LOCUS       sequence-229526-        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-KIT1
FEATURES             Location/Qualifiers
     source          1..6811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1273..1282
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1279..2214
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2273..2321
                     /note="synthetic polyadenylation signal"
     promoter        2330..3081
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      3111..3120
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3117..4769
                     /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             4076..4084
                     /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    4813..4934
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4824..4843)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4878..4897
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5067..5171
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5172..5240,5241..6032)
                     /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(5390..5409)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6203..6791
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6692..6711
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttacggttc ctggcctttt gctggccttt tgctcacatg gctcgacaga tctgagggat
       61 gtcttattag tttatcagaa tgcataggga aagctcatct ttcttggagt aggtcttcca
      121 ataagtgttc aagccctggg tcttccctca ctacaataat aaacatacct gctctaccct
      181 cctcacaaag ctgttatcgt gagcatgaaa agacgagtcg ctgtggggaa gaactattaa
      241 cccctataca gatactgttg ttgctttccg ttcaaggatc caccatgaga ctgtgggtaa
      301 ggcactttct gggcagtgaa atcgggctta agagaatcat tgtactttta ttttgctcct
      361 gggaccggca agggcagtta agagccctag ccctaaaact gtcgtctgtt acagaggcag
      421 agaaatcatt actggggttg gcagcgttgg ggggaggggg gaaaacgtgt atgaaaacct
      481 gggctcccag agcaaacttc gccacgcgcc ctccatcgcc cctgtctcca cacaagtccc
      541 ggcaggggct tcttgggcgg aggccacgcc gaccaatagg aacccactgt gttcctacag
      601 gttacgaagc aggtggagaa attgagcaga aacaattagc gaaaccgggc tcagccttta
      661 ccgcggtgcc aggagctcct aacaggcctg gaggggaatg tcggggctca atttcctaac
      721 gctcccctcc ccatccccat gccacctcca cgagcagcgg cgtccagcct cctcccgccc
      781 gaacgtgctc gaggggcggg cagtcgacct ttattgtctg gggagcacct ggcaggtggc
      841 gggcccgtgc cctaacgtgt gcgtggtgcc cagcttcaca aagcgagcgg gcagcacctc
      901 cttggtccgg gaacgcctca gcctggccgt ccacatccca ggggtggaaa ggtggagaga
      961 gaaaggggct ccggagtcaa gagcggggag agagggcgcg cgccctcctc ctcccggcgg
     1021 gcacagcccc ccggcattaa cacgtcgaaa gagcaggggc cagacgccgc cgggaagaag
     1081 cgagacccgg gcgggcgcga gggaggggag gcgaggaggg gcgtggccgg cgcgcagagg
     1141 gagggcgctg ggaggagggg ctgctgctcg ccgctcgcgg ctctgggggc tcggctttgc
     1201 cgcgctcgct gcacttgggc gagagctgga acgtggacca gagctcggat cccatcgcag
     1261 ctaccgcggc tagccaccat ggcttccaag gtgtacgacc ccgagcaacg caaacgcatg
     1321 atcactgggc ctcagtggtg ggctcgctgc aagcaaatga acgtgctgga ctccttcatc
     1381 aactactatg attccgagaa gcacgccgag aacgccgtga tttttctgca tggtaacgct
     1441 gcctccagct acctgtggag gcacgtcgtg cctcacatcg agcccgtggc tagatgcatc
     1501 atccctgatc tgatcggaat gggtaagtcc ggcaagagcg ggaatggctc atatcgcctc
     1561 ctggatcact acaagtacct caccgcttgg ttcgagctgc tgaaccttcc aaagaaaatc
     1621 atctttgtgg gccacgactg gggggcttgt ctggcctttc actactccta cgagcaccaa
     1681 gacaagatca aggccatcgt ccatgctgag agtgtcgtgg acgtgatcga gtcctgggac
     1741 gagtggcctg acatcgagga ggatatcgcc ctgatcaaga gcgaagaggg cgagaaaatg
     1801 gtgcttgaga ataacttctt cgtcgagacc atgctcccaa gcaagatcat gcggaaactg
     1861 gagcctgagg agttcgctgc ctacctggag ccattcaagg agaagggcga ggttagacgg
     1921 cctaccctct cctggcctcg cgagatccct ctcgttaagg gaggcaagcc cgacgtcgtc
     1981 cagattgtcc gcaactacaa cgcctacctt cgggccagcg acgatctgcc taagatgttc
     2041 atcgagtccg accctgggtt cttttccaac gctattgtcg agggagctaa gaagttccct
     2101 aacaccgagt tcgtgaaggt gaagggcctc cacttcagcc aggaggacgc tccagatgaa
     2161 atgggtaagt acatcaagag cttcgtggag cgcgtgctga agaacgagca gtaattctag
     2221 gcgatcgctc gagcccggga attcgtttaa acctagagcg gccgctggcc gcaataaaat
     2281 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaggatctaa atgagtcttc
     2341 ggacctcgcg ggggccgctt aagcggtggt tagggtttgt ctgacgcggg gggaggggga
     2401 aggaacgaaa cactctcatt cggaggcggc tcggggtttg gtcttggtgg ccacgggcac
     2461 gcagaagagc gccgcgatcc tcttaagcac ccccccgccc tccgtggagg cgggggtttg
     2521 gtcggcgggt ggtaactggc gggccgctga ctcgggcggg tcgcgcgccc cagagtgtga
     2581 ccttttcggt ctgctcgcag acccccgggc ggcgccgccg cggcggcgac gggctcgctg
     2641 ggtcctaggc tccatgggga ccgtatacgt ggacaggctc tggagcatcc gcacgactgc
     2701 ggtgatatta ccggagacct tctgcgggac gagccgggtc acgcggctga cgcggagcgt
     2761 ccgttgggcg acaaacacca ggacggggca caggtacact atcttgtcac ccggaggcgc
     2821 gagggactgc aggagcttca gggagtggcg cagctgcttc atccccgtgg cccgttgctc
     2881 gcgtttgctg gcggtgtccc cggaagaaat atatttgcat gtctttagtt ctatgatgac
     2941 acaaaccccg cccagcgtct tgtcattggc gaattcgaac acgcagatgc agtcggggcg
     3001 gcgcggtccc aggtccactt cgcatattaa ggtgacgcgt gtggcctcga acaccgagcg
     3061 accctgcagc gacccgctta aaagcttggc attccggtac tgttggtaaa gccaccatgg
     3121 ccgatgctaa gaacattaag aagggccctg ctcccttcta ccctctggag gatggcaccg
     3181 ctggcgagca gctgcacaag gccatgaaga ggtatgccct ggtgcctggc accattgcct
     3241 tcaccgatgc ccacattgag gtggacatca cctatgccga gtacttcgag atgtctgtgc
     3301 gcctggccga ggccatgaag aggtacggcc tgaacaccaa ccaccgcatc gtggtgtgct
     3361 ctgagaactc tctgcagttc ttcatgccag tgctgggcgc cctgttcatc ggagtggccg
     3421 tggcccctgc taacgacatt tacaacgagc gcgagctgct gaacagcatg ggcatttctc
     3481 agcctaccgt ggtgttcgtg tctaagaagg gcctgcagaa gatcctgaac gtgcagaaga
     3541 agctgcctat catccagaag atcatcatca tggactctaa gaccgactac cagggcttcc
     3601 agagcatgta cacattcgtg acatctcatc tgcctcctgg cttcaacgag tacgacttcg
     3661 tgccagagtc tttcgacagg gacaaaacca ttgccctgat catgaacagc tctgggtcta
     3721 ccggcctgcc taagggcgtg gccctgcctc atcgcaccgc ctgtgtgcgc ttctctcacg
     3781 cccgcgaccc tattttcggc aaccagatca tccccgacac cgctattctg agcgtggtgc
     3841 cattccacca cggcttcggc atgttcacca ccctgggcta cctgatttgc ggctttcggg
     3901 tggtgctgat gtaccgcttc gaggaggagc tgttcctgcg cagcctgcaa gactacaaaa
     3961 ttcagtctgc cctgctggtg ccaaccctgt tcagcttctt cgctaagagc accctgatcg
     4021 acaagtacga cctgtctaac ctgcacgaga ttgcctctgg cggcgcccca ctgtctaagg
     4081 aggtgggcga agccgtggcc aagcgctttc atctgccagg catccgccag ggctacggcc
     4141 tgaccgagac aaccagcgcc attctgatta ccccagaggg cgacgacaag cctggcgccg
     4201 tgggcaaggt ggtgccattc ttcgaggcca aggtggtgga cctggacacc ggcaagaccc
     4261 tgggagtgaa ccagcgcggc gagctgtgtg tgcgcggccc tatgattatg tccggctacg
     4321 tgaataaccc tgaggccaca aacgccctga tcgacaagga cggctggctg cactctggcg
     4381 acattgccta ctgggacgag gacgagcact tcttcatcgt ggaccgcctg aagtctctga
     4441 tcaagtacaa gggctaccag gtggccccag ccgagctgga gtctatcctg ctgcagcacc
     4501 ctaacatttt cgacgccgga gtggccggcc tgcccgacga cgatgccggc gagctgcctg
     4561 ccgccgtcgt cgtgctggaa cacggcaaga ccatgaccga gaaggagatc gtggactatg
     4621 tggccagcca ggtgacaacc gccaagaagc tgcgcggcgg agtggtgttc gtggacgagg
     4681 tgcccaaggg cctgaccggc aagctggacg cccgcaagat ccgcgagatc ctgatcaagg
     4741 ctaagaaagg cggcaagatc gccgtgtaat aattctagag tcggggcggc cggccgcttc
     4801 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     4861 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     4921 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     4981 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc
     5041 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5101 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5161 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5221 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5281 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5341 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5401 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5461 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5521 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5581 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5641 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5701 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5761 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5821 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5881 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5941 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6001 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6061 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6121 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6181 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6241 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6301 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6361 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6421 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6481 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6541 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6601 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6661 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6721 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6781 gcctatggaa aaacgccagc aacgcggcct t
//
