LOCUS       sequence-232893-        5317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For luciferase expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5317)
  AUTHORS   Miao YL, Gambini A, Zhang Y, Padilla-Banks E, Jefferson WN, 
            Bernhardt ML, Huang W, Li L, Williams CJ
  TITLE     Mediator complex component MED13 regulates zygotic genome activation
            and is required for postimplantation development in the mouse.
  JOURNAL   Biol Reprod. 2018 Jan 9. pii: 4794936. doi: 10.1093/biolre/ioy004.
  PUBMED    29325037
REFERENCE   2  (bases 1 to 5317)
  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: "Biol 
            Reprod. 2018 Jan 9. pii: 4794936. doi: 10.1093/biolre/ioy004."
COMMENT     Sequence Label: pF208wt-luc
FEATURES             Location/Qualifiers
     source          1..5317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1169..1178
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1175..2827
                     /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    2871..2992
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2882..2901)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2936..2955
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3240..3257)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3411..3999)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3491..3510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4199..4990,4991..5059))
                     /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    5164..5212
                     /note="synthetic polyadenylation signal"
     misc_feature    5226..5317
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5266..5285
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcctcgagta aatactttgg atgaaattag
       61 acctggcttt tgtcatctca atttgcctaa atttcaacat ttaaattgtg taaaagcatc
      121 tacttggtat ctgttgtgag aatgtgtcta tacagatggt aactcagaac cccttaccta
      181 gaaagaaact attgaaaggc cctcccttca tttcactatc aaattgtttt cttctaggtg
      241 gtcgaacatt cttctaggtg tgcgagcatt cgtgcctccc atcattaggt tgtaaattgt
      301 gcggaagaat cccaaatatt gaatgtttgc ttatgtagtg ctttggtctg gagtacgacc
      361 agtgctcgtg atctactggg aaagcagaaa gtctggatgg aataggatgt tatcaccctg
      421 cttatcactg gaggccattt tcttatctct cttcactgcc tttcttattg tttgtttata
      481 atctttatgc atataggtgg cttgcctgca tttatgatag agcaccacac gtatacctag
      541 tgccctccga ggccagaaaa ggacttggat tctttgaagt gactaactgc tggttgagtt
      601 gtcctatgtc ggctggagct cgaattctgg tcctttggaa gatcagctca gagcagaccg
      661 atcacacgac caggatcagc caattatgtg ccgggtttgg tccacagacc tccggaggaa
      721 agaaggtgct tacttgcaag agtactgtcc gaaagacgtc ctctgggctt gcctccgtca
      781 aacgcagtgc ggagagggca gggcccagga atccgcagaa tgctgccaag cccaagccct
      841 cactctgaag cgatggcgtc gacgctttcc ccgggcgtgg cactctcctc ttttggcggg
      901 ctcggcatcg cgaggctccg ccctcctgga acggtctctc tgcagtcgcg gaggcgggct
      961 ctgcatttct attggtagag cgggtccgcc gaggccccgc cccctcccgg gcccgacgcg
     1021 agcacgttgg gtagccaggg ccggttgaaa agccgcccga atccggagca gctcccgggg
     1081 cgagaggcac gcttagggcc gcggtggcgg ttaagaggcg tctctgtggc tggagtgcga
     1141 agcttggcaa tccggtactg ttggtaaagc caccatggaa gatgccaaaa acattaagaa
     1201 gggcccagcg ccattctacc cactcgaaga cgggaccgcc ggcgagcagc tgcacaaagc
     1261 catgaagcgc tacgccctgg tgcccggcac catcgccttt accgacgcac atatcgaggt
     1321 ggacattacc tacgccgagt acttcgagat gagcgttcgg ctggcagaag ctatgaagcg
     1381 ctatgggctg aatacaaacc atcggatcgt ggtgtgcagc gagaatagct tgcagttctt
     1441 catgcccgtg ttgggtgccc tgttcatcgg tgtggctgtg gccccagcta acgacatcta
     1501 caacgagcgc gagctgctga acagcatggg catcagccag cccaccgtcg tattcgtgag
     1561 caagaaaggg ctgcaaaaga tcctcaacgt gcaaaagaag ctaccgatca tacaaaagat
     1621 catcatcatg gatagcaaga ccgactacca gggcttccaa agcatgtaca ccttcgtgac
     1681 ttcccatttg ccacccggct tcaacgagta cgacttcgtg cccgagagct tcgaccggga
     1741 caaaaccatc gccctgatca tgaacagtag tggcagtacc ggattgccca agggcgtagc
     1801 cctaccgcac cgcaccgctt gtgtccgatt cagtcatgcc cgcgacccca tcttcggcaa
     1861 ccagatcatc cccgacaccg ctatcctcag cgtggtgcca tttcaccacg gcttcggcat
     1921 gttcaccacg ctgggctact tgatctgcgg ctttcgggtc gtgctcatgt accgcttcga
     1981 ggaggagcta ttcttgcgca gcttgcaaga ctataagatt caatctgccc tgctggtgcc
     2041 cacactattt agcttcttcg ctaagagcac tctcatcgac aagtacgacc taagcaactt
     2101 gcacgagatc gccagcggcg gggcgccgct cagcaaggag gtaggtgagg ccgtggccaa
     2161 acgcttccac ctaccaggca tccgccaggg ctacggcctg acagaaacaa ccagcgccat
     2221 tctgatcacc cccgaagggg acgacaagcc tggcgcagta ggcaaggtgg tgcccttctt
     2281 cgaggctaag gtggtggact tggacaccgg taagacactg ggtgtgaacc agcgcggcga
     2341 gctgtgcgtc cgtggcccca tgatcatgag cggctacgtt aacaaccccg aggctacaaa
     2401 cgctctcatc gacaaggacg gctggctgca cagcggcgac atcgcctact gggacgagga
     2461 cgagcacttc ttcatcgtgg accggctgaa gagcctgatc aaatacaagg gctaccaggt
     2521 agccccagcc gaactggaga gcatcctgct gcaacacccc aacatcttcg acgccggggt
     2581 cgccggcctg cccgacgacg atgccggcga gctgcccgcc gcagtcgtcg tgctggaaca
     2641 cggtaaaacc atgaccgaga aggagatcgt ggactatgtg gccagccagg ttacaaccgc
     2701 caagaagctg cgcggtggtg ttgtgttcgt ggacgaggtg cctaaaggac tgaccggcaa
     2761 gttggacgcc cgcaagatcc gcgagattct cattaaggcc aagaagggcg gcaagatcgc
     2821 cgtgtaataa ttctagagtc ggggcggccg gccgcttcga gcagacatga taagatacat
     2881 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2941 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     3001 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     3061 gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg tcgaccgatg cccttgagag
     3121 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta
     3181 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc
     3241 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3301 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3361 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3421 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3481 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3541 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3601 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3661 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3721 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3781 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3841 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3901 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3961 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4021 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4081 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4141 agtatatatg agtaaacttg gtctgacagc ggccgcaaat gctaaaccac tgcagtggtt
     4201 accagtgctt gatcagtgag gcaccgatct cagcgatctg cctatttcgt tcgtccatag
     4261 tggcctgact ccccgtcgtg tagatcacta cgattcgtga gggcttacca tcaggcccca
     4321 gcgcagcaat gatgccgcga gagccgcgtt caccggcccc cgatttgtca gcaatgaacc
     4381 agccagcagg gagggccgag cgaagaagtg gtcctgctac tttgtccgcc tccatccagt
     4441 ctatgagctg ctgtcgtgat gctagagtaa gaagttcgcc agtgagtagt ttccgaagag
     4501 ttgtggccat tgctactggc atcgtggtat cacgctcgtc gttcggtatg gcttcgttca
     4561 actctggttc ccagcggtca agccgggtca catgatcacc catattatga agaaatgcag
     4621 tcagctcctt agggcctccg atcgttgtca gaagtaagtt ggccgcggtg ttgtcgctca
     4681 tggtaatggc agcactacac aattctctta ccgtcatgcc atccgtaaga tgcttttccg
     4741 tgaccggcga gtactcaacc aagtcgtttt gtgagtagtg tatacggcga ccaagctgct
     4801 cttgcccggc gtctatacgg gacaacaccg cgccacatag cagtactttg aaagtgctca
     4861 tcatcgggaa tcgttcttcg gggcggaaag actcaaggat cttgccgcta ttgagatcca
     4921 gttcgatata gcccactctt gcacccagtt gatcttcagc atcttttact ttcaccagcg
     4981 tttcggggtg tgcaaaaaca ggcaagcaaa atgccgcaaa gaagggaatg agtgcgacac
     5041 gaaaatgttg gatgctcata ctcgtccttt ttcaatatta ttgaagcatt tatcagggtt
     5101 actagtacgt ctctcaagga taagtaagta atattaaggt acgggaggta ttggacaggc
     5161 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata
     5221 gtactaacat acgctctcca tcaaaacaaa acgaaacaaa acaaactagc aaaataggct
     5281 gtccccagtg caagtgcagg tgccagaaca tttctct
//
