LOCUS       sequence-56824-e        5569 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5569)
  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 5569)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-cad670-Luc
FEATURES             Location/Qualifiers
     source          1..5569
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      833..842
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             839..2491
                     /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(905..923)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2532..2653
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2543..2562)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2597..2616
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2901..2918)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3072..3660)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3152..3171)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3831..4622,4623..4691))
                     /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     4454..4473
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4692..4796)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4823..5278
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4910..4929)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5120..5141
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5409..5457
                     /note="synthetic polyadenylation signal"
     misc_feature    5471..5562
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5511..5530
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc tggctgctag ctcagtggct
       61 catggctcac acctgaaatc ctagcacttt gggaggccaa ggcaggagga tcgcttcagc
      121 ccaggagttc gagaccaggc tgggcaatac agggagacac agcgccccca ctgcccctgt
      181 ccgccccgac ttgtctctct acaaaaaggc aaaagaaaaa aaaattagcc ttgcgtggtg
      241 gtgtgcacct gtactcccag ctactagaga ggctggggcc agaggaccgc ttgagcccag
      301 gagttcgagg ctgcagtgag ctgtgatcgc accactgcac tccagcttgg gtgaaagagt
      361 gagaccccat ctccaaaacg aacaaacaaa aaatcccaaa aaacaaaaga actcagccaa
      421 gtgtaaaagc cctttctgat cccaggtctt agtgagccac cggcggggct gggattcgaa
      481 cccagtggaa tcagaaccgt gcaggtccca taacccacct agaccctagc aactccaggc
      541 tagagggtca ccgcgtctat gcgaggccgg gtgggcgggc cgtcagctcc gccctgggga
      601 ggggtccgcg ctgctgattg gctgtggccg gcaggtgaac cctcagccaa tcagcggtac
      661 ggggggcggt gcctccgggg ctcacctggc tgcagccacg caccccctct cagtggcgtc
      721 ggaactgcaa agcacctgtg agcttgcgga agtcagttca gactccagcc cgctccagcc
      781 cggcccgacc cgaccgcacc cggaagcttg gcattccggt actgttggta aagccaccat
      841 ggaagacgcc aaaaacataa agaaaggccc ggcgccattc tatccgctgg aagatggaac
      901 cgctggagag caactgcata aggctatgaa gagatacgcc ctggttcctg gaacaattgc
      961 ttttacagat gcacatatcg aggtggacat cacttacgct gagtacttcg aaatgtccgt
     1021 tcggttggca gaagctatga aacgatatgg gctgaataca aatcacagaa tcgtcgtatg
     1081 cagtgaaaac tctcttcaat tctttatgcc ggtgttgggc gcgttattta tcggagttgc
     1141 agttgcgccc gcgaacgaca tttataatga acgtgaattg ctcaacagta tgggcatttc
     1201 gcagcctacc gtggtgttcg tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa
     1261 aaagctccca atcatccaaa aaattattat catggattct aaaacggatt accagggatt
     1321 tcagtcgatg tacacgttcg tcacatctca tctacctccc ggttttaatg aatacgattt
     1381 tgtgccagag tccttcgata gggacaagac aattgcactg atcatgaact cctctggatc
     1441 tactggtctg cctaaaggtg tcgctctgcc tcatagaact gcctgcgtga gattctcgca
     1501 tgccagagat cctatttttg gcaatcaaat cattccggat actgcgattt taagtgttgt
     1561 tccattccat cacggttttg gaatgtttac tacactcgga tatttgatat gtggatttcg
     1621 agtcgtctta atgtatagat ttgaagaaga gctgtttctg aggagccttc aggattacaa
     1681 gattcaaagt gcgctgctgg tgccaaccct attctccttc ttcgccaaaa gcactctgat
     1741 tgacaaatac gatttatcta atttacacga aattgcttct ggtggcgctc ccctctctaa
     1801 ggaagtcggg gaagcggttg ccaagaggtt ccatctgcca ggtatcaggc aaggatatgg
     1861 gctcactgag actacatcag ctattctgat tacacccgag ggggatgata aaccgggcgc
     1921 ggtcggtaaa gttgttccat tttttgaagc gaaggttgtg gatctggata ccgggaaaac
     1981 gctgggcgtt aatcaaagag gcgaactgtg tgtgagaggt cctatgatta tgtccggtta
     2041 tgtaaacaat ccggaagcga ccaacgcctt gattgacaag gatggatggc tacattctgg
     2101 agacatagct tactgggacg aagacgaaca cttcttcatc gttgaccgcc tgaagtctct
     2161 gattaagtac aaaggctatc aggtggctcc cgctgaattg gaatccatct tgctccaaca
     2221 ccccaacatc ttcgacgcag gtgtcgcagg tcttcccgac gatgacgccg gtgaacttcc
     2281 cgccgccgtt gttgttttgg agcacggaaa gacgatgacg gaaaaagaga tcgtggatta
     2341 cgtcgccagt caagtaacaa ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga
     2401 agtaccgaaa ggtcttaccg gaaaactcga cgcaagaaaa atcagagaga tcctcataaa
     2461 ggccaagaag ggcggaaaga tcgccgtgta attctagagt cggggcggcc ggccgcttcg
     2521 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2581 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2641 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     2701 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc
     2761 gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca
     2821 tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc
     2881 cggcagcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2941 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3001 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3061 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3121 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3181 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3241 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3301 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3361 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3421 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3481 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3541 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3601 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3661 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3721 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3781 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3841 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3901 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3961 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4021 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4081 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4141 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4201 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4261 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4321 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4381 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4441 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4501 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4561 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4621 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4681 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4741 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4801 tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4861 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4921 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4981 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     5041 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     5101 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     5161 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     5221 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttgc
     5281 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     5341 ttacgccagc ccaagctacc atgataagta agtaatatta aggtacggga ggtacttgga
     5401 gcggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat
     5461 cgatagtact aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat
     5521 aggctgtccc cagtgcaagt gcaggtgcca gaacatttct ctatcgata
//
