LOCUS       sequence-122537-        5005 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  basal plasmid containing minimal fgf4 promoter used for testing 
            enhancer activity of DNAs inserted immediately upstream of the 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5005)
  AUTHORS   Murtha M, Tokcaer-Keskin Z, Tang Z, Strino F, Chen X, Wang Y, Xi X, 
            Basilico C, Brown S, Bonneau R, Kluger Y, Dailey L
  TITLE     FIREWACh: high-throughput functional detection of transcriptional 
            regulatory modules in mammalian cells.
  JOURNAL   Nat Methods. 2014 May;11(5):559-65. doi: 10.1038/nmeth.2885. Epub 
            2014 Mar 23.
  PUBMED    24658142
REFERENCE   2  (bases 1 to 5005)
  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: 
            "10.1038/nmeth.2885"; journalName: "Nat Methods"; date: "2014-05"; 
            volume: "11"; issue: "5"; pages: "559-65"
COMMENT     Sequence Label: fgfprom luc
FEATURES             Location/Qualifiers
     source          1..5005
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      269..278
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             275..1927
                     /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(341..359)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1968..2089
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1979..1998)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2033..2052
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2337..2354)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2508..3096)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2588..2607)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3267..4058,4059..4127))
                     /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     3890..3909
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4128..4232)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4259..4714
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4346..4365)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4556..4577
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4845..4893
                     /note="synthetic polyadenylation signal"
     misc_feature    4907..4998
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4947..4966
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcggagat ctcgagctct tacgcgtgct
       61 agcccagcac ccgggcgcgg ggcaggcgag taggaggggg cgccggctat atataccact
      121 gctccggagg gctgggcgcg cggggactat cccgccaccg ttgcgtccct atttgctctc
      181 gctacttagg tctgtgcgca gcactaccga actcacggcc cgcagctcga actcacgcaa
      241 agcttggcat tccggtactg ttggtaaagc caccatggaa gacgccaaaa acataaagaa
      301 aggcccggcg ccattctatc cgctggaaga tggaaccgct ggagagcaac tgcataaggc
      361 tatgaagaga tacgccctgg ttcctggaac aattgctttt acagatgcac atatcgaggt
      421 ggacatcact tacgctgagt acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg
      481 atatgggctg aatacaaatc acagaatcgt cgtatgcagt gaaaactctc ttcaattctt
      541 tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga acgacattta
      601 taatgaacgt gaattgctca acagtatggg catttcgcag cctaccgtgg tgttcgtttc
      661 caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat
      721 tattatcatg gattctaaaa cggattacca gggatttcag tcgatgtaca cgttcgtcac
      781 atctcatcta cctcccggtt ttaatgaata cgattttgtg ccagagtcct tcgataggga
      841 caagacaatt gcactgatca tgaactcctc tggatctact ggtctgccta aaggtgtcgc
      901 tctgcctcat agaactgcct gcgtgagatt ctcgcatgcc agagatccta tttttggcaa
      961 tcaaatcatt ccggatactg cgattttaag tgttgttcca ttccatcacg gttttggaat
     1021 gtttactaca ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt atagatttga
     1081 agaagagctg tttctgagga gccttcagga ttacaagatt caaagtgcgc tgctggtgcc
     1141 aaccctattc tccttcttcg ccaaaagcac tctgattgac aaatacgatt tatctaattt
     1201 acacgaaatt gcttctggtg gcgctcccct ctctaaggaa gtcggggaag cggttgccaa
     1261 gaggttccat ctgccaggta tcaggcaagg atatgggctc actgagacta catcagctat
     1321 tctgattaca cccgaggggg atgataaacc gggcgcggtc ggtaaagttg ttccattttt
     1381 tgaagcgaag gttgtggatc tggataccgg gaaaacgctg ggcgttaatc aaagaggcga
     1441 actgtgtgtg agaggtccta tgattatgtc cggttatgta aacaatccgg aagcgaccaa
     1501 cgccttgatt gacaaggatg gatggctaca ttctggagac atagcttact gggacgaaga
     1561 cgaacacttc ttcatcgttg accgcctgaa gtctctgatt aagtacaaag gctatcaggt
     1621 ggctcccgct gaattggaat ccatcttgct ccaacacccc aacatcttcg acgcaggtgt
     1681 cgcaggtctt cccgacgatg acgccggtga acttcccgcc gccgttgttg ttttggagca
     1741 cggaaagacg atgacggaaa aagagatcgt ggattacgtc gccagtcaag taacaaccgc
     1801 gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa
     1861 actcgacgca agaaaaatca gagagatcct cataaaggcc aagaagggcg gaaagatcgc
     1921 cgtgtaattc tagagtcggg gcggccggcc gcttcgagca gacatgataa gatacattga
     1981 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     2041 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     2101 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     2161 aaacctctac aaatgtggta aaatcgataa ggatccgtcg accgatgccc ttgagagcct
     2221 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
     2281 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg
     2341 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2401 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2461 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2521 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2581 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2641 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2701 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2761 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2821 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2881 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2941 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3001 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3061 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3121 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3181 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3241 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3301 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3361 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3421 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3481 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3541 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3601 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3661 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3721 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3781 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3841 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3901 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3961 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4021 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4081 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4141 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4201 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     4261 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4321 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4381 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4441 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4501 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4561 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4621 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4681 gcgaatttta acaaaatatt aacgtttaca atttcccatt cgccattcag gctgcgcaac
     4741 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagcccaa gctaccatga
     4801 taagtaagta atattaaggt acgggaggta cttggagcgg ccgcaataaa atatctttat
     4861 tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca tacgctctcc
     4921 atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt gcaagtgcag
     4981 gtgccagaac atttctctat cgata
//
