LOCUS       sequence-229522-        6811 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mutational dissection of the G rich region of the c-kit promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6811)
  AUTHORS   Ducani C, Bernardinelli G, Hogberg B, Keppler BK, Terenzi A
  TITLE     Interplay of Three G-Quadruplex Units in the KIT Promoter.
  JOURNAL   J Am Chem Soc. 2019 Jul 3;141(26):10205-10213. doi: 
            10.1021/jacs.8b12753. Epub 2019 Jun 20.
  PUBMED    31244182
REFERENCE   2  (bases 1 to 6811)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/jacs.8b12753"; journalName: "J Am Chem Soc"; date: 
            "2019-07-3- 3"; volume: "141"; issue: "26"; pages: "10205-10213"
COMMENT     Sequence Label: pC-KIT7
FEATURES             Location/Qualifiers
     source          1..6811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     326..345
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      1718..1727
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1724..2659
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2718..2766
                     /note="synthetic polyadenylation signal"
     promoter        2775..3526
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      3556..3565
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3562..5214
                     /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             4521..4529
                     /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    5258..5379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5269..5288)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5323..5342
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5512..5616
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5617..5685,5686..6477)
                     /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(5835..5854)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6648..6811,1..425)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
       61 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      121 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      181 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      241 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      301 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      361 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      421 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      481 acatggctcg acagatctga gggatgtctt attagtttat cagaatgcat agggaaagct
      541 catctttctt ggagtaggtc ttccaataag tgttcaagcc ctgggtcttc cctcactaca
      601 ataataaaca tacctgctct accctcctca caaagctgtt atcgtgagca tgaaaagacg
      661 agtcgctgtg gggaagaact attaacccct atacagatac tgttgttgct ttccgttcaa
      721 ggatccacca tgagactgtg ggtaaggcac tttctgggca gtgaaatcgg gcttaagaga
      781 atcattgtac ttttattttg ctcctgggac cggcaagggc agttaagagc cctagcccta
      841 aaactgtcgt ctgttacaga ggcagagaaa tcattactgg ggttggcagc gttgggggga
      901 ggggggaaaa cgtgtatgaa aacctgggct cccagagcaa acttcgccac gcgccctcca
      961 tcgcccctgt ctccacacaa gtcccggcag gggcttcttg ggcggaggcc acgccgacca
     1021 ataggaaccc actgtgttcc tacaggttac gaagcaggtg gagaaattga gcagaaacaa
     1081 ttagcgaaac cgggctcagc ctttaccgcg gtgccaggag ctcctaacag gcctggaggg
     1141 gaatgtcggg gctcaatttc ctaacgctcc cctccccatc cccatgccac ctccacgagc
     1201 agcggcgtcc agcctcctcc cgcccgaacg tgctcgaggg gcgggcagtc gacctttatt
     1261 gtctggggag cacctggcag gtggcgggcc cgtgccctaa cgtgtgcgtg gtgcccagct
     1321 tcacaaagcg agcgggcagc acctccttgg tccgggaacg cctcagcctg gccgtccaca
     1381 tcccaggggt ggaaaggtgg agagagaaag gggctccgga gtcaagagcg gggagagagg
     1441 gcgcgcgccc tcctcctccc ggcgggcaca gccccccggc attaacacgt cgaaagagca
     1501 ggggccagac gccgccggga agaagcgaga cccgggcggg cgcgagggag gggagacgag
     1561 aagaaacgtg accgacgcgc agaaagagaa cgctaagagg aggaactgct gctcgccgct
     1621 cgcggctctg ggggctcggc tttgccgcgc tcgctgcact tgggcgagag ctggaacgtg
     1681 gaccagagct cggatcccat cgcagctacc gcggctagcc accatggctt ccaaggtgta
     1741 cgaccccgag caacgcaaac gcatgatcac tgggcctcag tggtgggctc gctgcaagca
     1801 aatgaacgtg ctggactcct tcatcaacta ctatgattcc gagaagcacg ccgagaacgc
     1861 cgtgattttt ctgcatggta acgctgcctc cagctacctg tggaggcacg tcgtgcctca
     1921 catcgagccc gtggctagat gcatcatccc tgatctgatc ggaatgggta agtccggcaa
     1981 gagcgggaat ggctcatatc gcctcctgga tcactacaag tacctcaccg cttggttcga
     2041 gctgctgaac cttccaaaga aaatcatctt tgtgggccac gactgggggg cttgtctggc
     2101 ctttcactac tcctacgagc accaagacaa gatcaaggcc atcgtccatg ctgagagtgt
     2161 cgtggacgtg atcgagtcct gggacgagtg gcctgacatc gaggaggata tcgccctgat
     2221 caagagcgaa gagggcgaga aaatggtgct tgagaataac ttcttcgtcg agaccatgct
     2281 cccaagcaag atcatgcgga aactggagcc tgaggagttc gctgcctacc tggagccatt
     2341 caaggagaag ggcgaggtta gacggcctac cctctcctgg cctcgcgaga tccctctcgt
     2401 taagggaggc aagcccgacg tcgtccagat tgtccgcaac tacaacgcct accttcgggc
     2461 cagcgacgat ctgcctaaga tgttcatcga gtccgaccct gggttctttt ccaacgctat
     2521 tgtcgaggga gctaagaagt tccctaacac cgagttcgtg aaggtgaagg gcctccactt
     2581 cagccaggag gacgctccag atgaaatggg taagtacatc aagagcttcg tggagcgcgt
     2641 gctgaagaac gagcagtaat tctaggcgat cgctcgagcc cgggaattcg tttaaaccta
     2701 gagcggccgc tggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     2761 tgtgtgagga tctaaatgag tcttcggacc tcgcgggggc cgcttaagcg gtggttaggg
     2821 tttgtctgac gcggggggag ggggaaggaa cgaaacactc tcattcggag gcggctcggg
     2881 gtttggtctt ggtggccacg ggcacgcaga agagcgccgc gatcctctta agcacccccc
     2941 cgccctccgt ggaggcgggg gtttggtcgg cgggtggtaa ctggcgggcc gctgactcgg
     3001 gcgggtcgcg cgccccagag tgtgaccttt tcggtctgct cgcagacccc cgggcggcgc
     3061 cgccgcggcg gcgacgggct cgctgggtcc taggctccat ggggaccgta tacgtggaca
     3121 ggctctggag catccgcacg actgcggtga tattaccgga gaccttctgc gggacgagcc
     3181 gggtcacgcg gctgacgcgg agcgtccgtt gggcgacaaa caccaggacg gggcacaggt
     3241 acactatctt gtcacccgga ggcgcgaggg actgcaggag cttcagggag tggcgcagct
     3301 gcttcatccc cgtggcccgt tgctcgcgtt tgctggcggt gtccccggaa gaaatatatt
     3361 tgcatgtctt tagttctatg atgacacaaa ccccgcccag cgtcttgtca ttggcgaatt
     3421 cgaacacgca gatgcagtcg gggcggcgcg gtcccaggtc cacttcgcat attaaggtga
     3481 cgcgtgtggc ctcgaacacc gagcgaccct gcagcgaccc gcttaaaagc ttggcattcc
     3541 ggtactgttg gtaaagccac catggccgat gctaagaaca ttaagaaggg ccctgctccc
     3601 ttctaccctc tggaggatgg caccgctggc gagcagctgc acaaggccat gaagaggtat
     3661 gccctggtgc ctggcaccat tgccttcacc gatgcccaca ttgaggtgga catcacctat
     3721 gccgagtact tcgagatgtc tgtgcgcctg gccgaggcca tgaagaggta cggcctgaac
     3781 accaaccacc gcatcgtggt gtgctctgag aactctctgc agttcttcat gccagtgctg
     3841 ggcgccctgt tcatcggagt ggccgtggcc cctgctaacg acatttacaa cgagcgcgag
     3901 ctgctgaaca gcatgggcat ttctcagcct accgtggtgt tcgtgtctaa gaagggcctg
     3961 cagaagatcc tgaacgtgca gaagaagctg cctatcatcc agaagatcat catcatggac
     4021 tctaagaccg actaccaggg cttccagagc atgtacacat tcgtgacatc tcatctgcct
     4081 cctggcttca acgagtacga cttcgtgcca gagtctttcg acagggacaa aaccattgcc
     4141 ctgatcatga acagctctgg gtctaccggc ctgcctaagg gcgtggccct gcctcatcgc
     4201 accgcctgtg tgcgcttctc tcacgcccgc gaccctattt tcggcaacca gatcatcccc
     4261 gacaccgcta ttctgagcgt ggtgccattc caccacggct tcggcatgtt caccaccctg
     4321 ggctacctga tttgcggctt tcgggtggtg ctgatgtacc gcttcgagga ggagctgttc
     4381 ctgcgcagcc tgcaagacta caaaattcag tctgccctgc tggtgccaac cctgttcagc
     4441 ttcttcgcta agagcaccct gatcgacaag tacgacctgt ctaacctgca cgagattgcc
     4501 tctggcggcg ccccactgtc taaggaggtg ggcgaagccg tggccaagcg ctttcatctg
     4561 ccaggcatcc gccagggcta cggcctgacc gagacaacca gcgccattct gattacccca
     4621 gagggcgacg acaagcctgg cgccgtgggc aaggtggtgc cattcttcga ggccaaggtg
     4681 gtggacctgg acaccggcaa gaccctggga gtgaaccagc gcggcgagct gtgtgtgcgc
     4741 ggccctatga ttatgtccgg ctacgtgaat aaccctgagg ccacaaacgc cctgatcgac
     4801 aaggacggct ggctgcactc tggcgacatt gcctactggg acgaggacga gcacttcttc
     4861 atcgtggacc gcctgaagtc tctgatcaag tacaagggct accaggtggc cccagccgag
     4921 ctggagtcta tcctgctgca gcaccctaac attttcgacg ccggagtggc cggcctgccc
     4981 gacgacgatg ccggcgagct gcctgccgcc gtcgtcgtgc tggaacacgg caagaccatg
     5041 accgagaagg agatcgtgga ctatgtggcc agccaggtga caaccgccaa gaagctgcgc
     5101 ggcggagtgg tgttcgtgga cgaggtgccc aagggcctga ccggcaagct ggacgcccgc
     5161 aagatccgcg agatcctgat caaggctaag aaaggcggca agatcgccgt gtaataattc
     5221 tagagtcggg gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     5281 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     5341 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     5401 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     5461 aaatgtggta aaatcgataa ggatccaggt ggcacttttc ggggaaatgt gcgcggaacc
     5521 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5581 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5641 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5701 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5761 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5821 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5881 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5941 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6001 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6061 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6121 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6181 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6241 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6301 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6361 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6421 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6481 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6541 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6601 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6661 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6721 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6781 ataccaaata ctgttcttct agtgtagccg t
//
