LOCUS       sequence-451756-        7051 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  1.2 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 7051)
  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 7051)
  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-Ccnd2PromoterRegion1(1.2kb)
FEATURES             Location/Qualifiers
     source          1..7051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1246..1277
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      1304..1313
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1310..2962
                     /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    3006..3127
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3017..3036)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3071..3090
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3321..3678
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3321..3512
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3344..3364)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3529..3664
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3591..3610
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3709..4746
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4725..4733
                     /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    4770..4818
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4974..4991)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5145..5733)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5225..5244)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5933..6724,6725..6793))
                     /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    6898..6946
                     /note="synthetic polyadenylation signal"
     misc_feature    6960..7051
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     7000..7019
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct tttgatgtag cggagaaggc ttcctacttt aaaaaaaaaa
       61 agtaatctgc ccggtaacaa tcagcgcgca gtagcaggag ccccagagct attggctatg
      121 caaatagagg gaggggagac ggcgccccaa actcatactc acccttttaa agtgatatcc
      181 cctccccccg cccacccctt ccgccccacc ctcgtttaaa ggggctggcc cagggctgga
      241 gttaatcgct tgcacctcgg ctttattgcc tcccctcccg ggcctttccg agagcccacg
      301 ggtggtgctc ccacagcgtg ccaccccctc ccccccccag cctcagcctc cgctagcctt
      361 aagcgtccct gatcatcccc ccttcttaag gaaggaggga ctcagtgccc ccccccgtgt
      421 gcccagagag gtgcaaccca cgccatgcta tctgcacgtg ccagcgtggc caccatgcgg
      481 gggccagtgg ttcctcgccc cttcctccct cctttaatat agttaaaggc agggtgggag
      541 caggaaggag gtgaggaaac gccaacgtgt ccccctctac caacagcctt gaccacagga
      601 tgctttagag cacatgtcag agccgacccc aaaatcctgg cggaaggatg aggcgcgaga
      661 ccccacgggg tcggggattg aggggtgcag gactcaggag cccccgggga aaggaagggt
      721 ggccttgctg gcgcagggag agccaaacct aaaccctccc tctccccctc cctggcccct
      781 cccgagccat ttcctagaaa gctgtatcaa tgtggccacg ctccgcgcag acacctaggg
      841 cggcttgtca gcagatgcag gggcgaggaa gcgggttttt cctgcgtggc cgaggcccgc
      901 ggaggaaccg ctgctagccc tgcccccggt ctggggccgc tctgagaggc tggccgcgtt
      961 gcccaatggc gcccccaaga acagaaaggt ttctgcagga gggtcatatt ctactaggct
     1021 taagaagcac ccctttctcc aacatccacc ccttcctcct cttcctctcc acgcacgtgg
     1081 ctcggggcgg acaacagctt gaaagttatc aggagtctaa gcttgagggg cataaccttt
     1141 atccctggtt tggcgaggtt gcaattttct caaataagcc tttccttgtt cctcatttag
     1201 gctcgaggat atcaagatct ggcctcggcg gccaagctta gacactagag ggtatataat
     1261 ggaagctcga cttccagctt ggcaatccgg tactgttggt aaagccacca tggaagatgc
     1321 caaaaacatt aagaagggcc cagcgccatt ctacccactc gaagacggga ccgccggcga
     1381 gcagctgcac aaagccatga agcgctacgc cctggtgccc ggcaccatcg cctttaccga
     1441 cgcacatatc gaggtggaca ttacctacgc cgagtacttc gagatgagcg ttcggctggc
     1501 agaagctatg aagcgctatg ggctgaatac aaaccatcgg atcgtggtgt gcagcgagaa
     1561 tagcttgcag ttcttcatgc ccgtgttggg tgccctgttc atcggtgtgg ctgtggcccc
     1621 agctaacgac atctacaacg agcgcgagct gctgaacagc atgggcatca gccagcccac
     1681 cgtcgtattc gtgagcaaga aagggctgca aaagatcctc aacgtgcaaa agaagctacc
     1741 gatcatacaa aagatcatca tcatggatag caagaccgac taccagggct tccaaagcat
     1801 gtacaccttc gtgacttccc atttgccacc cggcttcaac gagtacgact tcgtgcccga
     1861 gagcttcgac cgggacaaaa ccatcgccct gatcatgaac agtagtggca gtaccggatt
     1921 gcccaagggc gtagccctac cgcaccgcac cgcttgtgtc cgattcagtc atgcccgcga
     1981 ccccatcttc ggcaaccaga tcatccccga caccgctatc ctcagcgtgg tgccatttca
     2041 ccacggcttc ggcatgttca ccacgctggg ctacttgatc tgcggctttc gggtcgtgct
     2101 catgtaccgc ttcgaggagg agctattctt gcgcagcttg caagactata agattcaatc
     2161 tgccctgctg gtgcccacac tatttagctt cttcgctaag agcactctca tcgacaagta
     2221 cgacctaagc aacttgcacg agatcgccag cggcggggcg ccgctcagca aggaggtagg
     2281 tgaggccgtg gccaaacgct tccacctacc aggcatccgc cagggctacg gcctgacaga
     2341 aacaaccagc gccattctga tcacccccga aggggacgac aagcctggcg cagtaggcaa
     2401 ggtggtgccc ttcttcgagg ctaaggtggt ggacttggac accggtaaga cactgggtgt
     2461 gaaccagcgc ggcgagctgt gcgtccgtgg ccccatgatc atgagcggct acgttaacaa
     2521 ccccgaggct acaaacgctc tcatcgacaa ggacggctgg ctgcacagcg gcgacatcgc
     2581 ctactgggac gaggacgagc acttcttcat cgtggaccgg ctgaagagcc tgatcaaata
     2641 caagggctac caggtagccc cagccgaact ggagagcatc ctgctgcaac accccaacat
     2701 cttcgacgcc ggggtcgccg gcctgcccga cgacgatgcc ggcgagctgc ccgccgcagt
     2761 cgtcgtgctg gaacacggta aaaccatgac cgagaaggag atcgtggact atgtggccag
     2821 ccaggttaca accgccaaga agctgcgcgg tggtgttgtg ttcgtggacg aggtgcctaa
     2881 aggactgacc ggcaagttgg acgcccgcaa gatccgcgag attctcatta aggccaagaa
     2941 gggcggcaag atcgccgtgt aataattcta gagtcggggc ggccggccgc ttcgagcaga
     3001 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3061 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3121 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     3181 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccgtttgc
     3241 gtattgggcg ctcttccgct gatctgcgca gcaccatggc ctgaaataac ctctgaaaga
     3301 ggaacttggt tagctacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     3361 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3421 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3481 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3541 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3601 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3661 ggcctaggct tttgcaaaaa gctcgattct tctgacacta gcgccaccat gaagaagccc
     3721 gaactcaccg ctaccagcgt tgaaaaattt ctcatcgaga agttcgacag tgtgagcgac
     3781 ctgatgcagt tgtcggaggg cgaagagagc cgagccttca gcttcgatgt cggcggacgc
     3841 ggctatgtac tgcgggtgaa tagctgcgct gatggcttct acaaagaccg ctacgtgtac
     3901 cgccacttcg ccagcgctgc actacccatc cccgaagtgt tggacatcgg cgagttcagc
     3961 gagagcctga catactgcat cagtagacgc gcccaaggcg ttactctcca agacctcccc
     4021 gaaacagagc tgcctgctgt gttacagcct gtcgccgaag ctatggatgc tattgccgcc
     4081 gccgacctca gtcaaaccag cggcttcggc ccattcgggc cccaaggcat cggccagtac
     4141 acaacctggc gggatttcat ttgcgccatt gctgatcccc atgtctacca ctggcagacc
     4201 gtgatggacg acaccgtgtc cgccagcgta gctcaagccc tggacgaact gatgctgtgg
     4261 gccgaagact gtcccgaggt gcgccacctc gtccatgccg acttcggcag caacaacgtc
     4321 ctgaccgaca acggccgcat caccgccgta atcgactggt ccgaagctat gttcggggac
     4381 agtcagtacg aggtggccaa catcttcttc tggcggccct ggctggcttg catggagcag
     4441 cagactcgct acttcgagcg ccggcatccc gagctggccg gcagccctcg tctgcgagcc
     4501 tacatgctgc gcatcggcct ggatcagctc taccagagcc tcgtggacgg caacttcgac
     4561 gatgctgcct gggctcaagg ccgctgcgat gccatcgtcc gcagcggggc cggcaccgtc
     4621 ggtcgcacac aaatcgctcg ccggagcgca gccgtatgga ccgacggctg cgtcgaggtg
     4681 ctggccgaca gcggcaaccg ccggcccagt acacgaccgc gcgctaagga ggtaggtcga
     4741 gtttaaactc tagaaccggt catggccgca ataaaatatc tttattttca ttacatctgt
     4801 gtgttggttt tttgtgtgtt cgaactagat gctgtcgacc gatgcccttg agagccttca
     4861 acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg
     4921 tcttctttat catgcaactc gtaggacagg tgccggcagc gctcttccgc ttcctcgctc
     4981 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5041 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5101 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5161 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5221 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5281 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5341 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5401 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5461 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5521 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5581 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5641 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5701 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5761 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5821 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5881 tatgagtaaa cttggtctga cagcggccgc aaatgctaaa ccactgcagt ggttaccagt
     5941 gcttgatcag tgaggcaccg atctcagcga tctgcctatt tcgttcgtcc atagtggcct
     6001 gactccccgt cgtgtagatc actacgattc gtgagggctt accatcaggc cccagcgcag
     6061 caatgatgcc gcgagagccg cgttcaccgg cccccgattt gtcagcaatg aaccagccag
     6121 cagggagggc cgagcgaaga agtggtcctg ctactttgtc cgcctccatc cagtctatga
     6181 gctgctgtcg tgatgctaga gtaagaagtt cgccagtgag tagtttccga agagttgtgg
     6241 ccattgctac tggcatcgtg gtatcacgct cgtcgttcgg tatggcttcg ttcaactctg
     6301 gttcccagcg gtcaagccgg gtcacatgat cacccatatt atgaagaaat gcagtcagct
     6361 ccttagggcc tccgatcgtt gtcagaagta agttggccgc ggtgttgtcg ctcatggtaa
     6421 tggcagcact acacaattct cttaccgtca tgccatccgt aagatgcttt tccgtgaccg
     6481 gcgagtactc aaccaagtcg ttttgtgagt agtgtatacg gcgaccaagc tgctcttgcc
     6541 cggcgtctat acgggacaac accgcgccac atagcagtac tttgaaagtg ctcatcatcg
     6601 ggaatcgttc ttcggggcgg aaagactcaa ggatcttgcc gctattgaga tccagttcga
     6661 tatagcccac tcttgcaccc agttgatctt cagcatcttt tactttcacc agcgtttcgg
     6721 ggtgtgcaaa aacaggcaag caaaatgccg caaagaaggg aatgagtgcg acacgaaaat
     6781 gttggatgct catactcgtc ctttttcaat attattgaag catttatcag ggttactagt
     6841 acgtctctca aggataagta agtaatatta aggtacggga ggtattggac aggccgcaat
     6901 aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagtacta
     6961 acatacgctc tccatcaaaa caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc
     7021 agtgcaagtg caggtgccag aacatttctc t
//
