LOCUS       sequence-229534-        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-KIT4
FEATURES             Location/Qualifiers
     source          1..6811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1283..1292
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1289..2224
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2283..2331
                     /note="synthetic polyadenylation signal"
     promoter        2340..3091
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      3121..3130
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3127..4779
                     /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             4086..4094
                     /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    4823..4944
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4834..4853)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4888..4907
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5077..5181
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5182..5250,5251..6042)
                     /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(5400..5419)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6213..6801
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6702..6721
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg gctcgacaga
       61 tctgagggat gtcttattag tttatcagaa tgcataggga aagctcatct ttcttggagt
      121 aggtcttcca ataagtgttc aagccctggg tcttccctca ctacaataat aaacatacct
      181 gctctaccct cctcacaaag ctgttatcgt gagcatgaaa agacgagtcg ctgtggggaa
      241 gaactattaa cccctataca gatactgttg ttgctttccg ttcaaggatc caccatgaga
      301 ctgtgggtaa ggcactttct gggcagtgaa atcgggctta agagaatcat tgtactttta
      361 ttttgctcct gggaccggca agggcagtta agagccctag ccctaaaact gtcgtctgtt
      421 acagaggcag agaaatcatt actggggttg gcagcgttgg ggggaggggg gaaaacgtgt
      481 atgaaaacct gggctcccag agcaaacttc gccacgcgcc ctccatcgcc cctgtctcca
      541 cacaagtccc ggcaggggct tcttgggcgg aggccacgcc gaccaatagg aacccactgt
      601 gttcctacag gttacgaagc aggtggagaa attgagcaga aacaattagc gaaaccgggc
      661 tcagccttta ccgcggtgcc aggagctcct aacaggcctg gaggggaatg tcggggctca
      721 atttcctaac gctcccctcc ccatccccat gccacctcca cgagcagcgg cgtccagcct
      781 cctcccgccc gaacgtgctc gaggggcggg cagtcgacct ttattgtctg gggagcacct
      841 ggcaggtggc gggcccgtgc cctaacgtgt gcgtggtgcc cagcttcaca aagcgagcgg
      901 gcagcacctc cttggtccgg gaacgcctca gcctggccgt ccacatccca ggggtggaaa
      961 ggtggagaga gaaaggggct ccggagtcaa gagcggggag agagggcgcg cgccctcctc
     1021 ctcccggcgg gcacagcccc ccggcattaa cacgtcgaaa gagcaggggc cagacgccgc
     1081 cgggaagaag cgagacccgg gcgggcgcga gggaggggag gcgaggaggg gcgtggccgg
     1141 cgcgcagaaa gagaacgcta agaggaggaa ctgctgctcg ccgctcgcgg ctctgggggc
     1201 tcggctttgc cgcgctcgct gcacttgggc gagagctgga acgtggacca gagctcggat
     1261 cccatcgcag ctaccgcggc tagccaccat ggcttccaag gtgtacgacc ccgagcaacg
     1321 caaacgcatg atcactgggc ctcagtggtg ggctcgctgc aagcaaatga acgtgctgga
     1381 ctccttcatc aactactatg attccgagaa gcacgccgag aacgccgtga tttttctgca
     1441 tggtaacgct gcctccagct acctgtggag gcacgtcgtg cctcacatcg agcccgtggc
     1501 tagatgcatc atccctgatc tgatcggaat gggtaagtcc ggcaagagcg ggaatggctc
     1561 atatcgcctc ctggatcact acaagtacct caccgcttgg ttcgagctgc tgaaccttcc
     1621 aaagaaaatc atctttgtgg gccacgactg gggggcttgt ctggcctttc actactccta
     1681 cgagcaccaa gacaagatca aggccatcgt ccatgctgag agtgtcgtgg acgtgatcga
     1741 gtcctgggac gagtggcctg acatcgagga ggatatcgcc ctgatcaaga gcgaagaggg
     1801 cgagaaaatg gtgcttgaga ataacttctt cgtcgagacc atgctcccaa gcaagatcat
     1861 gcggaaactg gagcctgagg agttcgctgc ctacctggag ccattcaagg agaagggcga
     1921 ggttagacgg cctaccctct cctggcctcg cgagatccct ctcgttaagg gaggcaagcc
     1981 cgacgtcgtc cagattgtcc gcaactacaa cgcctacctt cgggccagcg acgatctgcc
     2041 taagatgttc atcgagtccg accctgggtt cttttccaac gctattgtcg agggagctaa
     2101 gaagttccct aacaccgagt tcgtgaaggt gaagggcctc cacttcagcc aggaggacgc
     2161 tccagatgaa atgggtaagt acatcaagag cttcgtggag cgcgtgctga agaacgagca
     2221 gtaattctag gcgatcgctc gagcccggga attcgtttaa acctagagcg gccgctggcc
     2281 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaggatctaa
     2341 atgagtcttc ggacctcgcg ggggccgctt aagcggtggt tagggtttgt ctgacgcggg
     2401 gggaggggga aggaacgaaa cactctcatt cggaggcggc tcggggtttg gtcttggtgg
     2461 ccacgggcac gcagaagagc gccgcgatcc tcttaagcac ccccccgccc tccgtggagg
     2521 cgggggtttg gtcggcgggt ggtaactggc gggccgctga ctcgggcggg tcgcgcgccc
     2581 cagagtgtga ccttttcggt ctgctcgcag acccccgggc ggcgccgccg cggcggcgac
     2641 gggctcgctg ggtcctaggc tccatgggga ccgtatacgt ggacaggctc tggagcatcc
     2701 gcacgactgc ggtgatatta ccggagacct tctgcgggac gagccgggtc acgcggctga
     2761 cgcggagcgt ccgttgggcg acaaacacca ggacggggca caggtacact atcttgtcac
     2821 ccggaggcgc gagggactgc aggagcttca gggagtggcg cagctgcttc atccccgtgg
     2881 cccgttgctc gcgtttgctg gcggtgtccc cggaagaaat atatttgcat gtctttagtt
     2941 ctatgatgac acaaaccccg cccagcgtct tgtcattggc gaattcgaac acgcagatgc
     3001 agtcggggcg gcgcggtccc aggtccactt cgcatattaa ggtgacgcgt gtggcctcga
     3061 acaccgagcg accctgcagc gacccgctta aaagcttggc attccggtac tgttggtaaa
     3121 gccaccatgg ccgatgctaa gaacattaag aagggccctg ctcccttcta ccctctggag
     3181 gatggcaccg ctggcgagca gctgcacaag gccatgaaga ggtatgccct ggtgcctggc
     3241 accattgcct tcaccgatgc ccacattgag gtggacatca cctatgccga gtacttcgag
     3301 atgtctgtgc gcctggccga ggccatgaag aggtacggcc tgaacaccaa ccaccgcatc
     3361 gtggtgtgct ctgagaactc tctgcagttc ttcatgccag tgctgggcgc cctgttcatc
     3421 ggagtggccg tggcccctgc taacgacatt tacaacgagc gcgagctgct gaacagcatg
     3481 ggcatttctc agcctaccgt ggtgttcgtg tctaagaagg gcctgcagaa gatcctgaac
     3541 gtgcagaaga agctgcctat catccagaag atcatcatca tggactctaa gaccgactac
     3601 cagggcttcc agagcatgta cacattcgtg acatctcatc tgcctcctgg cttcaacgag
     3661 tacgacttcg tgccagagtc tttcgacagg gacaaaacca ttgccctgat catgaacagc
     3721 tctgggtcta ccggcctgcc taagggcgtg gccctgcctc atcgcaccgc ctgtgtgcgc
     3781 ttctctcacg cccgcgaccc tattttcggc aaccagatca tccccgacac cgctattctg
     3841 agcgtggtgc cattccacca cggcttcggc atgttcacca ccctgggcta cctgatttgc
     3901 ggctttcggg tggtgctgat gtaccgcttc gaggaggagc tgttcctgcg cagcctgcaa
     3961 gactacaaaa ttcagtctgc cctgctggtg ccaaccctgt tcagcttctt cgctaagagc
     4021 accctgatcg acaagtacga cctgtctaac ctgcacgaga ttgcctctgg cggcgcccca
     4081 ctgtctaagg aggtgggcga agccgtggcc aagcgctttc atctgccagg catccgccag
     4141 ggctacggcc tgaccgagac aaccagcgcc attctgatta ccccagaggg cgacgacaag
     4201 cctggcgccg tgggcaaggt ggtgccattc ttcgaggcca aggtggtgga cctggacacc
     4261 ggcaagaccc tgggagtgaa ccagcgcggc gagctgtgtg tgcgcggccc tatgattatg
     4321 tccggctacg tgaataaccc tgaggccaca aacgccctga tcgacaagga cggctggctg
     4381 cactctggcg acattgccta ctgggacgag gacgagcact tcttcatcgt ggaccgcctg
     4441 aagtctctga tcaagtacaa gggctaccag gtggccccag ccgagctgga gtctatcctg
     4501 ctgcagcacc ctaacatttt cgacgccgga gtggccggcc tgcccgacga cgatgccggc
     4561 gagctgcctg ccgccgtcgt cgtgctggaa cacggcaaga ccatgaccga gaaggagatc
     4621 gtggactatg tggccagcca ggtgacaacc gccaagaagc tgcgcggcgg agtggtgttc
     4681 gtggacgagg tgcccaaggg cctgaccggc aagctggacg cccgcaagat ccgcgagatc
     4741 ctgatcaagg ctaagaaagg cggcaagatc gccgtgtaat aattctagag tcggggcggc
     4801 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     4861 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     4921 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     4981 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
     5041 gataaggatc caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5101 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5161 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5221 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5281 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5341 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5401 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5461 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5521 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5581 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5641 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5701 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5761 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5821 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5881 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5941 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6001 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6061 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6121 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6181 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6241 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6301 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6361 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6421 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6481 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6541 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6601 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6661 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6721 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6781 ggggggcgga gcctatggaa aaacgccagc a
//
