LOCUS       sequence-451021-        6412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  0.5 kb from mouse Ccnd2 promoter in front of firefly luciferase 
            reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6412)
  AUTHORS   Fraire CR, Desai K, Obalapuram UA, Mendyka LK, Rajaram V, Sebastian 
            T, Wang Y, Onel K, Lee J, Chen KS
  TITLE     An imbalance between proliferation and differentiation underlies the
            development of microRNA-defective pineoblastoma.
  JOURNAL   bioRxiv [Preprint]. 2024 Apr 30:2024.04.23.590638. doi: 
            10.1101/2024.04.23.590638.
  PUBMED    38712047
REFERENCE   2  (bases 1 to 6412)
  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"; journalName: "bioRxiv 
            [Preprint]. 2024 Apr 30:2024.04.23.590638. doi: 
            10.1101/2024.04.23.590638."
COMMENT     Sequence Label: pGL4.26-Ccnd2PromoterRegion2(0.5kb)
FEATURES             Location/Qualifiers
     source          1..6412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        607..638
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      665..674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             671..2323
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    2367..2488
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2378..2397)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2432..2451
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2682..3039
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2682..2873
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2705..2725)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2890..3025
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2952..2971
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3070..4107
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     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    4131..4179
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4335..4352)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4506..5094)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4586..4605)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5294..6085,6086..6154))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    6259..6307
                     /note="synthetic polyadenylation signal"
     misc_feature    6321..6412
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6361..6380
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacca atctgcccgg taacaatcag cgcgcagtag caggagcccc
       61 agagctattg gctatgcaaa tagagggagg ggagacggcg ccccaaactc atactcaccc
      121 ttttaaagtg atatcccctc cccccgccca ccccttccgc cccaccctcg tttaaagggg
      181 ctggcccagg gctggagtta atcgcttgca cctcggcttt attgcctccc ctcccgggcc
      241 tttccgagag cccacgggtg gtgctcccac agcgtgccac cccctccccc ccccagcctc
      301 agcctccgct agccttaagc gtccctgatc atcccccctt cttaaggaag gagggactca
      361 gtgccccccc ccgtgtgccc agagaggtgc aacccacgcc atgctatctg cacgtgccag
      421 cgtggccacc atgcgggggc cagtggttcc tcgccccttc ctccctcctt taatatagtt
      481 aaaggcaggg tgggagcagg aaggaggtga ggaaacgcca acgtgtcccc ctctaccaac
      541 agccttgacc acaggatgct ttctcgagga tatcaagatc tggcctcggc ggccaagctt
      601 agacactaga gggtatataa tggaagctcg acttccagct tggcaatccg gtactgttgg
      661 taaagccacc atggaagatg ccaaaaacat taagaagggc ccagcgccat tctacccact
      721 cgaagacggg accgccggcg agcagctgca caaagccatg aagcgctacg ccctggtgcc
      781 cggcaccatc gcctttaccg acgcacatat cgaggtggac attacctacg ccgagtactt
      841 cgagatgagc gttcggctgg cagaagctat gaagcgctat gggctgaata caaaccatcg
      901 gatcgtggtg tgcagcgaga atagcttgca gttcttcatg cccgtgttgg gtgccctgtt
      961 catcggtgtg gctgtggccc cagctaacga catctacaac gagcgcgagc tgctgaacag
     1021 catgggcatc agccagccca ccgtcgtatt cgtgagcaag aaagggctgc aaaagatcct
     1081 caacgtgcaa aagaagctac cgatcataca aaagatcatc atcatggata gcaagaccga
     1141 ctaccagggc ttccaaagca tgtacacctt cgtgacttcc catttgccac ccggcttcaa
     1201 cgagtacgac ttcgtgcccg agagcttcga ccgggacaaa accatcgccc tgatcatgaa
     1261 cagtagtggc agtaccggat tgcccaaggg cgtagcccta ccgcaccgca ccgcttgtgt
     1321 ccgattcagt catgcccgcg accccatctt cggcaaccag atcatccccg acaccgctat
     1381 cctcagcgtg gtgccatttc accacggctt cggcatgttc accacgctgg gctacttgat
     1441 ctgcggcttt cgggtcgtgc tcatgtaccg cttcgaggag gagctattct tgcgcagctt
     1501 gcaagactat aagattcaat ctgccctgct ggtgcccaca ctatttagct tcttcgctaa
     1561 gagcactctc atcgacaagt acgacctaag caacttgcac gagatcgcca gcggcggggc
     1621 gccgctcagc aaggaggtag gtgaggccgt ggccaaacgc ttccacctac caggcatccg
     1681 ccagggctac ggcctgacag aaacaaccag cgccattctg atcacccccg aaggggacga
     1741 caagcctggc gcagtaggca aggtggtgcc cttcttcgag gctaaggtgg tggacttgga
     1801 caccggtaag acactgggtg tgaaccagcg cggcgagctg tgcgtccgtg gccccatgat
     1861 catgagcggc tacgttaaca accccgaggc tacaaacgct ctcatcgaca aggacggctg
     1921 gctgcacagc ggcgacatcg cctactggga cgaggacgag cacttcttca tcgtggaccg
     1981 gctgaagagc ctgatcaaat acaagggcta ccaggtagcc ccagccgaac tggagagcat
     2041 cctgctgcaa caccccaaca tcttcgacgc cggggtcgcc ggcctgcccg acgacgatgc
     2101 cggcgagctg cccgccgcag tcgtcgtgct ggaacacggt aaaaccatga ccgagaagga
     2161 gatcgtggac tatgtggcca gccaggttac aaccgccaag aagctgcgcg gtggtgttgt
     2221 gttcgtggac gaggtgccta aaggactgac cggcaagttg gacgcccgca agatccgcga
     2281 gattctcatt aaggccaaga agggcggcaa gatcgccgtg taataattct agagtcgggg
     2341 cggccggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     2401 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2461 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     2521 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa
     2581 aatcgataag gatccgtttg cgtattgggc gctcttccgc tgatctgcgc agcaccatgg
     2641 cctgaaataa cctctgaaag aggaacttgg ttagctacct tctgaggcgg aaagaaccag
     2701 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2761 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2821 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2881 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2941 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3001 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcgattc ttctgacact
     3061 agcgccacca tgaagaagcc cgaactcacc gctaccagcg ttgaaaaatt tctcatcgag
     3121 aagttcgaca gtgtgagcga cctgatgcag ttgtcggagg gcgaagagag ccgagccttc
     3181 agcttcgatg tcggcggacg cggctatgta ctgcgggtga atagctgcgc tgatggcttc
     3241 tacaaagacc gctacgtgta ccgccacttc gccagcgctg cactacccat ccccgaagtg
     3301 ttggacatcg gcgagttcag cgagagcctg acatactgca tcagtagacg cgcccaaggc
     3361 gttactctcc aagacctccc cgaaacagag ctgcctgctg tgttacagcc tgtcgccgaa
     3421 gctatggatg ctattgccgc cgccgacctc agtcaaacca gcggcttcgg cccattcggg
     3481 ccccaaggca tcggccagta cacaacctgg cgggatttca tttgcgccat tgctgatccc
     3541 catgtctacc actggcagac cgtgatggac gacaccgtgt ccgccagcgt agctcaagcc
     3601 ctggacgaac tgatgctgtg ggccgaagac tgtcccgagg tgcgccacct cgtccatgcc
     3661 gacttcggca gcaacaacgt cctgaccgac aacggccgca tcaccgccgt aatcgactgg
     3721 tccgaagcta tgttcgggga cagtcagtac gaggtggcca acatcttctt ctggcggccc
     3781 tggctggctt gcatggagca gcagactcgc tacttcgagc gccggcatcc cgagctggcc
     3841 ggcagccctc gtctgcgagc ctacatgctg cgcatcggcc tggatcagct ctaccagagc
     3901 ctcgtggacg gcaacttcga cgatgctgcc tgggctcaag gccgctgcga tgccatcgtc
     3961 cgcagcgggg ccggcaccgt cggtcgcaca caaatcgctc gccggagcgc agccgtatgg
     4021 accgacggct gcgtcgaggt gctggccgac agcggcaacc gccggcccag tacacgaccg
     4081 cgcgctaagg aggtaggtcg agtttaaact ctagaaccgg tcatggccgc aataaaatat
     4141 ctttattttc attacatctg tgtgttggtt ttttgtgtgt tcgaactaga tgctgtcgac
     4201 cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta
     4261 tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag
     4321 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4381 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4441 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4501 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4561 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4621 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4681 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4741 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4801 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4861 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4921 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4981 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5041 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5101 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5161 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5221 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagcggccg caaatgctaa
     5281 accactgcag tggttaccag tgcttgatca gtgaggcacc gatctcagcg atctgcctat
     5341 ttcgttcgtc catagtggcc tgactccccg tcgtgtagat cactacgatt cgtgagggct
     5401 taccatcagg ccccagcgca gcaatgatgc cgcgagagcc gcgttcaccg gcccccgatt
     5461 tgtcagcaat gaaccagcca gcagggaggg ccgagcgaag aagtggtcct gctactttgt
     5521 ccgcctccat ccagtctatg agctgctgtc gtgatgctag agtaagaagt tcgccagtga
     5581 gtagtttccg aagagttgtg gccattgcta ctggcatcgt ggtatcacgc tcgtcgttcg
     5641 gtatggcttc gttcaactct ggttcccagc ggtcaagccg ggtcacatga tcacccatat
     5701 tatgaagaaa tgcagtcagc tccttagggc ctccgatcgt tgtcagaagt aagttggccg
     5761 cggtgttgtc gctcatggta atggcagcac tacacaattc tcttaccgtc atgccatccg
     5821 taagatgctt ttccgtgacc ggcgagtact caaccaagtc gttttgtgag tagtgtatac
     5881 ggcgaccaag ctgctcttgc ccggcgtcta tacgggacaa caccgcgcca catagcagta
     5941 ctttgaaagt gctcatcatc gggaatcgtt cttcggggcg gaaagactca aggatcttgc
     6001 cgctattgag atccagttcg atatagccca ctcttgcacc cagttgatct tcagcatctt
     6061 ttactttcac cagcgtttcg gggtgtgcaa aaacaggcaa gcaaaatgcc gcaaagaagg
     6121 gaatgagtgc gacacgaaaa tgttggatgc tcatactcgt cctttttcaa tattattgaa
     6181 gcatttatca gggttactag tacgtctctc aaggataagt aagtaatatt aaggtacggg
     6241 aggtattgga caggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt
     6301 ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa
     6361 ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct ct
//
