LOCUS Exported 6811 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mutational dissection of the G rich region of the c-kit promoter. ACCESSION . VERSION . KEYWORDS pC-KIT5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6811) AUTHORS Ducani C., Bernardinelli G., Hogberg B., Keppler B.K., Terenzi A. TITLE Interplay of three G-quadruplex units in the KIT promoter JOURNAL J. Am. Chem. Soc. 2019 REFERENCE 2 (bases 1 to 6811) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6811 /organism="synthetic DNA construct" /mol_type="other DNA" regulatory 1276..1285 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1282..2217 /codon_start=1 /product="Renilla luciferase" /label=hRluc /note="human codon-optimized" /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ" polyA_signal 2276..2324 /note="synthetic polyadenylation signal" promoter 2333..3084 /label=HSV TK promoter /note="herpes simplex virus thymidine kinase promoter" regulatory 3114..3123 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3120..4772 /codon_start=1 /gene="luc+" /product="firefly luciferase" /label=luciferase /note="enhanced luc+ version of the luciferase gene" /db_xref="GI:42718124" /protein_id="AAS38485.1" /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" RBS 4079..4087 /label=Shine-Dalgarno sequence /note="full consensus sequence for ribosome-binding sites upstream of start codons in E. coli; complementary to a region in the 3' end of the 16S rRNA (Chen et al., 1994)" polyA_signal 4816..4937 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4827..4846) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 4881..4900 /label=SV40pA-R /note="SV40 polyA, reverse primer" promoter 5070..5174 /gene="bla" /label=AmpR promoter CDS 5175..6035 /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(5393..5412) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6206..6794 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6695..6714 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atggctcgac agatctgagg 61 gatgtcttat tagtttatca gaatgcatag ggaaagctca tctttcttgg agtaggtctt 121 ccaataagtg ttcaagccct gggtcttccc tcactacaat aataaacata cctgctctac 181 cctcctcaca aagctgttat cgtgagcatg aaaagacgag tcgctgtggg gaagaactat 241 taacccctat acagatactg ttgttgcttt ccgttcaagg atccaccatg agactgtggg 301 taaggcactt tctgggcagt gaaatcgggc ttaagagaat cattgtactt ttattttgct 361 cctgggaccg gcaagggcag ttaagagccc tagccctaaa actgtcgtct gttacagagg 421 cagagaaatc attactgggg ttggcagcgt tggggggagg ggggaaaacg tgtatgaaaa 481 cctgggctcc cagagcaaac ttcgccacgc gccctccatc gcccctgtct ccacacaagt 541 cccggcaggg gcttcttggg cggaggccac gccgaccaat aggaacccac tgtgttccta 601 caggttacga agcaggtgga gaaattgagc agaaacaatt agcgaaaccg ggctcagcct 661 ttaccgcggt gccaggagct cctaacaggc ctggagggga atgtcggggc tcaatttcct 721 aacgctcccc tccccatccc catgccacct ccacgagcag cggcgtccag cctcctcccg 781 cccgaacgtg ctcgaggggc gggcagtcga cctttattgt ctggggagca cctggcaggt 841 ggcgggcccg tgccctaacg tgtgcgtggt gcccagcttc acaaagcgag cgggcagcac 901 ctccttggtc cgggaacgcc tcagcctggc cgtccacatc ccaggggtgg aaaggtggag 961 agagaaaggg gctccggagt caagagcggg gagagagggc gcgcgccctc ctcctcccgg 1021 cgggcacagc cccccggcat taacacgtcg aaagagcagg ggccagacgc cgccgggaag 1081 aagcgagacc caaacaaacg cgaaaaagaa aagacgagaa gaaacgtgac cgacgcgcag 1141 agggagggcg ctgggaggag gggctgctgc tcgccgctcg cggctctggg ggctcggctt 1201 tgccgcgctc gctgcacttg ggcgagagct ggaacgtgga ccagagctcg gatcccatcg 1261 cagctaccgc ggctagccac catggcttcc aaggtgtacg accccgagca acgcaaacgc 1321 atgatcactg ggcctcagtg gtgggctcgc tgcaagcaaa tgaacgtgct ggactccttc 1381 atcaactact atgattccga gaagcacgcc gagaacgccg tgatttttct gcatggtaac 1441 gctgcctcca gctacctgtg gaggcacgtc gtgcctcaca tcgagcccgt ggctagatgc 1501 atcatccctg atctgatcgg aatgggtaag tccggcaaga gcgggaatgg ctcatatcgc 1561 ctcctggatc actacaagta cctcaccgct tggttcgagc tgctgaacct tccaaagaaa 1621 atcatctttg tgggccacga ctggggggct tgtctggcct ttcactactc ctacgagcac 1681 caagacaaga tcaaggccat cgtccatgct gagagtgtcg tggacgtgat cgagtcctgg 1741 gacgagtggc ctgacatcga ggaggatatc gccctgatca agagcgaaga gggcgagaaa 1801 atggtgcttg agaataactt cttcgtcgag accatgctcc caagcaagat catgcggaaa 1861 ctggagcctg aggagttcgc tgcctacctg gagccattca aggagaaggg cgaggttaga 1921 cggcctaccc tctcctggcc tcgcgagatc cctctcgtta agggaggcaa gcccgacgtc 1981 gtccagattg tccgcaacta caacgcctac cttcgggcca gcgacgatct gcctaagatg 2041 ttcatcgagt ccgaccctgg gttcttttcc aacgctattg tcgagggagc taagaagttc 2101 cctaacaccg agttcgtgaa ggtgaagggc ctccacttca gccaggagga cgctccagat 2161 gaaatgggta agtacatcaa gagcttcgtg gagcgcgtgc tgaagaacga gcagtaattc 2221 taggcgatcg ctcgagcccg ggaattcgtt taaacctaga gcggccgctg gccgcaataa 2281 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaggatc taaatgagtc 2341 ttcggacctc gcgggggccg cttaagcggt ggttagggtt tgtctgacgc ggggggaggg 2401 ggaaggaacg aaacactctc attcggaggc ggctcggggt ttggtcttgg tggccacggg 2461 cacgcagaag agcgccgcga tcctcttaag cacccccccg ccctccgtgg aggcgggggt 2521 ttggtcggcg ggtggtaact ggcgggccgc tgactcgggc gggtcgcgcg ccccagagtg 2581 tgaccttttc ggtctgctcg cagacccccg ggcggcgccg ccgcggcggc gacgggctcg 2641 ctgggtccta ggctccatgg ggaccgtata cgtggacagg ctctggagca tccgcacgac 2701 tgcggtgata ttaccggaga ccttctgcgg gacgagccgg gtcacgcggc tgacgcggag 2761 cgtccgttgg gcgacaaaca ccaggacggg gcacaggtac actatcttgt cacccggagg 2821 cgcgagggac tgcaggagct tcagggagtg gcgcagctgc ttcatccccg tggcccgttg 2881 ctcgcgtttg ctggcggtgt ccccggaaga aatatatttg catgtcttta gttctatgat 2941 gacacaaacc ccgcccagcg tcttgtcatt ggcgaattcg aacacgcaga tgcagtcggg 3001 gcggcgcggt cccaggtcca cttcgcatat taaggtgacg cgtgtggcct cgaacaccga 3061 gcgaccctgc agcgacccgc ttaaaagctt ggcattccgg tactgttggt aaagccacca 3121 tggccgatgc taagaacatt aagaagggcc ctgctccctt ctaccctctg gaggatggca 3181 ccgctggcga gcagctgcac aaggccatga agaggtatgc cctggtgcct ggcaccattg 3241 ccttcaccga tgcccacatt gaggtggaca tcacctatgc cgagtacttc gagatgtctg 3301 tgcgcctggc cgaggccatg aagaggtacg gcctgaacac caaccaccgc atcgtggtgt 3361 gctctgagaa ctctctgcag ttcttcatgc cagtgctggg cgccctgttc atcggagtgg 3421 ccgtggcccc tgctaacgac atttacaacg agcgcgagct gctgaacagc atgggcattt 3481 ctcagcctac cgtggtgttc gtgtctaaga agggcctgca gaagatcctg aacgtgcaga 3541 agaagctgcc tatcatccag aagatcatca tcatggactc taagaccgac taccagggct 3601 tccagagcat gtacacattc gtgacatctc atctgcctcc tggcttcaac gagtacgact 3661 tcgtgccaga gtctttcgac agggacaaaa ccattgccct gatcatgaac agctctgggt 3721 ctaccggcct gcctaagggc gtggccctgc ctcatcgcac cgcctgtgtg cgcttctctc 3781 acgcccgcga ccctattttc ggcaaccaga tcatccccga caccgctatt ctgagcgtgg 3841 tgccattcca ccacggcttc ggcatgttca ccaccctggg ctacctgatt tgcggctttc 3901 gggtggtgct gatgtaccgc ttcgaggagg agctgttcct gcgcagcctg caagactaca 3961 aaattcagtc tgccctgctg gtgccaaccc tgttcagctt cttcgctaag agcaccctga 4021 tcgacaagta cgacctgtct aacctgcacg agattgcctc tggcggcgcc ccactgtcta 4081 aggaggtggg cgaagccgtg gccaagcgct ttcatctgcc aggcatccgc cagggctacg 4141 gcctgaccga gacaaccagc gccattctga ttaccccaga gggcgacgac aagcctggcg 4201 ccgtgggcaa ggtggtgcca ttcttcgagg ccaaggtggt ggacctggac accggcaaga 4261 ccctgggagt gaaccagcgc ggcgagctgt gtgtgcgcgg ccctatgatt atgtccggct 4321 acgtgaataa ccctgaggcc acaaacgccc tgatcgacaa ggacggctgg ctgcactctg 4381 gcgacattgc ctactgggac gaggacgagc acttcttcat cgtggaccgc ctgaagtctc 4441 tgatcaagta caagggctac caggtggccc cagccgagct ggagtctatc ctgctgcagc 4501 accctaacat tttcgacgcc ggagtggccg gcctgcccga cgacgatgcc ggcgagctgc 4561 ctgccgccgt cgtcgtgctg gaacacggca agaccatgac cgagaaggag atcgtggact 4621 atgtggccag ccaggtgaca accgccaaga agctgcgcgg cggagtggtg ttcgtggacg 4681 aggtgcccaa gggcctgacc ggcaagctgg acgcccgcaa gatccgcgag atcctgatca 4741 aggctaagaa aggcggcaag atcgccgtgt aataattcta gagtcggggc ggccggccgc 4801 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt 4861 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa 4921 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg 4981 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg 5041 atccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa 5101 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 5161 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg 5221 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag 5281 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg 5341 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg 5401 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt 5461 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga 5521 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac 5581 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc 5641 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc 5701 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac 5761 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag 5821 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg 5881 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta 5941 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg 6001 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata 6061 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt 6121 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 6181 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 6241 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 6301 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag 6361 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 6421 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 6481 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 6541 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 6601 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 6661 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 6721 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 6781 ggagcctatg gaaaaacgcc agcaacgcgg c //