LOCUS       sequence-216993-        8139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter containing PAK4- FL-3'UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8139)
  AUTHORS   Phatak P, Burrows WM, Chesnick IE, Tulapurkar ME, Rao JN, Turner DJ,
            Hamburger AW, Wang JY, Donahue JM
  TITLE     MiR-199a-3p decreases esophageal cancer cell proliferation by 
            targeting p21 activated kinase 4.
  JOURNAL   Oncotarget. 2018 Jun 19;9(47):28391-28407. doi: 
            10.18632/oncotarget.25375. eCollection 2018 Jun 19.
  PUBMED    29983868
REFERENCE   2  (bases 1 to 8139)
  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"; doi: 
            "10.18632/oncotarget.25375"; journalName: "Oncotarget"; date: 
            "2018-06-19- 19"; volume: "9"; issue: "47"; pages: "28391-28407"
COMMENT     Sequence Label: PAK4-FL-3'UTR
FEATURES             Location/Qualifiers
     source          1..8139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        139..496
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        139..330
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(162..182)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      347..482
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      541..550
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             547..1479
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             1531..2322
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1582..1601)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2192..2211
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2386..2434
                     /note="synthetic polyadenylation signal"
     promoter        2590..2694
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2695..2763,2764..3555)
                     /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(2913..2932)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3713..4301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3793..3812)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4417..4700
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     promoter        4757..5256
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4792..4815)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5189..5208
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      5297..5306
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5303..6955
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    7912..8033
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7923..7942)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7977..7996
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
       61 tttcacaccg catacgcgga tctgcgcagc accatggcct gaaataacct ctgaaagagg
      121 aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
      181 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
      241 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
      301 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
      361 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
      421 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
      481 cctaggcttt tgcaaaaagc ttgattcttc tgacacaaca gtctcgaacc aaaggctgga
      541 gccaccatgg cttccaaggt gtacgacccc gagcaacgca aacgcatgat cactgggcct
      601 cagtggtggg ctcgctgcaa gcaaatgaac gtgctggact ccttcatcaa ctactatgat
      661 tccgagaagc acgccgagaa cgccgtgatt tttctgcatg gtaacgctgc ctccagctac
      721 ctgtggaggc acgtcgtgcc tcacatcgag cccgtggcta gatgcatcat ccctgatctg
      781 atcggaatgg gtaagtccgg caagagcggg aatggctcat atcgcctcct ggatcactac
      841 aagtacctca ccgcttggtt cgagctgctg aaccttccaa agaaaatcat ctttgtgggc
      901 cacgactggg gggcttgtct ggcctttcac tactcctacg agcaccaaga caagatcaag
      961 gccatcgtcc atgctgagag tgtcgtggac gtgatcgagt cctgggacga gtggcctgac
     1021 atcgaggagg atatcgccct gatcaagagc gaagagggcg agaaaatggt gcttgagaat
     1081 aacttcttcg tcgagaccat gctcccaagc aagatcatgc ggaaactgga gcctgaggag
     1141 ttcgctgcct acctggagcc attcaaggag aagggcgagg ttagacggcc taccctctcc
     1201 tggcctcgcg agatccctct cgttaaggga ggcaagcccg acgtcgtcca gattgtccgc
     1261 aactacaacg cctaccttcg ggccagcgac gatctgccta agatgttcat cgagtccgac
     1321 cctgggttct tttccaacgc tattgtcgag ggagctaaga agttccctaa caccgagttc
     1381 gtgaaggtga agggcctcca cttcagccag gaggacgctc cagatgaaat gggtaagtac
     1441 atcaagagct tcgtggagcg cgtgctgaag aacgagcaga ccggtggtgg gagcggaggt
     1501 ggcggatcag gtggcggagg ctccggaggg attgaacaag atggattgca cgcaggttct
     1561 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     1621 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     1681 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     1741 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     1801 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     1861 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     1921 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     1981 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     2041 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     2101 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     2161 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     2221 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     2281 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     2341 aaatgaccga ccaagcgacg cccaacctgc catcacgatg gccgcaataa aatatcttta
     2401 ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagcgataag gatcctcttt
     2461 gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg gggtcagcac
     2521 cgtttctgcg gactggcttt ctacgtaatg gtttcttaga cgtcaggtgg cacttttcgg
     2581 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2641 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     2701 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2761 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     2821 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     2881 cgttttccaa tgatgagcac tttcaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     2941 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3001 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     3061 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac tatcggagga
     3121 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3181 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3241 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     3301 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     3361 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     3421 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3481 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3541 attaagcatt ggtaattcga aatgaccgac caagcgacgc ccaaccggta tcagctcact
     3601 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3661 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3721 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3781 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3841 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3901 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3961 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4021 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4081 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4141 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4201 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4261 tttgcaagca gcagattacg cgcagaaaaa aaggatttca agaagatcct ttgatctttt
     4321 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4381 tatcaaaaag gatcttcacc tagatccttt tatagtccgg aaatacagga acgcacgctg
     4441 gatggccctt cgctgggatg gtgaaaccat gaaaaatggc agcttcagtg gattaagtgg
     4501 gggtaatgtg gcctgtaccc tctggttgca taggtattca tacggttaaa atttatcagg
     4561 cgcgattgcg gcagtttttc gggtggtttg ttgccatttt tacctgtctg ctgccgtgat
     4621 cgcgctgaac gcgttttagc ggtgcgtaca attaagggat tatggtaaat ccacttactg
     4681 tctgccctcg tagccatcga gataaaccgc agtactccgg ccacgatgcg tccggcgtag
     4741 aggatcgaga tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct
     4801 ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc
     4861 acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct
     4921 actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga
     4981 caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg
     5041 aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct
     5101 cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg
     5161 ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc
     5221 gcgctgttct cctcttcctc atctccgggc ctttcgacct gcagcccaag cttggcaatc
     5281 cggtactgtt ggtaaagcca ccatggaaga tgccaaaaac attaagaagg gcccagcgcc
     5341 attctaccca ctcgaagacg ggaccgccgg cgagcagctg cacaaagcca tgaagcgcta
     5401 cgccctggtg cccggcacca tcgcctttac cgacgcacat atcgaggtgg acattaccta
     5461 cgccgagtac ttcgagatga gcgttcggct ggcagaagct atgaagcgct atgggctgaa
     5521 tacaaaccat cggatcgtgg tgtgcagcga gaatagcttg cagttcttca tgcccgtgtt
     5581 gggtgccctg ttcatcggtg tggctgtggc cccagctaac gacatctaca acgagcgcga
     5641 gctgctgaac agcatgggca tcagccagcc caccgtcgta ttcgtgagca agaaagggct
     5701 gcaaaagatc ctcaacgtgc aaaagaagct accgatcata caaaagatca tcatcatgga
     5761 tagcaagacc gactaccagg gcttccaaag catgtacacc ttcgtgactt cccatttgcc
     5821 acccggcttc aacgagtacg acttcgtgcc cgagagcttc gaccgggaca aaaccatcgc
     5881 cctgatcatg aacagtagtg gcagtaccgg attgcccaag ggcgtagccc taccgcaccg
     5941 caccgcttgt gtccgattca gtcatgcccg cgaccccatc ttcggcaacc agatcatccc
     6001 cgacaccgct atcctcagcg tggtgccatt tcaccacggc ttcggcatgt tcaccacgct
     6061 gggctacttg atctgcggct ttcgggtcgt gctcatgtac cgcttcgagg aggagctatt
     6121 cttgcgcagc ttgcaagact ataagattca atctgccctg ctggtgccca cactatttag
     6181 cttcttcgct aagagcactc tcatcgacaa gtacgaccta agcaacttgc acgagatcgc
     6241 cagcggcggg gcgccgctca gcaaggaggt aggtgaggcc gtggccaaac gcttccacct
     6301 accaggcatc cgccagggct acggcctgac agaaacaacc agcgccattc tgatcacccc
     6361 cgaaggggac gacaagcctg gcgcagtagg caaggtggtg cccttcttcg aggctaaggt
     6421 ggtggacttg gacaccggta agacactggg tgtgaaccag cgcggcgagc tgtgcgtccg
     6481 tggccccatg atcatgagcg gctacgttaa caaccccgag gctacaaacg ctctcatcga
     6541 caaggacggc tggctgcaca gcggcgacat cgcctactgg gacgaggacg agcacttctt
     6601 catcgtggac cggctgaaga gcctgatcaa atacaagggc taccaggtag ccccagccga
     6661 actggagagc atcctgctgc aacaccccaa catcttcgac gccggggtcg ccggcctgcc
     6721 cgacgacgat gccggcgagc tgcccgccgc agtcgtcgtg ctggaacacg gtaaaaccat
     6781 gaccgagaag gagatcgtgg actatgtggc cagccaggtt acaaccgcca agaagctgcg
     6841 cggtggtgtt gtgttcgtgg acgaggtgcc taaaggactg accggcaagt tggacgcccg
     6901 caagatccgc gagattctca ttaaggccaa gaagggcggc aagatcgccg tgtaattcta
     6961 gttgtttaaa cccttcccct caaccaaaga gccccccggg tcacccccgc cccactgagg
     7021 ccagtagggg gccaggcctc ccactcctcc cagcccggga gatgctccgt gtggcaccac
     7081 cctccttgct gggggtagat gagaccctac tactgaactc cagttttgat ctcgtgactt
     7141 ttagaaaaac acagggactc gtgggagcaa gcgaggctcc caggaccccc accctctggg
     7201 acaggccctc ccccatgttc ttctgtctcc aggaagggca gcggccctcc catcactgga
     7261 agtctgcagt gggggtcgct gggggtggag agaacactaa gaggtgaaca tgtatgagtg
     7321 tgtgcacgcg tgtgagtgtg catgtgtgtg tgtgcaaagg tccagccacc ccgtcctcca
     7381 gcctgcaagg ggtgtctggc gccttgcctg acacccagcc ccctctcccc ctgagccatt
     7441 gtgggggtcg atcatgaatg tccgaagagt ggccttttcc cgtagccctg cgcccccttt
     7501 ctgtggctgg atggggagac aggtcagggc cccccaccct ctccagcccc tgcagcaaat
     7561 gactactgca cctggacagc ctcctctttt ctagaagtct atttatattg tcattttata
     7621 acactctagc ccctgccctt attgggggac agatggtccc tgtcctgcgg ggtggccctg
     7681 gcagaaccac tgcctgaaga accaggttcc tgcccggtca gcgcagcccc agcccgccca
     7741 cccctgcctc gagttagttt gagctcgcta gcctcgagtc tagagtcgac ctgcaggcat
     7801 gcaagctgat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     7861 tgagcaataa ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca
     7921 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     7981 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     8041 acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca
     8101 agtaaaacct ctacaaatgt ggtaaaatcg aattttaac
//
