LOCUS       sequence-44887-e        4986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4986)
  AUTHORS   Bracken CP, Gregory PA, Kolesnikoff N, Bert AG, Wang J, Shannon MF, 
            Goodall GJ
  TITLE     A double-negative feedback loop between ZEB1-SIP1 and the 
            microRNA-200 family regulates epithelial-mesenchymal transition.
  JOURNAL   Cancer Res. 2008 Oct 1;68(19):7846-54.
  PUBMED    18829540
REFERENCE   2  (bases 1 to 4986)
  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: "Cancer 
            Res."; date: "2008-10-1"; volume: "68(19)"; pages: "7846-54"
COMMENT     Sequence Label: pGL3-158/+19
FEATURES             Location/Qualifiers
     source          1..4986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      250..259
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             256..1908
                     /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(322..340)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1949..2070
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1960..1979)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2014..2033
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2318..2335)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2489..3077)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2569..2588)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3248..4039,4040..4108))
                     /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     3871..3890
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4109..4213)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4240..4695
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4327..4346)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4537..4558
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4826..4874
                     /note="synthetic polyadenylation signal"
     misc_feature    4888..4979
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4928..4947
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcgcct gcgtcaccgt cactggcccg cctcccgccc
       61 ggcctatggg agcccagggg acacacctgt cgggggccag gagcggagat tggccagcgg
      121 gagctcgcac ctgtgtgggg cggggagcac tgctccttgt aaggcgagcc cccgccgagc
      181 cggctcttcc ctcccgaggc cccagatctg cgatctaagt aagcttggca ttccggtact
      241 gttggtaaag ccaccatgga agacgccaaa aacataaaga aaggcccggc gccattctat
      301 ccgctggaag atggaaccgc tggagagcaa ctgcataagg ctatgaagag atacgccctg
      361 gttcctggaa caattgcttt tacagatgca catatcgagg tggacatcac ttacgctgag
      421 tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac gatatgggct gaatacaaat
      481 cacagaatcg tcgtatgcag tgaaaactct cttcaattct ttatgccggt gttgggcgcg
      541 ttatttatcg gagttgcagt tgcgcccgcg aacgacattt ataatgaacg tgaattgctc
      601 aacagtatgg gcatttcgca gcctaccgtg gtgttcgttt ccaaaaaggg gttgcaaaaa
      661 attttgaacg tgcaaaaaaa gctcccaatc atccaaaaaa ttattatcat ggattctaaa
      721 acggattacc agggatttca gtcgatgtac acgttcgtca catctcatct acctcccggt
      781 tttaatgaat acgattttgt gccagagtcc ttcgataggg acaagacaat tgcactgatc
      841 atgaactcct ctggatctac tggtctgcct aaaggtgtcg ctctgcctca tagaactgcc
      901 tgcgtgagat tctcgcatgc cagagatcct atttttggca atcaaatcat tccggatact
      961 gcgattttaa gtgttgttcc attccatcac ggttttggaa tgtttactac actcggatat
     1021 ttgatatgtg gatttcgagt cgtcttaatg tatagatttg aagaagagct gtttctgagg
     1081 agccttcagg attacaagat tcaaagtgcg ctgctggtgc caaccctatt ctccttcttc
     1141 gccaaaagca ctctgattga caaatacgat ttatctaatt tacacgaaat tgcttctggt
     1201 ggcgctcccc tctctaagga agtcggggaa gcggttgcca agaggttcca tctgccaggt
     1261 atcaggcaag gatatgggct cactgagact acatcagcta ttctgattac acccgagggg
     1321 gatgataaac cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat
     1381 ctggataccg ggaaaacgct gggcgttaat caaagaggcg aactgtgtgt gagaggtcct
     1441 atgattatgt ccggttatgt aaacaatccg gaagcgacca acgccttgat tgacaaggat
     1501 ggatggctac attctggaga catagcttac tgggacgaag acgaacactt cttcatcgtt
     1561 gaccgcctga agtctctgat taagtacaaa ggctatcagg tggctcccgc tgaattggaa
     1621 tccatcttgc tccaacaccc caacatcttc gacgcaggtg tcgcaggtct tcccgacgat
     1681 gacgccggtg aacttcccgc cgccgttgtt gttttggagc acggaaagac gatgacggaa
     1741 aaagagatcg tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga
     1801 gttgtgtttg tggacgaagt accgaaaggt cttaccggaa aactcgacgc aagaaaaatc
     1861 agagagatcc tcataaaggc caagaagggc ggaaagatcg ccgtgtaatt ctagagtcgg
     1921 ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     1981 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2041 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2101 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     2161 aaaatcgata aggatccgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt
     2221 ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca
     2281 actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2341 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2401 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2461 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2521 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2581 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2641 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2701 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2761 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2821 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2881 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2941 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3001 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3061 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3121 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3181 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3241 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3301 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3361 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3421 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3481 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3541 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3601 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3661 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3721 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3781 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3841 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3901 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3961 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4021 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4081 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4141 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4201 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     4261 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4321 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4381 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4441 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4501 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4561 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4621 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4681 taacgcttac aatttgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     4741 tgcgggcctc ttcgctatta cgccagccca agctaccatg ataagtaagt aatattaagg
     4801 tacgggaggt acttggagcg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     4861 tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca
     4921 aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa catttctcta
     4981 tcgata
//
