LOCUS Exported 3979 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION For in vitro transcription of a human NODAL splice variant open reading frame RNA. ACCESSION . VERSION . KEYWORDS NODALvar-T7TS SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3979) AUTHORS Findlay SD, Bilyk O, Lypka K, Waskiewicz AJ, Postovit L TITLE Genetically regulated human NODAL splice variants are differentially post-transcriptionally processed and functionally distinct JOURNAL bioRxiv 276170 REFERENCE 2 (bases 1 to 3979) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3979 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 54..71 /label=L4440 /note="L4440 vector, forward primer" protein_bind 188..209 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 224..254 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 262..278 /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 267..289 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 286..302 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 286..302 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 321..340 /label=T7 /note="T7 promoter, forward primer" promoter 321..339 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" 5'UTR 352..394 /label=Xenopus globin 5'-UTR /note="translational enhancer from the 5'-UTR of the major beta-globin gene of Xenopus laevis" 3'UTR 1451..1576 /label=Xenopus globin 3'-UTR /note="3'-UTR of the major beta-globin gene of Xenopus laevis" promoter complement(1639..1657) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(1640..1657) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(1665..1682) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(1665..1681) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1674..1696) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(1890..1909) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2009..2031 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(2069..2087) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 2155..2259 /gene="bla" /label=AmpR promoter CDS 2260..3120 /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(2478..2497) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3291..3879 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3780..3799 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt 61 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc 121 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca 181 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc 241 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg 301 accatgatta cgccaagctc taatacgact cactataggg agacaagctt gcttgttctt 361 tttgcagaag ctcagaataa acgctcaact ttggcagatc tatgcacgcc cactgcctgc 421 ccttccttct gcacgcctgg tgggccctac tccaggcggg tgctgcgacg gtggccactg 481 cgctcctgcg tacgcggggg cagccctcgt cgccatcccc tctggcgtac atgctgagcc 541 tctaccgcga cccgctgccg agggcagaca tcatccgcag cctacaggca gaagatgtgg 601 cagtggatgg gcagaactgg acgtttgctt ttgacttctc cttcctgagc caacaagagg 661 atctggcatg ggctgagctc cggctgcagc tgtccagccc tgtggacctc cccactgagg 721 gctcacttgc cattgagatt ttccaccagc caaagcccga cacagagcag gcttcagaca 781 gctgcttaga gcggtttcag atggacctat tcactgtcac tttgtcccag gtcacctttt 841 ccttgggcag catggttttg gaggtgacca ggcctctctc caagtggctg aagcaccctg 901 gggccctgga gaagcagatg tccagggtag ctggagagtg ctggccgcgg ccccccacac 961 cgcctgccac caatgtgctc cttatgctct actccaacct ctcgcaggag cagaggcagc 1021 tgggtgggtc caccttgctg tgggaagccg agagctcctg gcgggcccag gagggacagc 1081 tgtcctggga gtggggcaag aggcaccgtc gacatcactt gccagacaga agtcaactgt 1141 gtcggaaggt caagttccag gtggacttca acctgatcgg atggggctcc tggatcatct 1201 accccaagca gtacaacgcc tatcgctgtg agggcgagtg tcctaatcct gttggggagg 1261 agtttcatcc gaccaaccat gcatacatcc aggtggccct gccctgctgt ccaaggtcat 1321 atgggaccaa aatgttttca ttttactcca tgaagtctgg aatgagaatt tcttggactt 1381 gcaatatatc ctccatgcca agcctcagag tctgctgaac tagtgactga ctaggatctg 1441 gttaccacta aaccagcctc aagaacaccc gaatggagtc tctaagctac ataataccaa 1501 cttacacttt acaaaatgtt gtcccccaaa atgtagccat tcgtatctgc tcctaataaa 1561 aagaaagttt cttcacattc taaaaacccc ccctgcaggt cgactctaga ggatccccgg 1621 gcgagctcga attcgtattc tatagtgtca cctaaatcca attcactggc cgtcgtttta 1681 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc 1741 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg 1801 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt 1861 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc 1921 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca 1981 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg 2041 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat 2101 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga 2161 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 2221 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt 2281 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg 2341 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg 2401 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg 2461 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag 2521 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca 2581 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg 2641 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc 2701 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg 2761 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg 2821 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac 2881 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg 2941 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg 3001 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact 3061 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa 3121 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt 3181 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag 3241 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 3301 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 3361 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 3421 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct 3481 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 3541 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 3601 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 3661 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 3721 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 3781 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 3841 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 3901 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct 3961 gattctgtgg ataaccgta //