LOCUS       sequence-109372-        6561 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase expression driven by Lin28b promoter P3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6561)
  AUTHORS   Chang TC, Zeitels LR, Hwang HW, Chivukula RR, Wentzel EA, Dews M, 
            Jung J, Gao P, Dang CV, Beer MA, Thomas-Tikhonenko A, Mendell JT
  TITLE     Lin-28B transactivation is necessary for Myc-mediated let-7 
            repression and proliferation.
  JOURNAL   Proc Natl Acad Sci U S A. 2009 Mar 3;106(9):3384-9. doi: 
            10.1073/pnas.0808300106. Epub 2009 Feb 11.
  PUBMED    19211792
REFERENCE   2  (bases 1 to 6561)
  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: 
            "10.1073/pnas.0808300106"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2009-03-3- 3"; volume: "106"; issue: "9"; pages: "3384-9"
COMMENT     Sequence Label: pGL3-IRES-Lin28b-P3
FEATURES             Location/Qualifiers
     source          1..6561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1212..1779
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1373..1390)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1600..1619
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      1825..1834
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1831..3483
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(1897..1915)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3524..3645
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3535..3554)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3589..3608
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3893..3910)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4064..4652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4144..4163)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4823..5614,5615..5683))
                     /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     5446..5465
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5684..5788)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5815..6270
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5902..5921)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6112..6133
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6401..6449
                     /note="synthetic polyadenylation signal"
     misc_feature    6463..6554
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6503..6522
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctggagtgt ggatgtgttt gaagagactg ggagggggat
       61 tgctggcatg gtgggggagg gtcaactcgg tagctttgtc gaaggaagtg gtatctgaac
      121 agttagtttc ttggtaaact gcttgtttaa gaaaatgggg ggtggggtag gcaagtctgg
      181 tggtgcattt taaggtttta aaatagcgat attttatatt ttaaaggaaa taaagtcttt
      241 ttagttaatc tataatattt attatagtat tcgataaggg cagaaatact gttgccagca
      301 tcttgtaatc aatcgtttga accagtttga aaggtaaagg gggtaggcat tttgaaactg
      361 ggggtcagtt gaggaaggac agtaggagct tcatccgttg ttacattttc gtttattcaa
      421 ctcttctagc cattggtagt ggacttttaa aatgtcaggg tacaagtcaa tatgatatga
      481 atttaccttt tttctcccac ttgtccttct ttctccaccc caccccctgc ccgcgccggc
      541 tttttttctc tcccacctct ttctttgact ctgctgccgc ctatcctggc ggctcctctt
      601 cacctcccct gcctggcgat cccctccccc tcctgcccta tgtcctccag ctcttgggga
      661 acaatcgcca ctgattgagg ttcactctat gttaggctgg aaaactgggc tgttatttag
      721 gaagtcatta atcacatctc ctcccccgga aagagatggc agagaaacct gtgaagtaca
      781 attccttacc aaccccctct aaaattttcc aaaatggttt acctaaacac aatttgacta
      841 tatgaaggtg aaaacgattt ttctagtagg attttatttt aaatttgaaa aaatttcggg
      901 cataatcagg taatttatgt gcctctgcct tttcctaggt gtatgttttc attatctaaa
      961 gtttcatggg gatgtaatta ggtttaatta gatcttttat aaattatata ttttcctcta
     1021 agaattgaat taaatatgaa attagcgttc caagcaggtt tactgccatg gaatagctga
     1081 attcttaagg ggaaaaaaag cgttaacatc ggttgtttta tttaaacgac cccctcccat
     1141 gattttagtc aggcttgtgt gtaaggcacg cgtgctagcc cgggctcgag atctgacccc
     1201 atgcatcgaa ttccgccccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     1261 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     1321 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     1381 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     1441 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     1501 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     1561 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     1621 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     1681 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     1741 cacggggacg tggttttcct ttgaaaaaca cgatgataag atctgcgatc taagtaagct
     1801 tggcattccg gtactgttgg taaagccacc atggaagacg ccaaaaacat aaagaaaggc
     1861 ccggcgccat tctatccgct ggaagatgga accgctggag agcaactgca taaggctatg
     1921 aagagatacg ccctggttcc tggaacaatt gcttttacag atgcacatat cgaggtggac
     1981 atcacttacg ctgagtactt cgaaatgtcc gttcggttgg cagaagctat gaaacgatat
     2041 gggctgaata caaatcacag aatcgtcgta tgcagtgaaa actctcttca attctttatg
     2101 ccggtgttgg gcgcgttatt tatcggagtt gcagttgcgc ccgcgaacga catttataat
     2161 gaacgtgaat tgctcaacag tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa
     2221 aaggggttgc aaaaaatttt gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt
     2281 atcatggatt ctaaaacgga ttaccaggga tttcagtcga tgtacacgtt cgtcacatct
     2341 catctacctc ccggttttaa tgaatacgat tttgtgccag agtccttcga tagggacaag
     2401 acaattgcac tgatcatgaa ctcctctgga tctactggtc tgcctaaagg tgtcgctctg
     2461 cctcatagaa ctgcctgcgt gagattctcg catgccagag atcctatttt tggcaatcaa
     2521 atcattccgg atactgcgat tttaagtgtt gttccattcc atcacggttt tggaatgttt
     2581 actacactcg gatatttgat atgtggattt cgagtcgtct taatgtatag atttgaagaa
     2641 gagctgtttc tgaggagcct tcaggattac aagattcaaa gtgcgctgct ggtgccaacc
     2701 ctattctcct tcttcgccaa aagcactctg attgacaaat acgatttatc taatttacac
     2761 gaaattgctt ctggtggcgc tcccctctct aaggaagtcg gggaagcggt tgccaagagg
     2821 ttccatctgc caggtatcag gcaaggatat gggctcactg agactacatc agctattctg
     2881 attacacccg agggggatga taaaccgggc gcggtcggta aagttgttcc attttttgaa
     2941 gcgaaggttg tggatctgga taccgggaaa acgctgggcg ttaatcaaag aggcgaactg
     3001 tgtgtgagag gtcctatgat tatgtccggt tatgtaaaca atccggaagc gaccaacgcc
     3061 ttgattgaca aggatggatg gctacattct ggagacatag cttactggga cgaagacgaa
     3121 cacttcttca tcgttgaccg cctgaagtct ctgattaagt acaaaggcta tcaggtggct
     3181 cccgctgaat tggaatccat cttgctccaa caccccaaca tcttcgacgc aggtgtcgca
     3241 ggtcttcccg acgatgacgc cggtgaactt cccgccgccg ttgttgtttt ggagcacgga
     3301 aagacgatga cggaaaaaga gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa
     3361 aagttgcgcg gaggagttgt gtttgtggac gaagtaccga aaggtcttac cggaaaactc
     3421 gacgcaagaa aaatcagaga gatcctcata aaggccaaga agggcggaaa gatcgccgtg
     3481 taattctaga gtcggggcgg ccggccgctt cgagcagaca tgataagata cattgatgag
     3541 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     3601 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     3661 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     3721 ctctacaaat gtggtaaaat cgataaggat ccgtcgaccg atgcccttga gagccttcaa
     3781 cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt
     3841 cttctttatc atgcaactcg taggacaggt gccggcagcg ctcttccgct tcctcgctca
     3901 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3961 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4021 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4081 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4141 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4201 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4261 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4321 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4381 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4441 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4501 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4561 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4621 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4681 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4741 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4801 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4861 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4921 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4981 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5041 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5101 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5161 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5221 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5281 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5341 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5401 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5461 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5521 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5581 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5641 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5701 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5761 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc
     5821 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5881 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5941 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     6001 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     6061 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     6121 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     6181 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     6241 attttaacaa aatattaacg cttacaattt gccattcgcc attcaggctg cgcaactgtt
     6301 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gcccaagcta ccatgataag
     6361 taagtaatat taaggtacgg gaggtacttg gagcggccgc aataaaatat ctttattttc
     6421 attacatctg tgtgttggtt ttttgtgtga atcgatagta ctaacatacg ctctccatca
     6481 aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc
     6541 cagaacattt ctctatcgat a
//
