LOCUS       sequence-44892-e        5250 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 5250)
  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 5250)
  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: pGL3-321/+120 mut1&2
FEATURES             Location/Qualifiers
     source          1..5250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      514..523
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             520..2172
                     /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(586..604)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2213..2334
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2224..2243)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2278..2297
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2582..2599)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2753..3341)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2833..2852)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3512..4303,4304..4372))
                     /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     4135..4154
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4373..4477)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4504..4959
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4591..4610)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4801..4822
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5090..5138
                     /note="synthetic polyadenylation signal"
     misc_feature    5152..5243
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5192..5211
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcagag gtggagaggc gagagttgcc cgcggctgcc
       61 cctacccctg cccatgtcgc ccacactcca cacgggcccc acagaagcct tcccatcctg
      121 gcttttgttc aacctcggtg ggcggcaagg tgggggcggg acggagtctg cggggcctgc
      181 ggggcccggg cctgcgtcac cgtcactggc ccgcctcccg cccggcctat gggagcccag
      241 gggacacacc ggtcgggggc caggagcgga gattggccag cgggagctcg caccggtgtg
      301 gggcggggag cactgctcct tgtaaggcga gcccccgccg agccggctct tccctcccga
      361 ggcccccacc tcgccaccgc ctcccattgt cccgtcgccc tcgaaactct cccagagacg
      421 ggcgagttgt ccacctgtgc aggtctgaac tgacttcctg tgccaggaga tctgcgatct
      481 aagtaagctt ggcattccgg tactgttggt aaagccacca tggaagacgc caaaaacata
      541 aagaaaggcc cggcgccatt ctatccgctg gaagatggaa ccgctggaga gcaactgcat
      601 aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc
      661 gaggtggaca tcacttacgc tgagtacttc gaaatgtccg ttcggttggc agaagctatg
      721 aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa
      781 ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac
      841 atttataatg aacgtgaatt gctcaacagt atgggcattt cgcagcctac cgtggtgttc
      901 gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa
      961 aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc
     1021 gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtgccaga gtccttcgat
     1081 agggacaaga caattgcact gatcatgaac tcctctggat ctactggtct gcctaaaggt
     1141 gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc atgccagaga tcctattttt
     1201 ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt
     1261 ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga
     1321 tttgaagaag agctgtttct gaggagcctt caggattaca agattcaaag tgcgctgctg
     1381 gtgccaaccc tattctcctt cttcgccaaa agcactctga ttgacaaata cgatttatct
     1441 aatttacacg aaattgcttc tggtggcgct cccctctcta aggaagtcgg ggaagcggtt
     1501 gccaagaggt tccatctgcc aggtatcagg caaggatatg ggctcactga gactacatca
     1561 gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca
     1621 ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcaaaga
     1681 ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa tccggaagcg
     1741 accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac
     1801 gaagacgaac acttcttcat cgttgaccgc ctgaagtctc tgattaagta caaaggctat
     1861 caggtggctc ccgctgaatt ggaatccatc ttgctccaac accccaacat cttcgacgca
     1921 ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg
     1981 gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca
     2041 accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc
     2101 ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag
     2161 atcgccgtgt aattctagag tcggggcggc cggccgcttc gagcagacat gataagatac
     2221 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2281 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2341 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     2401 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtcgaccga tgcccttgag
     2461 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     2521 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt
     2581 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2641 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2701 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2761 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2821 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2881 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2941 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3001 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3061 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3121 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3181 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3241 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3301 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3361 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3421 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3481 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3541 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3601 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3661 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3721 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3781 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3841 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3901 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3961 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4021 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4081 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4141 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4201 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4261 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4321 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4381 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4441 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4501 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4561 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4621 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4681 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4741 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4801 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4861 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4921 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg ccattcgcca ttcaggctgc
     4981 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag cccaagctac
     5041 catgataagt aagtaatatt aaggtacggg aggtacttgg agcggccgca ataaaatatc
     5101 tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac taacatacgc
     5161 tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag
     5221 tgcaggtgcc agaacatttc tctatcgata
//
