LOCUS       sequence-56839-e        6940 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6940)
  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 6940)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pLuc-CDS-m640-2
FEATURES             Location/Qualifiers
     source          1..6940
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        62..253
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(85..105)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      270..405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          489..621
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     604..623
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        666..684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      688..697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             694..1626
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    5068..5116
                     /note="synthetic polyadenylation signal"
     promoter        5149..5253
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5254..5322,5323..6114)
                     /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     complement(5472..5491)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6285..6873
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6774..6793
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
       61 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      421 agcttgattc ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca
      481 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      541 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      601 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
      661 gtacttaata cgactcacta taggctagcc accatggctt ccaaggtgta cgaccccgag
      721 caacgcaaac gcatgatcac tgggcctcag tggtgggctc gctgcaagca aatgaacgtg
      781 ctggactcct tcatcaacta ctatgattcc gagaagcacg ccgagaacgc cgtgattttt
      841 ctgcatggta acgctgcctc cagctacctg tggaggcacg tcgtgcctca catcgagccc
      901 gtggctagat gcatcatccc tgatctgatc ggaatgggta agtccggcaa gagcgggaat
      961 ggctcatatc gcctcctgga tcactacaag tacctcaccg cttggttcga gctgctgaac
     1021 cttccaaaga aaatcatctt tgtgggccac gactgggggg cttgtctggc ctttcactac
     1081 tcctacgagc accaagacaa gatcaaggcc atcgtccatg ctgagagtgt cgtggacgtg
     1141 atcgagtcct gggacgagtg gcctgacatc gaggaggata tcgccctgat caagagcgaa
     1201 gagggcgaga aaatggtgct tgagaataac ttcttcgtcg agaccatgct cccaagcaag
     1261 atcatgcgga aactggagcc tgaggagttc gctgcctacc tggagccatt caaggagaag
     1321 ggcgaggtta gacggcctac cctctcctgg cctcgcgaga tccctctcgt taagggaggc
     1381 aagcccgacg tcgtccagat tgtccgcaac tacaacgcct accttcgggc cagcgacgat
     1441 ctgcctaaga tgttcatcga gtccgaccct gggttctttt ccaacgctat tgtcgaggga
     1501 gctaagaagt tccctaacac cgagttcgtg aaggtgaagg gcctccactt cagccaggag
     1561 gacgctccag atgaaatggg taagtacatc aagagcttcg tggagcgcgt gctgaagaac
     1621 gagcagtcta ggcgatcgct cgagcccggg aattcggtcg acatgaagca gccgatcatg
     1681 gcggatggcc ccaggtgtaa gcgcagaaag caggcgaacc cgcggcgcaa taacgttaca
     1741 aattataata ctgtggtaga aacaaattca gattcagatg atgaagacaa actgcatatt
     1801 gtggaagaag aaagtgttac agatgcagct gactgtgaag gtgtaccaga ggatgacctg
     1861 ccaacagacc agacagtgtt accagggagg agcagtgaaa gagaagggaa tgctaagaac
     1921 tgctgggagg atgacagaaa ggaagggcaa gaaatcctgg ggcctgaagc tcaggcagat
     1981 gaagcaggat gtacagtaaa agatgatgaa tgcgagtcag atgcagaaaa tgagcaaaac
     2041 catgatccta atgttgaaga gtttctacaa caacaagaca ctgctgtcat ttttcctgag
     2101 gcacctgaag aggaccagag gcagggcaca ccagaagcca gtggtcatga tgaaaatgga
     2161 acaccagatg cattttcaca attactcacc tgtccatatt gtgatagagg ctataaacgc
     2221 tttacctctc tgaaagaaca cattaaatat cgtcatgaaa agaatgaaga taactttagt
     2281 tgctccctgt gcagttacac ctttgcatac agaacccaac ttgaacgtca catgacatca
     2341 cataaatcag gaagagatca aagacatgtg acgcagtctg ggtgtaatcg taaattcaaa
     2401 tgcactgagt gtggaaaagc tttcaaatac aaacatcacc taaaagagca cttaagaatt
     2461 cacagtggag agaagccata tgaatgccca aactgcaaga aacgcttttc ccattctggc
     2521 tcctatagct cacacataag cagtaagaaa tgtatcagct tgatacctgt gaatgggcga
     2581 ccaagaacag gactcaagac atctcagtgt tcttcaccgt ctctttcagc atcaccaggc
     2641 agtcccacac gaccacagat acggcaaaag atagagaata aaccccttca agaacaactt
     2701 tctgttaacc aaattaaaac tgaacctgtg gattatgaat tcaaacccat agtggttgct
     2761 tcaggaatca actgttcaac ccctttacaa aatggggttt tcactggtgg tggcccatta
     2821 caggcaacca gttctcctca gggcatggtg caagctgttg ttctgccaac agttggtttg
     2881 gtgtctccca taagtatcaa tttaagtgat attcagaatg tacttaaagt ggcggtagat
     2941 ggtaatgtaa taaggcaagt gttggagaat aatcaagcca atcttgcatc caaagaacaa
     3001 gaaacaatca atgcttcacc catacaacaa ggtggccatt ctgttatttc agccatcagt
     3061 cttcctttgg ttgatcaaga tggaacaacc aaaattatca tcaactacag tcttgagcag
     3121 cctagccaac ttcaagttgt tcctcaaaat ttaaaaaaag aaaatccagt cgctacaaac
     3181 agttgtaaaa gtgaaaagtt accagaagat cttactgtta agtctgagaa ggacaaaagc
     3241 tttgaagggg gggttaacga ctcaacatgt cttctgtgtg atgattgtcc aggagatatt
     3301 aatgcacttc cagaattaaa gcactatgac ctaaagcagc ctactcagcc tcctccactc
     3361 cctgcagcag aagctgagaa gcctgagtcc tctgtttcat cagctactgg agatggcaat
     3421 ttgtctccta gtcagccacc tttaaagaac ctcttgtctc tcctaaaagc atattatgct
     3481 ttgaatgcac aaccaagtgc agaagagctc tcaaaaattg ctgattcagt aaacctacca
     3541 ctggatgtag taaaaaagtg gtttgaaaag atgcaagctg gacagatttc agtgcagtct
     3601 tctgaaccat cttctcctga accaggcaaa gtaaatatcc ctgccaagaa caatgatcag
     3661 cctcaatctg caaatgcaaa tgaaccccag gacagcacag taaatctaca aagtcctttg
     3721 aagatgacta actccccagt tttaccagtg ggatcaacca ccaatggttc cagaagtagt
     3781 acaccatccc catcacctct aaacctttcc tcatccagaa atacacaggg ttacttgtac
     3841 acagctgagg gtgcacaaga agagccacaa gtagaacctc ttgatctttc actaccaaag
     3901 caacagggag aattattaga aaggtcaact atcactagtg tttaccagaa cagtgtttat
     3961 tctgtccagg aagaaccctt gaacttgtct tgcgcaaaaa aggagccaca aaaggacagt
     4021 tgtgttacag actcagaacc agttgtaaat gtaatcccac caagtgccaa ccccataaat
     4081 atcgctatac ctacagtcac tgcccagtta cccacaatcg tggccattgc tgaccagaac
     4141 agtgttccat gcttaagagc gctagctgcc aataagcaaa cgattctgat tccccaggtg
     4201 gcatacacct actcaactac ggtcagccct gcagtccaag aaccaccctt gaaagtgatc
     4261 cagccaaatg gaaatcagga tgaaagacaa gatactagct cagaaggagt atcaaatgta
     4321 gaggatcaga atgactctga ttctacaccg cccaaaaaga aaatgcggaa gacagaaaat
     4381 ggaatgtatg cttgtgattt gtgtgacaag atattccaaa agagtagttc attattgaga
     4441 cataaatatg aacacacagg taaaagacct catgagtgtg gaatctgtaa aaaggcattt
     4501 aaacacaaac atcatttgat tgaacacatg cgattacatt ctggagaaaa gccctatcaa
     4561 tgtgacaaat gtggaaagcg cttctcacac tctgggtctt attctcaaca catgaatcat
     4621 cgctactcct actgtaagag agaagcggaa gaacgtgaca gcacagagca ggaagaggca
     4681 gggcctgaaa tcctctcgaa tgagcacgtg ggtgccaggg cgtctccctc acagggcgac
     4741 tcggacgaga gagagagttt gacaagggaa gaggatgaag acagtgaaaa agaggaagag
     4801 gaggaggata aagagatgga agaattgcag gaagaaaaag aatgtgaaaa accacaaggg
     4861 gatgaggaag aggaggagga ggaggaagaa gtggaagaag aagaggtaga agaggcagag
     4921 aatgagggag aagaagcaaa aactgaaggt ctgatgaagg atgacagggc tgaaagtcaa
     4981 gcaagcagct taggacaaaa agtaggcgag agtagtgagc aagtgtctga agaaaagaca
     5041 aatgaagcct aagcggccgc tggccgcaat aaaatatctt tattttcatt acatctgtgt
     5101 gttggttttt tgtgtgagga tccaggtggc acttttcggg gaaatgtgcg cggaacccct
     5161 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5221 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5281 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5341 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5401 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5461 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5521 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5581 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5641 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5701 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5761 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5821 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5881 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5941 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6001 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6061 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6121 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6181 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6241 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6301 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6361 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6421 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6481 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6541 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6601 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6661 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6721 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6781 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6841 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6901 ttcctggcct tttgctggcc ttttgctcac atggctcgac
//
