LOCUS       sequence-229531-        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-KIT3
FEATURES             Location/Qualifiers
     source          1..6811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1243..1252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1249..2184
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2243..2291
                     /note="synthetic polyadenylation signal"
     promoter        2300..3051
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      3081..3090
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3087..4739
                     /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             4046..4054
                     /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    4783..4904
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4794..4813)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4848..4867
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5037..5141
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5142..5210,5211..6002)
                     /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(5360..5379)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6173..6761
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6662..6681
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgctcacatg gctcgacaga tctgagggat gtcttattag tttatcagaa tgcataggga
       61 aagctcatct ttcttggagt aggtcttcca ataagtgttc aagccctggg tcttccctca
      121 ctacaataat aaacatacct gctctaccct cctcacaaag ctgttatcgt gagcatgaaa
      181 agacgagtcg ctgtggggaa gaactattaa cccctataca gatactgttg ttgctttccg
      241 ttcaaggatc caccatgaga ctgtgggtaa ggcactttct gggcagtgaa atcgggctta
      301 agagaatcat tgtactttta ttttgctcct gggaccggca agggcagtta agagccctag
      361 ccctaaaact gtcgtctgtt acagaggcag agaaatcatt actggggttg gcagcgttgg
      421 ggggaggggg gaaaacgtgt atgaaaacct gggctcccag agcaaacttc gccacgcgcc
      481 ctccatcgcc cctgtctcca cacaagtccc ggcaggggct tcttgggcgg aggccacgcc
      541 gaccaatagg aacccactgt gttcctacag gttacgaagc aggtggagaa attgagcaga
      601 aacaattagc gaaaccgggc tcagccttta ccgcggtgcc aggagctcct aacaggcctg
      661 gaggggaatg tcggggctca atttcctaac gctcccctcc ccatccccat gccacctcca
      721 cgagcagcgg cgtccagcct cctcccgccc gaacgtgctc gaggggcggg cagtcgacct
      781 ttattgtctg gggagcacct ggcaggtggc gggcccgtgc cctaacgtgt gcgtggtgcc
      841 cagcttcaca aagcgagcgg gcagcacctc cttggtccgg gaacgcctca gcctggccgt
      901 ccacatccca ggggtggaaa ggtggagaga gaaaggggct ccggagtcaa gagcggggag
      961 agagggcgcg cgccctcctc ctcccggcgg gcacagcccc ccggcattaa cacgtcgaaa
     1021 gagcaggggc cagacgccgc cgggaagaag cgagacccgg gcgggcgcga gggaggggag
     1081 acgagaagaa acgtgaccga cgcgcagagg gagggcgctg ggaggagggg ctgctgctcg
     1141 ccgctcgcgg ctctgggggc tcggctttgc cgcgctcgct gcacttgggc gagagctgga
     1201 acgtggacca gagctcggat cccatcgcag ctaccgcggc tagccaccat ggcttccaag
     1261 gtgtacgacc ccgagcaacg caaacgcatg atcactgggc ctcagtggtg ggctcgctgc
     1321 aagcaaatga acgtgctgga ctccttcatc aactactatg attccgagaa gcacgccgag
     1381 aacgccgtga tttttctgca tggtaacgct gcctccagct acctgtggag gcacgtcgtg
     1441 cctcacatcg agcccgtggc tagatgcatc atccctgatc tgatcggaat gggtaagtcc
     1501 ggcaagagcg ggaatggctc atatcgcctc ctggatcact acaagtacct caccgcttgg
     1561 ttcgagctgc tgaaccttcc aaagaaaatc atctttgtgg gccacgactg gggggcttgt
     1621 ctggcctttc actactccta cgagcaccaa gacaagatca aggccatcgt ccatgctgag
     1681 agtgtcgtgg acgtgatcga gtcctgggac gagtggcctg acatcgagga ggatatcgcc
     1741 ctgatcaaga gcgaagaggg cgagaaaatg gtgcttgaga ataacttctt cgtcgagacc
     1801 atgctcccaa gcaagatcat gcggaaactg gagcctgagg agttcgctgc ctacctggag
     1861 ccattcaagg agaagggcga ggttagacgg cctaccctct cctggcctcg cgagatccct
     1921 ctcgttaagg gaggcaagcc cgacgtcgtc cagattgtcc gcaactacaa cgcctacctt
     1981 cgggccagcg acgatctgcc taagatgttc atcgagtccg accctgggtt cttttccaac
     2041 gctattgtcg agggagctaa gaagttccct aacaccgagt tcgtgaaggt gaagggcctc
     2101 cacttcagcc aggaggacgc tccagatgaa atgggtaagt acatcaagag cttcgtggag
     2161 cgcgtgctga agaacgagca gtaattctag gcgatcgctc gagcccggga attcgtttaa
     2221 acctagagcg gccgctggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg
     2281 ttttttgtgt gaggatctaa atgagtcttc ggacctcgcg ggggccgctt aagcggtggt
     2341 tagggtttgt ctgacgcggg gggaggggga aggaacgaaa cactctcatt cggaggcggc
     2401 tcggggtttg gtcttggtgg ccacgggcac gcagaagagc gccgcgatcc tcttaagcac
     2461 ccccccgccc tccgtggagg cgggggtttg gtcggcgggt ggtaactggc gggccgctga
     2521 ctcgggcggg tcgcgcgccc cagagtgtga ccttttcggt ctgctcgcag acccccgggc
     2581 ggcgccgccg cggcggcgac gggctcgctg ggtcctaggc tccatgggga ccgtatacgt
     2641 ggacaggctc tggagcatcc gcacgactgc ggtgatatta ccggagacct tctgcgggac
     2701 gagccgggtc acgcggctga cgcggagcgt ccgttgggcg acaaacacca ggacggggca
     2761 caggtacact atcttgtcac ccggaggcgc gagggactgc aggagcttca gggagtggcg
     2821 cagctgcttc atccccgtgg cccgttgctc gcgtttgctg gcggtgtccc cggaagaaat
     2881 atatttgcat gtctttagtt ctatgatgac acaaaccccg cccagcgtct tgtcattggc
     2941 gaattcgaac acgcagatgc agtcggggcg gcgcggtccc aggtccactt cgcatattaa
     3001 ggtgacgcgt gtggcctcga acaccgagcg accctgcagc gacccgctta aaagcttggc
     3061 attccggtac tgttggtaaa gccaccatgg ccgatgctaa gaacattaag aagggccctg
     3121 ctcccttcta ccctctggag gatggcaccg ctggcgagca gctgcacaag gccatgaaga
     3181 ggtatgccct ggtgcctggc accattgcct tcaccgatgc ccacattgag gtggacatca
     3241 cctatgccga gtacttcgag atgtctgtgc gcctggccga ggccatgaag aggtacggcc
     3301 tgaacaccaa ccaccgcatc gtggtgtgct ctgagaactc tctgcagttc ttcatgccag
     3361 tgctgggcgc cctgttcatc ggagtggccg tggcccctgc taacgacatt tacaacgagc
     3421 gcgagctgct gaacagcatg ggcatttctc agcctaccgt ggtgttcgtg tctaagaagg
     3481 gcctgcagaa gatcctgaac gtgcagaaga agctgcctat catccagaag atcatcatca
     3541 tggactctaa gaccgactac cagggcttcc agagcatgta cacattcgtg acatctcatc
     3601 tgcctcctgg cttcaacgag tacgacttcg tgccagagtc tttcgacagg gacaaaacca
     3661 ttgccctgat catgaacagc tctgggtcta ccggcctgcc taagggcgtg gccctgcctc
     3721 atcgcaccgc ctgtgtgcgc ttctctcacg cccgcgaccc tattttcggc aaccagatca
     3781 tccccgacac cgctattctg agcgtggtgc cattccacca cggcttcggc atgttcacca
     3841 ccctgggcta cctgatttgc ggctttcggg tggtgctgat gtaccgcttc gaggaggagc
     3901 tgttcctgcg cagcctgcaa gactacaaaa ttcagtctgc cctgctggtg ccaaccctgt
     3961 tcagcttctt cgctaagagc accctgatcg acaagtacga cctgtctaac ctgcacgaga
     4021 ttgcctctgg cggcgcccca ctgtctaagg aggtgggcga agccgtggcc aagcgctttc
     4081 atctgccagg catccgccag ggctacggcc tgaccgagac aaccagcgcc attctgatta
     4141 ccccagaggg cgacgacaag cctggcgccg tgggcaaggt ggtgccattc ttcgaggcca
     4201 aggtggtgga cctggacacc ggcaagaccc tgggagtgaa ccagcgcggc gagctgtgtg
     4261 tgcgcggccc tatgattatg tccggctacg tgaataaccc tgaggccaca aacgccctga
     4321 tcgacaagga cggctggctg cactctggcg acattgccta ctgggacgag gacgagcact
     4381 tcttcatcgt ggaccgcctg aagtctctga tcaagtacaa gggctaccag gtggccccag
     4441 ccgagctgga gtctatcctg ctgcagcacc ctaacatttt cgacgccgga gtggccggcc
     4501 tgcccgacga cgatgccggc gagctgcctg ccgccgtcgt cgtgctggaa cacggcaaga
     4561 ccatgaccga gaaggagatc gtggactatg tggccagcca ggtgacaacc gccaagaagc
     4621 tgcgcggcgg agtggtgttc gtggacgagg tgcccaaggg cctgaccggc aagctggacg
     4681 cccgcaagat ccgcgagatc ctgatcaagg ctaagaaagg cggcaagatc gccgtgtaat
     4741 aattctagag tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt
     4801 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     4861 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     4921 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     4981 tctacaaatg tggtaaaatc gataaggatc caggtggcac ttttcgggga aatgtgcgcg
     5041 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5101 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5161 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5221 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5281 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5341 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5401 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5461 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5521 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5581 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5641 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5701 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5761 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5821 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5881 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5941 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6001 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6061 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6121 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6181 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6241 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6301 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6361 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6421 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6481 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6541 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6601 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6661 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6721 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6781 ttttacggtt cctggccttt tgctggcctt t
//
