAL Mol Cell Biol. 2005 Oct . 25(19):8507-19. PUBMED 16166633 REFERENCE 2 (bases 1 to 5265) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5265 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 27..330 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 331..534 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 485..505 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 531..555 /label=LNCX /note="Human CMV promoter, forward primer" intron 672..768 /label=SV40 intron /note="modified SV40 intron with splice donor and acceptor sites" CDS 832..858 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 2404..2538 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2428..2447) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2482..2501 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2649..2665) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2649..2665) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2662..2684) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2673..2689 /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(2697..2727) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2742..2763 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2880..2897) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3051..3639) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3131..3150) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3810..4670) /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 4433..4452 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4671..4775) /gene="bla" /label=AmpR promoter primer_bind 4843..4861 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(4899..4921) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 5021..5040 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5234..5256 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 5248..5265 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 5249..5265 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 gagttcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc 61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg 121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg 181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa 241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac 301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg 361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca 481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta 541 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac 601 cgggaccgat ccagcctccg gactctagag gatccggtac tagaggaact gaaaaaccag 661 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg 721 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa 781 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg ggcccaccat gtacccatac 841 gatgttccag attacgctct tatggccatg gaggcccgaa ttcgcatgga gcgggacgaa 901 cggccaccta gcggaggggg aggcggcggg ggctcggcgg ggttcctgga gccgcccgcc 961 gcgctccctc cgccgccgcg cagcggtttc tgtcaggatg aattggcaga gcttgatcca 1021 ggcactaatg gagagactga cagtttaaca cttggccaag gccatatacc tgtttccgtc 1081 ccagatgatc gagctgaaca acgaacctgt ctcatctgtg gggaccgcgc tacgggcttg 1141 cactatggga tcatctcctg cgagggctgc aaggggtttt tcaagaggag catttgcaac 1201 aaacgggtgt atcggtgcag tcgtgacaag aactgtgtca tgtcccggaa gcagaggaac 1261 agatgtcagt actgccgcct gctcaagtgt ctccagatgg gcatgaacag gaaggctatc 1321 agagaagatg gcatgcctgg aggccggaac aagagcattg gaccagtcca gatatcagaa 1381 gaagaaattg aaagaatcat gtctggacag gagtttgagg aagaagccaa tcactggagc 1441 aaccatggtg acagcgacca cagttcccct gggaacaggg cttcagagag caaccagccc 1501 tcaccaggct ccacactatc atccagtagg tctgtggaac taaatggatt catggcattc 1561 agggatcagt acatggggat gtcagtgcct ccacattatc aatacatacc acaccttttt 1621 agctattctg gccactcacc acttttgccc ccacaagctc gaagcctgga ccctcagtcc 1681 tacagtctga ttcatcagct gatgtcagcc gaagacctgg agccattggg cacacctatg 1741 ttgattgaag atgggtatgc tgtgacacag gcagaactgt ttgctctgct ttgccgcctg 1801 gccgacgagt tgctctttag gcagattgcc tggatcaaga agctgccttt cttctgcgag 1861 ctctcaatca aggattacac gtgcctcttg agctctacgt ggcaggagtt aatcctgctc 1921 tcctccctca cagtgtacag caagcagatc tttggggagc tggctgatgt cacagccaag 1981 tactcaccct ctgatgaaga actccacaga tttagtgatg aagggatgga ggtgattgaa 2041 cgactcatct acctatatca caagttccat cagctgaagg tcagcaacga ggagtacgca 2101 tgcatgaaag caattaactt cctggatcaa gatatcaggg gtctgaccag tgcctcacag 2161 ctggaacaac tgaacaagcg gtattggtac atttgtcagg atttcactga atataaatac 2221 acacatcagc caaaccgctt tcctgatctt atgatgtgct tgccagagat ccgatacatc 2281 gcaggcaaga tggtgaatgt gcccctggag cagctgcccc tcctctttaa ggtggtgctg 2341 cactcctgca agacaagtac ggtgaaggag tgacctgtgc cctcgaggta ccgcggccgc 2401 ggggatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag 2461 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata 2521 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg 2581 gaggtgtggg aggttttttc ggatcctcta gagtcgatct gcaggcatgc tagcttggcg 2641 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac 2701 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca 2761 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat 2821 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 2881 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 2941 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 3001 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 3061 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 3121 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 3181 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 3241 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 3301 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 3361 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 3421 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 3481 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 3541 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 3601 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 3661 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 3721 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 3781 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 3841 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 3901 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 3961 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 4021 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 4081 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 4141 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 4201 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 4261 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 4321 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 4381 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 4441 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 4501 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 4561 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 4621 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 4681 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 4741 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 4801 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg 4861 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg 4921 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg 4981 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta 5041 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc 5101 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc 5161 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta 5221 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagt //