LOCUS       sequence-44888-e        4935 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 4935)
  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 4935)
  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: pGL3-110/+19
FEATURES             Location/Qualifiers
     source          1..4935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      200..209
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             206..1858
                     /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(272..290)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1899..2020
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1910..1929)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1964..1983
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2268..2285)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2439..3027)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2519..2538)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3198..3989,3990..4058))
                     /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     3821..3840
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4059..4163)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4190..4645
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4277..4296)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4487..4508
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4776..4824
                     /note="synthetic polyadenylation signal"
     misc_feature    4838..4929
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4878..4897
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcgggg acacaccggt cgggggccag gagcggagat
       61 tggccagcgg gagctcgcac ctgtgtgggg cggggagcac tgctccttgt aaggcgagcc
      121 cccgccgagc cggctcttcc ctcccgaggc cccagatctg cgatctaagt aagcttggca
      181 ttccggtact gttggtaaag ccaccatgga agacgccaaa aacataaaga aaggcccggc
      241 gccattctat ccgctggaag atggaaccgc tggagagcaa ctgcataagg ctatgaagag
      301 atacgccctg gttcctggaa caattgcttt tacagatgca catatcgagg tggacatcac
      361 ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac gatatgggct
      421 gaatacaaat cacagaatcg tcgtatgcag tgaaaactct cttcaattct ttatgccggt
      481 gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg aacgacattt ataatgaacg
      541 tgaattgctc aacagtatgg gcatttcgca gcctaccgtg gtgttcgttt ccaaaaaggg
      601 gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc atccaaaaaa ttattatcat
      661 ggattctaaa acggattacc agggatttca gtcgatgtac acgttcgtca catctcatct
      721 acctcccggt tttaatgaat acgattttgt gccagagtcc ttcgataggg acaagacaat
      781 tgcactgatc atgaactcct ctggatctac tggtctgcct aaaggtgtcg ctctgcctca
      841 tagaactgcc tgcgtgagat tctcgcatgc cagagatcct atttttggca atcaaatcat
      901 tccggatact gcgattttaa gtgttgttcc attccatcac ggttttggaa tgtttactac
      961 actcggatat ttgatatgtg gatttcgagt cgtcttaatg tatagatttg aagaagagct
     1021 gtttctgagg agccttcagg attacaagat tcaaagtgcg ctgctggtgc caaccctatt
     1081 ctccttcttc gccaaaagca ctctgattga caaatacgat ttatctaatt tacacgaaat
     1141 tgcttctggt ggcgctcccc tctctaagga agtcggggaa gcggttgcca agaggttcca
     1201 tctgccaggt atcaggcaag gatatgggct cactgagact acatcagcta ttctgattac
     1261 acccgagggg gatgataaac cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa
     1321 ggttgtggat ctggataccg ggaaaacgct gggcgttaat caaagaggcg aactgtgtgt
     1381 gagaggtcct atgattatgt ccggttatgt aaacaatccg gaagcgacca acgccttgat
     1441 tgacaaggat ggatggctac attctggaga catagcttac tgggacgaag acgaacactt
     1501 cttcatcgtt gaccgcctga agtctctgat taagtacaaa ggctatcagg tggctcccgc
     1561 tgaattggaa tccatcttgc tccaacaccc caacatcttc gacgcaggtg tcgcaggtct
     1621 tcccgacgat gacgccggtg aacttcccgc cgccgttgtt gttttggagc acggaaagac
     1681 gatgacggaa aaagagatcg tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt
     1741 gcgcggagga gttgtgtttg tggacgaagt accgaaaggt cttaccggaa aactcgacgc
     1801 aagaaaaatc agagagatcc tcataaaggc caagaagggc ggaaagatcg ccgtgtaatt
     1861 ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg
     1921 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     1981 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2041 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     2101 caaatgtggt aaaatcgata aggatccgtc gaccgatgcc cttgagagcc ttcaacccag
     2161 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     2221 ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac
     2281 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2341 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2401 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2461 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2521 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2581 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2641 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2701 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2761 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2821 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     2881 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2941 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3001 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3061 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3121 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3181 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3241 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3301 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3361 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3421 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3481 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3541 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3601 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3661 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3721 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3781 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3841 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3901 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3961 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4021 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4081 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4141 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
     4201 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4261 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4321 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4381 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4441 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4501 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4561 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4621 aacaaaatat taacgcttac aatttgccat tcgccattca ggctgcgcaa ctgttgggaa
     4681 gggcgatcgg tgcgggcctc ttcgctatta cgccagccca agctaccatg ataagtaagt
     4741 aatattaagg tacgggaggt acttggagcg gccgcaataa aatatcttta ttttcattac
     4801 atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca
     4861 aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa
     4921 catttctcta tcgat
//
