LOCUS       sequence-229528-        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-KIT2
FEATURES             Location/Qualifiers
     source          1..6811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1260..1269
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1266..2201
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2260..2308
                     /note="synthetic polyadenylation signal"
     promoter        2317..3068
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      3098..3107
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3104..4756
                     /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             4063..4071
                     /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    4800..4921
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4811..4830)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4865..4884
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5054..5158
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5159..5227,5228..6019)
                     /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(5377..5396)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6190..6778
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6679..6698
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccttttgct ggccttttgc tcacatggct cgacagatct gagggatgtc ttattagttt
       61 atcagaatgc atagggaaag ctcatctttc ttggagtagg tcttccaata agtgttcaag
      121 ccctgggtct tccctcacta caataataaa catacctgct ctaccctcct cacaaagctg
      181 ttatcgtgag catgaaaaga cgagtcgctg tggggaagaa ctattaaccc ctatacagat
      241 actgttgttg ctttccgttc aaggatccac catgagactg tgggtaaggc actttctggg
      301 cagtgaaatc gggcttaaga gaatcattgt acttttattt tgctcctggg accggcaagg
      361 gcagttaaga gccctagccc taaaactgtc gtctgttaca gaggcagaga aatcattact
      421 ggggttggca gcgttggggg gaggggggaa aacgtgtatg aaaacctggg ctcccagagc
      481 aaacttcgcc acgcgccctc catcgcccct gtctccacac aagtcccggc aggggcttct
      541 tgggcggagg ccacgccgac caataggaac ccactgtgtt cctacaggtt acgaagcagg
      601 tggagaaatt gagcagaaac aattagcgaa accgggctca gcctttaccg cggtgccagg
      661 agctcctaac aggcctggag gggaatgtcg gggctcaatt tcctaacgct cccctcccca
      721 tccccatgcc acctccacga gcagcggcgt ccagcctcct cccgcccgaa cgtgctcgag
      781 gggcgggcag tcgaccttta ttgtctgggg agcacctggc aggtggcggg cccgtgccct
      841 aacgtgtgcg tggtgcccag cttcacaaag cgagcgggca gcacctcctt ggtccgggaa
      901 cgcctcagcc tggccgtcca catcccaggg gtggaaaggt ggagagagaa aggggctccg
      961 gagtcaagag cggggagaga gggcgcgcgc cctcctcctc ccggcgggca cagccccccg
     1021 gcattaacac gtcgaaagag caggggccag acgccgccgg gaagaagcga gacccaaaca
     1081 aacgcgaaaa agaaaaggcg aggaggggcg tggccggcgc gcagagggag ggcgctggga
     1141 ggaggggctg ctgctcgccg ctcgcggctc tgggggctcg gctttgccgc gctcgctgca
     1201 cttgggcgag agctggaacg tggaccagag ctcggatccc atcgcagcta ccgcggctag
     1261 ccaccatggc ttccaaggtg tacgaccccg agcaacgcaa acgcatgatc actgggcctc
     1321 agtggtgggc tcgctgcaag caaatgaacg tgctggactc cttcatcaac tactatgatt
     1381 ccgagaagca cgccgagaac gccgtgattt ttctgcatgg taacgctgcc tccagctacc
     1441 tgtggaggca cgtcgtgcct cacatcgagc ccgtggctag atgcatcatc cctgatctga
     1501 tcggaatggg taagtccggc aagagcggga atggctcata tcgcctcctg gatcactaca
     1561 agtacctcac cgcttggttc gagctgctga accttccaaa gaaaatcatc tttgtgggcc
     1621 acgactgggg ggcttgtctg gcctttcact actcctacga gcaccaagac aagatcaagg
     1681 ccatcgtcca tgctgagagt gtcgtggacg tgatcgagtc ctgggacgag tggcctgaca
     1741 tcgaggagga tatcgccctg atcaagagcg aagagggcga gaaaatggtg cttgagaata
     1801 acttcttcgt cgagaccatg ctcccaagca agatcatgcg gaaactggag cctgaggagt
     1861 tcgctgccta cctggagcca ttcaaggaga agggcgaggt tagacggcct accctctcct
     1921 ggcctcgcga gatccctctc gttaagggag gcaagcccga cgtcgtccag attgtccgca
     1981 actacaacgc ctaccttcgg gccagcgacg atctgcctaa gatgttcatc gagtccgacc
     2041 ctgggttctt ttccaacgct attgtcgagg gagctaagaa gttccctaac accgagttcg
     2101 tgaaggtgaa gggcctccac ttcagccagg aggacgctcc agatgaaatg ggtaagtaca
     2161 tcaagagctt cgtggagcgc gtgctgaaga acgagcagta attctaggcg atcgctcgag
     2221 cccgggaatt cgtttaaacc tagagcggcc gctggccgca ataaaatatc tttattttca
     2281 ttacatctgt gtgttggttt tttgtgtgag gatctaaatg agtcttcgga cctcgcgggg
     2341 gccgcttaag cggtggttag ggtttgtctg acgcgggggg agggggaagg aacgaaacac
     2401 tctcattcgg aggcggctcg gggtttggtc ttggtggcca cgggcacgca gaagagcgcc
     2461 gcgatcctct taagcacccc cccgccctcc gtggaggcgg gggtttggtc ggcgggtggt
     2521 aactggcggg ccgctgactc gggcgggtcg cgcgccccag agtgtgacct tttcggtctg
     2581 ctcgcagacc cccgggcggc gccgccgcgg cggcgacggg ctcgctgggt cctaggctcc
     2641 atggggaccg tatacgtgga caggctctgg agcatccgca cgactgcggt gatattaccg
     2701 gagaccttct gcgggacgag ccgggtcacg cggctgacgc ggagcgtccg ttgggcgaca
     2761 aacaccagga cggggcacag gtacactatc ttgtcacccg gaggcgcgag ggactgcagg
     2821 agcttcaggg agtggcgcag ctgcttcatc cccgtggccc gttgctcgcg tttgctggcg
     2881 gtgtccccgg aagaaatata tttgcatgtc tttagttcta tgatgacaca aaccccgccc
     2941 agcgtcttgt cattggcgaa ttcgaacacg cagatgcagt cggggcggcg cggtcccagg
     3001 tccacttcgc atattaaggt gacgcgtgtg gcctcgaaca ccgagcgacc ctgcagcgac
     3061 ccgcttaaaa gcttggcatt ccggtactgt tggtaaagcc accatggccg atgctaagaa
     3121 cattaagaag ggccctgctc ccttctaccc tctggaggat ggcaccgctg gcgagcagct
     3181 gcacaaggcc atgaagaggt atgccctggt gcctggcacc attgccttca ccgatgccca
     3241 cattgaggtg gacatcacct atgccgagta cttcgagatg tctgtgcgcc tggccgaggc
     3301 catgaagagg tacggcctga acaccaacca ccgcatcgtg gtgtgctctg agaactctct
     3361 gcagttcttc atgccagtgc tgggcgccct gttcatcgga gtggccgtgg cccctgctaa
     3421 cgacatttac aacgagcgcg agctgctgaa cagcatgggc atttctcagc ctaccgtggt
     3481 gttcgtgtct aagaagggcc tgcagaagat cctgaacgtg cagaagaagc tgcctatcat
     3541 ccagaagatc atcatcatgg actctaagac cgactaccag ggcttccaga gcatgtacac
     3601 attcgtgaca tctcatctgc ctcctggctt caacgagtac gacttcgtgc cagagtcttt
     3661 cgacagggac aaaaccattg ccctgatcat gaacagctct gggtctaccg gcctgcctaa
     3721 gggcgtggcc ctgcctcatc gcaccgcctg tgtgcgcttc tctcacgccc gcgaccctat
     3781 tttcggcaac cagatcatcc ccgacaccgc tattctgagc gtggtgccat tccaccacgg
     3841 cttcggcatg ttcaccaccc tgggctacct gatttgcggc tttcgggtgg tgctgatgta
     3901 ccgcttcgag gaggagctgt tcctgcgcag cctgcaagac tacaaaattc agtctgccct
     3961 gctggtgcca accctgttca gcttcttcgc taagagcacc ctgatcgaca agtacgacct
     4021 gtctaacctg cacgagattg cctctggcgg cgccccactg tctaaggagg tgggcgaagc
     4081 cgtggccaag cgctttcatc tgccaggcat ccgccagggc tacggcctga ccgagacaac
     4141 cagcgccatt ctgattaccc cagagggcga cgacaagcct ggcgccgtgg gcaaggtggt
     4201 gccattcttc gaggccaagg tggtggacct ggacaccggc aagaccctgg gagtgaacca
     4261 gcgcggcgag ctgtgtgtgc gcggccctat gattatgtcc ggctacgtga ataaccctga
     4321 ggccacaaac gccctgatcg acaaggacgg ctggctgcac tctggcgaca ttgcctactg
     4381 ggacgaggac gagcacttct tcatcgtgga ccgcctgaag tctctgatca agtacaaggg
     4441 ctaccaggtg gccccagccg agctggagtc tatcctgctg cagcacccta acattttcga
     4501 cgccggagtg gccggcctgc ccgacgacga tgccggcgag ctgcctgccg ccgtcgtcgt
     4561 gctggaacac ggcaagacca tgaccgagaa ggagatcgtg gactatgtgg ccagccaggt
     4621 gacaaccgcc aagaagctgc gcggcggagt ggtgttcgtg gacgaggtgc ccaagggcct
     4681 gaccggcaag ctggacgccc gcaagatccg cgagatcctg atcaaggcta agaaaggcgg
     4741 caagatcgcc gtgtaataat tctagagtcg gggcggccgg ccgcttcgag cagacatgat
     4801 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     4861 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     4921 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     4981 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatccag gtggcacttt
     5041 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5101 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     5161 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5221 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     5281 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5341 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5401 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5461 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5521 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5581 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5641 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5701 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5761 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5821 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5881 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5941 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6001 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     6061 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6121 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6181 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6241 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6301 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6361 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6421 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6481 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6541 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6601 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6661 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6721 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6781 cgccagcaac gcggcctttt tacggttcct g
//
