LOCUS       sequence-44889-e        4892 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 4892)
  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 4892)
  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: pGL3-64/+19
FEATURES             Location/Qualifiers
     source          1..4892
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      156..165
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             162..1814
                     /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(228..246)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1855..1976
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1866..1885)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1920..1939
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2224..2241)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2395..2983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2475..2494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3154..3945,3946..4014))
                     /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     3777..3796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4015..4119)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4146..4601
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4233..4252)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4443..4464
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4732..4780
                     /note="synthetic polyadenylation signal"
     misc_feature    4794..4885
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4834..4853
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcgagc tcgcaccggt gtggggcggg gagcactgct
       61 ccttgtaagg cgagcccccg ccgagccggc tcttccctcc cgaggcccca gatctgcgat
      121 ctaagtaagc ttggcattcc ggtactgttg gtaaagccac catggaagac gccaaaaaca
      181 taaagaaagg cccggcgcca ttctatccgc tggaagatgg aaccgctgga gagcaactgc
      241 ataaggctat gaagagatac gccctggttc ctggaacaat tgcttttaca gatgcacata
      301 tcgaggtgga catcacttac gctgagtact tcgaaatgtc cgttcggttg gcagaagcta
      361 tgaaacgata tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa aactctcttc
      421 aattctttat gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg cccgcgaacg
      481 acatttataa tgaacgtgaa ttgctcaaca gtatgggcat ttcgcagcct accgtggtgt
      541 tcgtttccaa aaaggggttg caaaaaattt tgaacgtgca aaaaaagctc ccaatcatcc
      601 aaaaaattat tatcatggat tctaaaacgg attaccaggg atttcagtcg atgtacacgt
      661 tcgtcacatc tcatctacct cccggtttta atgaatacga ttttgtgcca gagtccttcg
      721 atagggacaa gacaattgca ctgatcatga actcctctgg atctactggt ctgcctaaag
      781 gtgtcgctct gcctcataga actgcctgcg tgagattctc gcatgccaga gatcctattt
      841 ttggcaatca aatcattccg gatactgcga ttttaagtgt tgttccattc catcacggtt
      901 ttggaatgtt tactacactc ggatatttga tatgtggatt tcgagtcgtc ttaatgtata
      961 gatttgaaga agagctgttt ctgaggagcc ttcaggatta caagattcaa agtgcgctgc
     1021 tggtgccaac cctattctcc ttcttcgcca aaagcactct gattgacaaa tacgatttat
     1081 ctaatttaca cgaaattgct tctggtggcg ctcccctctc taaggaagtc ggggaagcgg
     1141 ttgccaagag gttccatctg ccaggtatca ggcaaggata tgggctcact gagactacat
     1201 cagctattct gattacaccc gagggggatg ataaaccggg cgcggtcggt aaagttgttc
     1261 cattttttga agcgaaggtt gtggatctgg ataccgggaa aacgctgggc gttaatcaaa
     1321 gaggcgaact gtgtgtgaga ggtcctatga ttatgtccgg ttatgtaaac aatccggaag
     1381 cgaccaacgc cttgattgac aaggatggat ggctacattc tggagacata gcttactggg
     1441 acgaagacga acacttcttc atcgttgacc gcctgaagtc tctgattaag tacaaaggct
     1501 atcaggtggc tcccgctgaa ttggaatcca tcttgctcca acaccccaac atcttcgacg
     1561 caggtgtcgc aggtcttccc gacgatgacg ccggtgaact tcccgccgcc gttgttgttt
     1621 tggagcacgg aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc agtcaagtaa
     1681 caaccgcgaa aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg aaaggtctta
     1741 ccggaaaact cgacgcaaga aaaatcagag agatcctcat aaaggccaag aagggcggaa
     1801 agatcgccgt gtaattctag agtcggggcg gccggccgct tcgagcagac atgataagat
     1861 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     1921 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     1981 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     2041 gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tccgtcgacc gatgcccttg
     2101 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca
     2161 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctcttccgc
     2221 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2281 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2341 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2401 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2461 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2521 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2581 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2641 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2701 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2761 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2821 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2881 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2941 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3001 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3061 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3121 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3181 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3241 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3301 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3361 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3421 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3481 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3541 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3601 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3661 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3721 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3781 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3841 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3901 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3961 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4021 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4081 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4141 acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4201 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4261 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4321 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4381 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4441 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4501 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4561 atttaacgcg aattttaaca aaatattaac gcttacaatt tgccattcgc cattcaggct
     4621 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agcccaagct
     4681 accatgataa gtaagtaata ttaaggtacg ggaggtactt ggagcggccg caataaaata
     4741 tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt actaacatac
     4801 gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca
     4861 agtgcaggtg ccagaacatt tctctatcga ta
//
