LOCUS       sequence-56825-e        5077 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 5077)
  AUTHORS   Mazda M, Nishi K, Naito Y, Ui-Tei K
  TITLE     E-cadherin is transcriptionally activated via suppression of ZEB1 
            transcriptional repressor by small RNA-mediated gene silencing.
  JOURNAL   PLoS One. 2011;6(12):e28688. doi: 10.1371/journal.pone.0028688. Epub
            2011 Dec 21.
  PUBMED    22205962
REFERENCE   2  (bases 1 to 5077)
  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: "PLoS One.";
            date: "2011"; volume: "6(12):e28688. doi"; pages: " 
            10.1371/journal.pone.0028688. Epub 2011 Dec 21"
COMMENT     Sequence Label: proE-cad178-Luc-mEbox
FEATURES             Location/Qualifiers
     source          1..5077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      341..350
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             347..1999
                     /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(413..431)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2040..2161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2051..2070)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2105..2124
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2409..2426)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2580..3168)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2660..2679)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3339..4130,4131..4199))
                     /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     3962..3981
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4200..4304)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4331..4786
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4418..4437)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4628..4649
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4917..4965
                     /note="synthetic polyadenylation signal"
     misc_feature    4979..5070
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5019..5038
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc ttccaggcta gagggtcacc
       61 gcgtctatgc gaggccgggt gggcgggccg tcagctccgc cctggggagg ggtccgcgct
      121 gctgattggc tgtggccgga aggtaaaccc tcagccaatc agcggtacgg ggggcggtgc
      181 ctccggggct aacctagctg cagccacgca ccccctctca gtggcgtcgg aactgcaaag
      241 cacctgtgag cttgcggaag tcagttcaga ctccagcccg ctccagcccg gcccgacccg
      301 accgcacccg gaagcttggc attccggtac tgttggtaaa gccaccatgg aagacgccaa
      361 aaacataaag aaaggcccgg cgccattcta tccgctggaa gatggaaccg ctggagagca
      421 actgcataag gctatgaaga gatacgccct ggttcctgga acaattgctt ttacagatgc
      481 acatatcgag gtggacatca cttacgctga gtacttcgaa atgtccgttc ggttggcaga
      541 agctatgaaa cgatatgggc tgaatacaaa tcacagaatc gtcgtatgca gtgaaaactc
      601 tcttcaattc tttatgccgg tgttgggcgc gttatttatc ggagttgcag ttgcgcccgc
      661 gaacgacatt tataatgaac gtgaattgct caacagtatg ggcatttcgc agcctaccgt
      721 ggtgttcgtt tccaaaaagg ggttgcaaaa aattttgaac gtgcaaaaaa agctcccaat
      781 catccaaaaa attattatca tggattctaa aacggattac cagggatttc agtcgatgta
      841 cacgttcgtc acatctcatc tacctcccgg ttttaatgaa tacgattttg tgccagagtc
      901 cttcgatagg gacaagacaa ttgcactgat catgaactcc tctggatcta ctggtctgcc
      961 taaaggtgtc gctctgcctc atagaactgc ctgcgtgaga ttctcgcatg ccagagatcc
     1021 tatttttggc aatcaaatca ttccggatac tgcgatttta agtgttgttc cattccatca
     1081 cggttttgga atgtttacta cactcggata tttgatatgt ggatttcgag tcgtcttaat
     1141 gtatagattt gaagaagagc tgtttctgag gagccttcag gattacaaga ttcaaagtgc
     1201 gctgctggtg ccaaccctat tctccttctt cgccaaaagc actctgattg acaaatacga
     1261 tttatctaat ttacacgaaa ttgcttctgg tggcgctccc ctctctaagg aagtcgggga
     1321 agcggttgcc aagaggttcc atctgccagg tatcaggcaa ggatatgggc tcactgagac
     1381 tacatcagct attctgatta cacccgaggg ggatgataaa ccgggcgcgg tcggtaaagt
     1441 tgttccattt tttgaagcga aggttgtgga tctggatacc gggaaaacgc tgggcgttaa
     1501 tcaaagaggc gaactgtgtg tgagaggtcc tatgattatg tccggttatg taaacaatcc
     1561 ggaagcgacc aacgccttga ttgacaagga tggatggcta cattctggag acatagctta
     1621 ctgggacgaa gacgaacact tcttcatcgt tgaccgcctg aagtctctga ttaagtacaa
     1681 aggctatcag gtggctcccg ctgaattgga atccatcttg ctccaacacc ccaacatctt
     1741 cgacgcaggt gtcgcaggtc ttcccgacga tgacgccggt gaacttcccg ccgccgttgt
     1801 tgttttggag cacggaaaga cgatgacgga aaaagagatc gtggattacg tcgccagtca
     1861 agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt gtggacgaag taccgaaagg
     1921 tcttaccgga aaactcgacg caagaaaaat cagagagatc ctcataaagg ccaagaaggg
     1981 cggaaagatc gccgtgtaat tctagagtcg gggcggccgg ccgcttcgag cagacatgat
     2041 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     2101 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     2161 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     2221 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatccgt cgaccgatgc
     2281 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     2341 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     2401 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2461 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2521 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2581 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2641 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2701 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2761 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2821 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2881 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2941 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3001 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3061 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3121 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3181 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3241 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3301 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3361 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3421 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3481 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3541 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3601 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3661 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3721 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3781 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3841 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3901 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3961 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4021 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4081 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4141 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4201 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4261 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4321 gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4381 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     4441 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4501 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     4561 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4621 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     4681 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     4741 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttgcca ttcgccattc
     4801 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagccc
     4861 aagctaccat gataagtaag taatattaag gtacgggagg tacttggagc ggccgcaata
     4921 aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagtactaa
     4981 catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca
     5041 gtgcaagtgc aggtgccaga acatttctct atcgata
//
