LOCUS       sequence-44885-e        6503 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 6503)
  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 6503)
  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: pGL3-1574/+120
FEATURES             Location/Qualifiers
     source          1..6503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1767..1776
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1773..3425
                     /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(1839..1857)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3466..3587
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3477..3496)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3531..3550
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3835..3852)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4006..4594)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4086..4105)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4765..5556,5557..5625))
                     /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     5388..5407
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5626..5730)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5757..6212
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5844..5863)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6054..6075
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6343..6391
                     /note="synthetic polyadenylation signal"
     misc_feature    6405..6496
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6445..6464
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagccatt cccggacagt cacagccgtg gaacaacgca
       61 gcgtgtggac acagccccag ggaagcttga ctctgcctgg tccccatggc gtgcggtggc
      121 tcaccagccg cccccatcgc tccatgaaat ctgactacac ctgtcccaaa aatcaaccag
      181 ctcccaggca ggtcccgccg tgcagggggc caagggccac acctgggacc agtgagtgag
      241 gcggcgtgcg ggtagtgggg gtgtgagccc gagacggggg tccagccttc tcccacttgg
      301 atgccaacag cagactctgg atttggaggt gtgggcggtg ccttagtctc cccagaagac
      361 aagagcccac ccgccgggtg tgtcctgacc ccacccccac ctgccccaca cagacacctg
      421 agtgggcatc agagtgggcc gggccctgac tgtccggggg agggactgtt ggggcaaggg
      481 cagtgggtca gaggtcaccg gtgcccccag gaccagtttc cagcgagaag agagccagga
      541 gccccgggca caggcccacg gtgatgaggc tcctgaccga gctcccagag ctcattatgg
      601 aaaaccgtgg ggtccgctgg gacctctctg ctggtcgggc taattacaga ccgtttgtcg
      661 tttcattatg ggctcttcct ctgggagatg cgagaagcaa acactttgcc atgggagggg
      721 cgcttcccgg accgggcggg ggccctgccc tccccacccc cagacccctg gtcccccagg
      781 ccttccaagc tcccaaagaa aacttgaacc cccactccag gggctccaaa gtaacctcca
      841 gagcccagat ggggatgttc cgggagctgc tggggcctct cagctgtccc tgcctcgcag
      901 ggaggctcag gaaggcaccc acacccctcc ctcattcccc cagctgtggg tctgtggggt
      961 ctcggagcct cttgcccatt ggggtgggac tgggggcacg tagctcctcc gagcccctgg
     1021 cgaggagacc gatcctgaga cccacctggg cctacggggg gtcccttcat tgctccaaaa
     1081 ttaagggctg cgggcctgga gcaaagacag gcaggtatgg cccctgccct ctgtccatga
     1141 gaggcctcag ccccatggga cagggcacca ccgctgcagg gaggcggcgg gcaggttgtg
     1201 acccctcacc ctggggccac cctacccgct tctaaggcag cctgggaagg agccctgcgg
     1261 agctgggggg tggggaggca gaggtggaga ggcgagagtt gcccgcggct gcccctaccc
     1321 ctgcccatgt cgcccacact ccacacgggc cccacagaag ccttcccatc ctggcttttg
     1381 ttcaacctcg gtgggcggca aggtgggggc gggacggagt ctgcggggcc tgcggggccc
     1441 gggcctgcgt caccgtcact ggcccgcctc ccgcccggcc tatgggagcc caggggacac
     1501 acctgtcggg ggccaggagc ggagattggc cagcgggagc tcgcacctgt gtggggcggg
     1561 gagcactgct ccttgtaagg cgagcccccg ccgagccggc tcttccctcc cgaggccccc
     1621 acctcgccac cgcctcccat tgtcccgtcg ccctcgaaac tctcccagag acgggcgagt
     1681 tgtccacctg tgcaggtctg aactgacttc ctgtgccagg agatctgcga tctaagtaag
     1741 cttggcattc cggtactgtt ggtaaagcca ccatggaaga cgccaaaaac ataaagaaag
     1801 gcccggcgcc attctatccg ctggaagatg gaaccgctgg agagcaactg cataaggcta
     1861 tgaagagata cgccctggtt cctggaacaa ttgcttttac agatgcacat atcgaggtgg
     1921 acatcactta cgctgagtac ttcgaaatgt ccgttcggtt ggcagaagct atgaaacgat
     1981 atgggctgaa tacaaatcac agaatcgtcg tatgcagtga aaactctctt caattcttta
     2041 tgccggtgtt gggcgcgtta tttatcggag ttgcagttgc gcccgcgaac gacatttata
     2101 atgaacgtga attgctcaac agtatgggca tttcgcagcc taccgtggtg ttcgtttcca
     2161 aaaaggggtt gcaaaaaatt ttgaacgtgc aaaaaaagct cccaatcatc caaaaaatta
     2221 ttatcatgga ttctaaaacg gattaccagg gatttcagtc gatgtacacg ttcgtcacat
     2281 ctcatctacc tcccggtttt aatgaatacg attttgtgcc agagtccttc gatagggaca
     2341 agacaattgc actgatcatg aactcctctg gatctactgg tctgcctaaa ggtgtcgctc
     2401 tgcctcatag aactgcctgc gtgagattct cgcatgccag agatcctatt tttggcaatc
     2461 aaatcattcc ggatactgcg attttaagtg ttgttccatt ccatcacggt tttggaatgt
     2521 ttactacact cggatatttg atatgtggat ttcgagtcgt cttaatgtat agatttgaag
     2581 aagagctgtt tctgaggagc cttcaggatt acaagattca aagtgcgctg ctggtgccaa
     2641 ccctattctc cttcttcgcc aaaagcactc tgattgacaa atacgattta tctaatttac
     2701 acgaaattgc ttctggtggc gctcccctct ctaaggaagt cggggaagcg gttgccaaga
     2761 ggttccatct gccaggtatc aggcaaggat atgggctcac tgagactaca tcagctattc
     2821 tgattacacc cgagggggat gataaaccgg gcgcggtcgg taaagttgtt ccattttttg
     2881 aagcgaaggt tgtggatctg gataccggga aaacgctggg cgttaatcaa agaggcgaac
     2941 tgtgtgtgag aggtcctatg attatgtccg gttatgtaaa caatccggaa gcgaccaacg
     3001 ccttgattga caaggatgga tggctacatt ctggagacat agcttactgg gacgaagacg
     3061 aacacttctt catcgttgac cgcctgaagt ctctgattaa gtacaaaggc tatcaggtgg
     3121 ctcccgctga attggaatcc atcttgctcc aacaccccaa catcttcgac gcaggtgtcg
     3181 caggtcttcc cgacgatgac gccggtgaac ttcccgccgc cgttgttgtt ttggagcacg
     3241 gaaagacgat gacggaaaaa gagatcgtgg attacgtcgc cagtcaagta acaaccgcga
     3301 aaaagttgcg cggaggagtt gtgtttgtgg acgaagtacc gaaaggtctt accggaaaac
     3361 tcgacgcaag aaaaatcaga gagatcctca taaaggccaa gaagggcgga aagatcgccg
     3421 tgtaattcta gagtcggggc ggccggccgc ttcgagcaga catgataaga tacattgatg
     3481 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     3541 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     3601 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     3661 acctctacaa atgtggtaaa atcgataagg atccgtcgac cgatgccctt gagagccttc
     3721 aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact
     3781 gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctcttccg cttcctcgct
     3841 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3901 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3961 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4021 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4081 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4141 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4201 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4261 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4321 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4381 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4441 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4501 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4561 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4621 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4681 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4741 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4801 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4861 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4921 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4981 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5041 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5101 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5161 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5221 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5281 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5341 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5401 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5461 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5521 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5581 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5641 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5701 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc
     5761 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5821 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5881 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5941 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     6001 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     6061 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     6121 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     6181 gaattttaac aaaatattaa cgcttacaat ttgccattcg ccattcaggc tgcgcaactg
     6241 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagcccaagc taccatgata
     6301 agtaagtaat attaaggtac gggaggtact tggagcggcc gcaataaaat atctttattt
     6361 tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag tactaacata cgctctccat
     6421 caaaacaaaa cgaaacaaaa caaactagca aaataggctg tccccagtgc aagtgcaggt
     6481 gccagaacat ttctctatcg ata
//
