LOCUS       sequence-178144-        5145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fascin promoter for luciferase assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5145)
  AUTHORS   Sun J, He H, Pillai S, Xiong Y, Challa S, Xu L, Chellappan S, Yang S
  TITLE     GATA3 transcription factor abrogates Smad4 transcription 
            factor-mediated fascin overexpression, invadopodium formation, and 
            breast cancer cell invasion.
  JOURNAL   J Biol Chem. 2013 Dec 27;288(52):36971-82. doi: 
            10.1074/jbc.M113.506535. Epub 2013 Nov 14.
  PUBMED    24235142
REFERENCE   2  (bases 1 to 5145)
  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: 
            "10.1074/jbc.M113.506535"; journalName: "J Biol Chem"; date: 
            "2013-12-27- 27"; volume: "288"; issue: "52"; pages: "36971-82"
COMMENT     Sequence Label: pGL3-Basic-Fascin-Promoter (-210-0)
FEATURES             Location/Qualifiers
     source          1..5145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..92
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     41..60
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      508..517
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             514..2166
                     /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(580..598)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2207..2328
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2218..2237)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2272..2291
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2576..2593)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2747..3335)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2827..2846)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3506..4297,4298..4366))
                     /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     4129..4148
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4367..4471)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4498..4953
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4585..4604)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4795..4816
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5084..5132
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
       61 cagtgcaagt gcaggtgcca gaacatttct ctatcgatag gtaccgagct cttacgcgtg
      121 ctagcccggg ctcgagggtt gtgaggggtg atgtcctcag gcggcggcgc tgcggggtgc
      181 ggcgaggaca ccggtggggt gagagcaccg gcggggcagc agcgggggcc gcagcgccgg
      241 gtccctcggc ccggggcccc tcccgcgcgg agccaggggc gggacagggg ggcgtggcct
      301 ggtggcgctg acgtcacctc gcctataaaa tgtccggggc gccgctagct gggctttgtg
      361 gagcgctgcg gagggtgcgt gcgggccgcg gcagccgaac aaaggagcag gggcgccgcc
      421 gcagggaccc gccacccacc tcccggggcc gcgcagcggc ctctcgtcta ctgccaccaa
      481 gcttggcatt ccggtactgt tggtaaagcc accatggaag acgccaaaaa cataaagaaa
      541 ggcccggcgc cattctatcc gctggaagat ggaaccgctg gagagcaact gcataaggct
      601 atgaagagat acgccctggt tcctggaaca attgctttta cagatgcaca tatcgaggtg
      661 gacatcactt acgctgagta cttcgaaatg tccgttcggt tggcagaagc tatgaaacga
      721 tatgggctga atacaaatca cagaatcgtc gtatgcagtg aaaactctct tcaattcttt
      781 atgccggtgt tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa cgacatttat
      841 aatgaacgtg aattgctcaa cagtatgggc atttcgcagc ctaccgtggt gttcgtttcc
      901 aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaagc tcccaatcat ccaaaaaatt
      961 attatcatgg attctaaaac ggattaccag ggatttcagt cgatgtacac gttcgtcaca
     1021 tctcatctac ctcccggttt taatgaatac gattttgtgc cagagtcctt cgatagggac
     1081 aagacaattg cactgatcat gaactcctct ggatctactg gtctgcctaa aggtgtcgct
     1141 ctgcctcata gaactgcctg cgtgagattc tcgcatgcca gagatcctat ttttggcaat
     1201 caaatcattc cggatactgc gattttaagt gttgttccat tccatcacgg ttttggaatg
     1261 tttactacac tcggatattt gatatgtgga tttcgagtcg tcttaatgta tagatttgaa
     1321 gaagagctgt ttctgaggag ccttcaggat tacaagattc aaagtgcgct gctggtgcca
     1381 accctattct ccttcttcgc caaaagcact ctgattgaca aatacgattt atctaattta
     1441 cacgaaattg cttctggtgg cgctcccctc tctaaggaag tcggggaagc ggttgccaag
     1501 aggttccatc tgccaggtat caggcaagga tatgggctca ctgagactac atcagctatt
     1561 ctgattacac ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt
     1621 gaagcgaagg ttgtggatct ggataccggg aaaacgctgg gcgttaatca aagaggcgaa
     1681 ctgtgtgtga gaggtcctat gattatgtcc ggttatgtaa acaatccgga agcgaccaac
     1741 gccttgattg acaaggatgg atggctacat tctggagaca tagcttactg ggacgaagac
     1801 gaacacttct tcatcgttga ccgcctgaag tctctgatta agtacaaagg ctatcaggtg
     1861 gctcccgctg aattggaatc catcttgctc caacacccca acatcttcga cgcaggtgtc
     1921 gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt tttggagcac
     1981 ggaaagacga tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt aacaaccgcg
     2041 aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa
     2101 ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca agaagggcgg aaagatcgcc
     2161 gtgtaattct agagtcgggg cggccggccg cttcgagcag acatgataag atacattgat
     2221 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     2281 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     2341 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     2401 aacctctaca aatgtggtaa aatcgataag gatccgtcga ccgatgccct tgagagcctt
     2461 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     2521 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc
     2581 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2641 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2701 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2761 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2821 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2881 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2941 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3001 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3061 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3121 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3181 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3241 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3301 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3361 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3421 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3481 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3541 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3601 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3661 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3721 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3781 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3841 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3901 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3961 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4021 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4081 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4141 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4201 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4261 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4321 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4381 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4441 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4501 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4561 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4621 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4681 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4741 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4801 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4861 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4921 cgaattttaa caaaatatta acgcttacaa tttgccattc gccattcagg ctgcgcaact
     4981 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagcccaag ctaccatgat
     5041 aagtaagtaa tattaaggta cgggaggtac ttggagcggc cgcaataaaa tatctttatt
     5101 ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gtact
//
