LOCUS       sequence-451020-        6760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  0.9 kb from mouse Igf2 promoter in front of firefly luciferase 
            reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6760)
  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 6760)
  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-Igf2Promoter
FEATURES             Location/Qualifiers
     source          1..6760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        955..986
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      1013..1022
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1019..2671
                     /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    2715..2836
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2726..2745)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2780..2799
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3030..3387
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3030..3221
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3053..3073)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3238..3373
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3300..3319
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3418..4455
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4434..4442
                     /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    4479..4527
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4683..4700)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4854..5442)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4934..4953)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5642..6433,6434..6502))
                     /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    6607..6655
                     /note="synthetic polyadenylation signal"
     misc_feature    6669..6760
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6709..6728
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcctcgagtt gcgggggcgg tcccggggcg
       61 gggcgagggc cctgcggacg cccattggcg cgggcgtaag gccagcgggg cccgagcggg
      121 cgccgagccg cggggtggcg cggctataag aaccgggcgt tggcgcccgg agttcgcctg
      181 ctctccggcg gagctgcgtg aggccaggcc ggcccccggc ccccccttcc ggccgccccc
      241 gcctcctggc ccacgcccgc ccgcgctcgg cccgccagcg cctccatccg ggctggcggc
      301 cccgcgtcga cgccgtccgc cacctcgctg ctgctaactc ctgtgcaggg cgccgtcggc
      361 ggggccgcgc tccgtcgggc ctgcggatct ccccaccgcc tcctcctcta tctacctcaa
      421 caccccattc ctgcttcgcc agaggaggcg gtccccaccg caggcagtcc ggcttgcagg
      481 tcgccggcgt tgtcatcccc cgcgcttccc ctcccagccc tccccggtgc gcagcccggc
      541 tgctcccctc ttttcgctgc agtcccgagc agctgaggcg ccgccacgcc tgtccccccc
      601 acaagaagcc cgggcttacg acggctgagg gctccgtcga ccctaaccga gctgggtgcc
      661 cgtggccggg gtgacgcctc cattcctccc ccctcaacac cgtcctcggt ccttcgaagt
      721 tgcatcctct cctctgctta gggtgcgccc cccctcgcgc acccgcttac ccgccacctt
      781 tcctaagctc ccctcctgcc ccctccccgt tcctcctcgc ctcagactcc ctccccctca
      841 cgtccgccct ctgccttcgc ctacccaagt ggattaatta tacgctttct gtttctctcc
      901 gtgctgtcct ctcccgctgt gagcctaccc gcctctcgct gtaagcttag acactagagg
      961 gtatataatg gaagctcgac ttccagcttg gcaatccggt actgttggta aagccaccat
     1021 ggaagatgcc aaaaacatta agaagggccc agcgccattc tacccactcg aagacgggac
     1081 cgccggcgag cagctgcaca aagccatgaa gcgctacgcc ctggtgcccg gcaccatcgc
     1141 ctttaccgac gcacatatcg aggtggacat tacctacgcc gagtacttcg agatgagcgt
     1201 tcggctggca gaagctatga agcgctatgg gctgaataca aaccatcgga tcgtggtgtg
     1261 cagcgagaat agcttgcagt tcttcatgcc cgtgttgggt gccctgttca tcggtgtggc
     1321 tgtggcccca gctaacgaca tctacaacga gcgcgagctg ctgaacagca tgggcatcag
     1381 ccagcccacc gtcgtattcg tgagcaagaa agggctgcaa aagatcctca acgtgcaaaa
     1441 gaagctaccg atcatacaaa agatcatcat catggatagc aagaccgact accagggctt
     1501 ccaaagcatg tacaccttcg tgacttccca tttgccaccc ggcttcaacg agtacgactt
     1561 cgtgcccgag agcttcgacc gggacaaaac catcgccctg atcatgaaca gtagtggcag
     1621 taccggattg cccaagggcg tagccctacc gcaccgcacc gcttgtgtcc gattcagtca
     1681 tgcccgcgac cccatcttcg gcaaccagat catccccgac accgctatcc tcagcgtggt
     1741 gccatttcac cacggcttcg gcatgttcac cacgctgggc tacttgatct gcggctttcg
     1801 ggtcgtgctc atgtaccgct tcgaggagga gctattcttg cgcagcttgc aagactataa
     1861 gattcaatct gccctgctgg tgcccacact atttagcttc ttcgctaaga gcactctcat
     1921 cgacaagtac gacctaagca acttgcacga gatcgccagc ggcggggcgc cgctcagcaa
     1981 ggaggtaggt gaggccgtgg ccaaacgctt ccacctacca ggcatccgcc agggctacgg
     2041 cctgacagaa acaaccagcg ccattctgat cacccccgaa ggggacgaca agcctggcgc
     2101 agtaggcaag gtggtgccct tcttcgaggc taaggtggtg gacttggaca ccggtaagac
     2161 actgggtgtg aaccagcgcg gcgagctgtg cgtccgtggc cccatgatca tgagcggcta
     2221 cgttaacaac cccgaggcta caaacgctct catcgacaag gacggctggc tgcacagcgg
     2281 cgacatcgcc tactgggacg aggacgagca cttcttcatc gtggaccggc tgaagagcct
     2341 gatcaaatac aagggctacc aggtagcccc agccgaactg gagagcatcc tgctgcaaca
     2401 ccccaacatc ttcgacgccg gggtcgccgg cctgcccgac gacgatgccg gcgagctgcc
     2461 cgccgcagtc gtcgtgctgg aacacggtaa aaccatgacc gagaaggaga tcgtggacta
     2521 tgtggccagc caggttacaa ccgccaagaa gctgcgcggt ggtgttgtgt tcgtggacga
     2581 ggtgcctaaa ggactgaccg gcaagttgga cgcccgcaag atccgcgaga ttctcattaa
     2641 ggccaagaag ggcggcaaga tcgccgtgta ataattctag agtcggggcg gccggccgct
     2701 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     2761 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     2821 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     2881 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga
     2941 tccgtttgcg tattgggcgc tcttccgctg atctgcgcag caccatggcc tgaaataacc
     3001 tctgaaagag gaacttggtt agctaccttc tgaggcggaa agaaccagct gtggaatgtg
     3061 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3121 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3181 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3241 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3301 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     3361 ttttttggag gcctaggctt ttgcaaaaag ctcgattctt ctgacactag cgccaccatg
     3421 aagaagcccg aactcaccgc taccagcgtt gaaaaatttc tcatcgagaa gttcgacagt
     3481 gtgagcgacc tgatgcagtt gtcggagggc gaagagagcc gagccttcag cttcgatgtc
     3541 ggcggacgcg gctatgtact gcgggtgaat agctgcgctg atggcttcta caaagaccgc
     3601 tacgtgtacc gccacttcgc cagcgctgca ctacccatcc ccgaagtgtt ggacatcggc
     3661 gagttcagcg agagcctgac atactgcatc agtagacgcg cccaaggcgt tactctccaa
     3721 gacctccccg aaacagagct gcctgctgtg ttacagcctg tcgccgaagc tatggatgct
     3781 attgccgccg ccgacctcag tcaaaccagc ggcttcggcc cattcgggcc ccaaggcatc
     3841 ggccagtaca caacctggcg ggatttcatt tgcgccattg ctgatcccca tgtctaccac
     3901 tggcagaccg tgatggacga caccgtgtcc gccagcgtag ctcaagccct ggacgaactg
     3961 atgctgtggg ccgaagactg tcccgaggtg cgccacctcg tccatgccga cttcggcagc
     4021 aacaacgtcc tgaccgacaa cggccgcatc accgccgtaa tcgactggtc cgaagctatg
     4081 ttcggggaca gtcagtacga ggtggccaac atcttcttct ggcggccctg gctggcttgc
     4141 atggagcagc agactcgcta cttcgagcgc cggcatcccg agctggccgg cagccctcgt
     4201 ctgcgagcct acatgctgcg catcggcctg gatcagctct accagagcct cgtggacggc
     4261 aacttcgacg atgctgcctg ggctcaaggc cgctgcgatg ccatcgtccg cagcggggcc
     4321 ggcaccgtcg gtcgcacaca aatcgctcgc cggagcgcag ccgtatggac cgacggctgc
     4381 gtcgaggtgc tggccgacag cggcaaccgc cggcccagta cacgaccgcg cgctaaggag
     4441 gtaggtcgag tttaaactct agaaccggtc atggccgcaa taaaatatct ttattttcat
     4501 tacatctgtg tgttggtttt ttgtgtgttc gaactagatg ctgtcgaccg atgcccttga
     4561 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
     4621 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctcttccgct
     4681 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4741 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4801 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4861 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4921 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4981 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5041 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5101 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5161 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5221 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5281 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5341 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5401 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5461 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5521 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5581 taaagtatat atgagtaaac ttggtctgac agcggccgca aatgctaaac cactgcagtg
     5641 gttaccagtg cttgatcagt gaggcaccga tctcagcgat ctgcctattt cgttcgtcca
     5701 tagtggcctg actccccgtc gtgtagatca ctacgattcg tgagggctta ccatcaggcc
     5761 ccagcgcagc aatgatgccg cgagagccgc gttcaccggc ccccgatttg tcagcaatga
     5821 accagccagc agggagggcc gagcgaagaa gtggtcctgc tactttgtcc gcctccatcc
     5881 agtctatgag ctgctgtcgt gatgctagag taagaagttc gccagtgagt agtttccgaa
     5941 gagttgtggc cattgctact ggcatcgtgg tatcacgctc gtcgttcggt atggcttcgt
     6001 tcaactctgg ttcccagcgg tcaagccggg tcacatgatc acccatatta tgaagaaatg
     6061 cagtcagctc cttagggcct ccgatcgttg tcagaagtaa gttggccgcg gtgttgtcgc
     6121 tcatggtaat ggcagcacta cacaattctc ttaccgtcat gccatccgta agatgctttt
     6181 ccgtgaccgg cgagtactca accaagtcgt tttgtgagta gtgtatacgg cgaccaagct
     6241 gctcttgccc ggcgtctata cgggacaaca ccgcgccaca tagcagtact ttgaaagtgc
     6301 tcatcatcgg gaatcgttct tcggggcgga aagactcaag gatcttgccg ctattgagat
     6361 ccagttcgat atagcccact cttgcaccca gttgatcttc agcatctttt actttcacca
     6421 gcgtttcggg gtgtgcaaaa acaggcaagc aaaatgccgc aaagaaggga atgagtgcga
     6481 cacgaaaatg ttggatgctc atactcgtcc tttttcaata ttattgaagc atttatcagg
     6541 gttactagta cgtctctcaa ggataagtaa gtaatattaa ggtacgggag gtattggaca
     6601 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
     6661 atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag
     6721 gctgtcccca gtgcaagtgc aggtgccaga acatttctct
//
