LOCUS       sequence-178143-        5337 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 5337)
  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 5337)
  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 402 mutation
FEATURES             Location/Qualifiers
     source          1..5337
                     /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      700..709
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             706..2358
                     /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(772..790)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2399..2520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2410..2429)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2464..2483
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2768..2785)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2939..3527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3019..3038)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3698..4489,4490..4558))
                     /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     4321..4340
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4559..4663)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4690..5145
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4777..4796)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4987..5008
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5276..5324
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
       61 cagtgcaagt gcaggtgcca gaacatttct ctatcgatag gtaccgagct cttacgcgtg
      121 ctagcccggg ctcgaggtgc tccaaggcca cctcctccag gcagcccttt agtccccacc
      181 ttcgcccagt accgatctgc accgtggtct ctgcagtctc catcgtgacc tcaaacctgg
      241 ctcgtccttg ctcctagcga ggcttggggt cggggtgtcc gaggtggggg acatccgggg
      301 gggttaggtg gctggcgcgg ggagccgggg ttgtgagggg tgatgtcctc aggcggcggc
      361 gctgcggggt gcggcgagga caccggtggg gtgagagcac cggcggggca gcagcggggg
      421 ccgcagcgcc gggtccctcg gcccggggcc cctcccgcgc ggagccaggg gcgggacagg
      481 ggggcgtggc ctggtggcgc tgacgtcacc tcgcctataa aatgtccggg gcgccgctag
      541 ctgggctttg tggagcgctg cggagggtgc gtgcgggccg cggcagccga acaaaggagc
      601 aggggcgccg ccgcagggac ccgccaccca cctcccgggg ccgcgcagcg gcctctcgtc
      661 tactgccacc aagcttggca ttccggtact gttggtaaag ccaccatgga agacgccaaa
      721 aacataaaga aaggcccggc gccattctat ccgctggaag atggaaccgc tggagagcaa
      781 ctgcataagg ctatgaagag atacgccctg gttcctggaa caattgcttt tacagatgca
      841 catatcgagg tggacatcac ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa
      901 gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag tgaaaactct
      961 cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg
     1021 aacgacattt ataatgaacg tgaattgctc aacagtatgg gcatttcgca gcctaccgtg
     1081 gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc
     1141 atccaaaaaa ttattatcat ggattctaaa acggattacc agggatttca gtcgatgtac
     1201 acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt gccagagtcc
     1261 ttcgataggg acaagacaat tgcactgatc atgaactcct ctggatctac tggtctgcct
     1321 aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc cagagatcct
     1381 atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc attccatcac
     1441 ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt cgtcttaatg
     1501 tatagatttg aagaagagct gtttctgagg agccttcagg attacaagat tcaaagtgcg
     1561 ctgctggtgc caaccctatt ctccttcttc gccaaaagca ctctgattga caaatacgat
     1621 ttatctaatt tacacgaaat tgcttctggt ggcgctcccc tctctaagga agtcggggaa
     1681 gcggttgcca agaggttcca tctgccaggt atcaggcaag gatatgggct cactgagact
     1741 acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt cggtaaagtt
     1801 gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct gggcgttaat
     1861 caaagaggcg aactgtgtgt gagaggtcct atgattatgt ccggttatgt aaacaatccg
     1921 gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga catagcttac
     1981 tgggacgaag acgaacactt cttcatcgtt gaccgcctga agtctctgat taagtacaaa
     2041 ggctatcagg tggctcccgc tgaattggaa tccatcttgc tccaacaccc caacatcttc
     2101 gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg aacttcccgc cgccgttgtt
     2161 gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt cgccagtcaa
     2221 gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt accgaaaggt
     2281 cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc caagaagggc
     2341 ggaaagatcg ccgtgtaatt ctagagtcgg ggcggccggc cgcttcgagc agacatgata
     2401 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2461 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2521 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     2581 taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccgtc gaccgatgcc
     2641 cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc
     2701 cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt
     2761 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2821 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2881 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2941 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3001 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3061 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3121 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3181 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3241 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3301 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3361 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3421 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3481 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3541 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3601 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3661 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3721 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3781 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3841 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3901 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3961 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4021 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4081 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4141 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4201 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4261 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4321 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4381 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4441 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4501 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4561 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4621 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4681 tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4741 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4801 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4861 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4921 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4981 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     5041 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     5101 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttgccat tcgccattca
     5161 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagccca
     5221 agctaccatg ataagtaagt aatattaagg tacgggaggt acttggagcg gccgcaataa
     5281 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagtact
//
