LOCUS       sequence-201215-        5820 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  luciferase reporter to assay Zfp521 promoter activation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5820)
  AUTHORS   Yu M, Al-Dallal S, Al-Haj L, Panjwani S, McCartney AS, Edwards SM, 
            Manjunath P, Walker C, Awgulewitsch A, Hentges KE
  TITLE     Transcriptional regulation of the proto-oncogene Zfp521 by SPI1 
            (PU.1) and HOXC13.
  JOURNAL   Genesis. 2016 Oct;54(10):519-533. doi: 10.1002/dvg.22963. Epub 2016 
            Aug 29.
  PUBMED    27506447
REFERENCE   2  (bases 1 to 5820)
  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: "10.1002/dvg.22963";
            journalName: "Genesis"; date: "2016-10"; volume: "54"; issue: "10"; 
            pages: "519-533"
COMMENT     Sequence Label: 1kb Zfp521 promoter reporter
FEATURES             Location/Qualifiers
     source          1..5820
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      14..469
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(101..120)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     311..332
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    600..648
                     /note="synthetic polyadenylation signal"
     misc_feature    662..753
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     702..721
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      1844..1853
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1850..3502
                     /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(1916..1934)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3543..3664
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3554..3573)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3608..3627
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3912..3929)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4083..4671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4163..4182)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4842..5633,5634..5702))
                     /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     5465..5484
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5703..5807)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
       61 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
      121 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
      181 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
      241 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
      301 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
      361 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
      421 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg ccattcgcca
      481 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
      541 cccaagctac catgataagt aagtaatatt aaggtacggg aggtacttgg agcggccgca
      601 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac
      661 taacatacgc tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc
      721 ccagtgcaag tgcaggtgcc agaacatttc tctatcgata ggtaccgagc tcggatccac
      781 tagtaacggc cgccagtgtg ctggaattcg gcttttctta cctcctgctc attcttaaaa
      841 acaaaacaaa atcagaaaca aaaagaaaaa caaagaaaag gcgttatttc tatgccaact
      901 acagagcgtt atgtaaagta ctgaaccaga gccgttttgt ttcaatcacg ttttgacatt
      961 cttcatcaac cactttctct aacgagggaa agaaaaaaaa agtctaaaca acttgccttt
     1021 aataaagatt ccaaatagag agttggcagg tctgctgttg cttcacaaaa agattccccc
     1081 cgactgggcg actgaagcag tgctaaaaaa taaaagtaca acaataccaa aaccaaaaac
     1141 caaaaccaac aacaacaaaa aagtcctttc tgcccccgct gagaaaataa actgtttcca
     1201 ctttttgtat ccattctacc agaaggggga ccttttccac tttgagatgc tcctcggctt
     1261 taatttctgg agaggaagaa aaaaaaaaaa aaaaaaaaaa agcgagcaga agagccctga
     1321 gatccgcgaa gcagcttcta ataacatagg cagcggcgcg ggtgtgtgcg cgagccgtcg
     1381 gcgcgcgggc gggagggcag gcgggcgcga ggggcggcgg caggagcggg cggggcgcgc
     1441 gcaccggcgg gcgggggcgc gcgggggtgg cagcgccgga gcgggcgggg gcgcgcggcc
     1501 gggctctgcg gccggcgagg ctccggatcc cgcgcggcgc gtccggcgcc tccagtccgg
     1561 gtcttggcgg gaccccctcc tccctcccgc tccccgcgct ctcgcctttt aatcatgccc
     1621 ctctgtctgt gtgtgagtgc aggcaggctg acaatgattt cctcagtgat tacgtacaga
     1681 gcgagtccct gcgggttagg ggccccctct ggagccatcc tgatggcttt gggggccttg
     1741 cttccatttt ccattaagcc gaattctgca gatatccatc acactggcgg ccgctcgaga
     1801 tctgcgatct aagtaagctt ggcattccgg tactgttggt aaagccacca tggaagacgc
     1861 caaaaacata aagaaaggcc cggcgccatt ctatccgctg gaagatggaa ccgctggaga
     1921 gcaactgcat aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga
     1981 tgcacatatc gaggtggaca tcacttacgc tgagtacttc gaaatgtccg ttcggttggc
     2041 agaagctatg aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa
     2101 ctctcttcaa ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc
     2161 cgcgaacgac atttataatg aacgtgaatt gctcaacagt atgggcattt cgcagcctac
     2221 cgtggtgttc gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaagctccc
     2281 aatcatccaa aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat
     2341 gtacacgttc gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtgccaga
     2401 gtccttcgat agggacaaga caattgcact gatcatgaac tcctctggat ctactggtct
     2461 gcctaaaggt gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc atgccagaga
     2521 tcctattttt ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca
     2581 tcacggtttt ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt
     2641 aatgtataga tttgaagaag agctgtttct gaggagcctt caggattaca agattcaaag
     2701 tgcgctgctg gtgccaaccc tattctcctt cttcgccaaa agcactctga ttgacaaata
     2761 cgatttatct aatttacacg aaattgcttc tggtggcgct cccctctcta aggaagtcgg
     2821 ggaagcggtt gccaagaggt tccatctgcc aggtatcagg caaggatatg ggctcactga
     2881 gactacatca gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa
     2941 agttgttcca ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt
     3001 taatcaaaga ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa
     3061 tccggaagcg accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc
     3121 ttactgggac gaagacgaac acttcttcat cgttgaccgc ctgaagtctc tgattaagta
     3181 caaaggctat caggtggctc ccgctgaatt ggaatccatc ttgctccaac accccaacat
     3241 cttcgacgca ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt
     3301 tgttgttttg gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag
     3361 tcaagtaaca accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa
     3421 aggtcttacc ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa
     3481 gggcggaaag atcgccgtgt aattctagag tcggggcggc cggccgcttc gagcagacat
     3541 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     3601 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     3661 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     3721 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtcgaccga
     3781 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
     3841 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
     3901 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3961 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4021 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4081 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4141 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4201 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4261 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4321 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4381 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4441 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4501 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4561 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4621 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4681 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4741 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4801 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4861 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4921 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4981 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5041 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5101 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5161 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5221 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5281 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5341 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5401 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5461 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5521 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5581 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5641 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5701 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5761 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
//
