LOCUS       sequence-182928-        7029 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For dual luciferase report assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7029)
  AUTHORS   Hsu AY, Wang D, Gurol T, Zhou W, Zhu X, Lu HY, Deng Q
  TITLE     Overexpression of microRNA-722 fine-tunes neutrophilic inflammation 
            through inhibiting Rac2 in zebrafish.
  JOURNAL   Dis Model Mech. 2017 Sep 27. pii: dmm.030791. doi: 
            10.1242/dmm.030791.
  PUBMED    28954734
REFERENCE   2  (bases 1 to 7029)
  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: "Dis Model 
            Mech. 2017 Sep 27. pii: dmm.030791. doi: 10.1242/dmm.030791."
COMMENT     Sequence Label: pSi-check2-hRac2 3'UTR
FEATURES             Location/Qualifiers
     source          1..7029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..600
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     501..520
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        729..1086
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        729..920
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(752..772)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      937..1072
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          1156..1288
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1271..1290
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1333..1351
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1355..1364
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1361..2296
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    3111..3159
                     /note="synthetic polyadenylation signal"
     promoter        3168..3919
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      3949..3958
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3955..5607
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /db_xref="GI:42718124"
                     /protein_id="AAS38485.1"
                     /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     RBS             4914..4922
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    5651..5772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5662..5681)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5716..5735
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5905..6009
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6010..6078,6079..6870)
                     /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(6228..6247)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
       61 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
      121 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
      181 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      241 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      301 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      361 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      421 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      481 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      541 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
      601 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      661 gctcgacaga tctgcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta
      721 ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
      781 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
      841 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
      901 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
      961 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     1021 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     1081 tgcaaaaagc ttgattcttc tgacacaaca gtctcgaact taagctgcag aagttggtcg
     1141 tgaggcactg ggcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa
     1201 ctgggcttgt cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact
     1261 gacatccact ttgcctttct ctccacaggt gtccactccc agttcaatta cagctcttaa
     1321 ggctagagta cttaatacga ctcactatag gctagccacc atggcttcca aggtgtacga
     1381 ccccgagcaa cgcaaacgca tgatcactgg gcctcagtgg tgggctcgct gcaagcaaat
     1441 gaacgtgctg gactccttca tcaactacta tgattccgag aagcacgccg agaacgccgt
     1501 gatttttctg catggtaacg ctgcctccag ctacctgtgg aggcacgtcg tgcctcacat
     1561 cgagcccgtg gctagatgca tcatccctga tctgatcgga atgggtaagt ccggcaagag
     1621 cgggaatggc tcatatcgcc tcctggatca ctacaagtac ctcaccgctt ggttcgagct
     1681 gctgaacctt ccaaagaaaa tcatctttgt gggccacgac tggggggctt gtctggcctt
     1741 tcactactcc tacgagcacc aagacaagat caaggccatc gtccatgctg agagtgtcgt
     1801 ggacgtgatc gagtcctggg acgagtggcc tgacatcgag gaggatatcg ccctgatcaa
     1861 gagcgaagag ggcgagaaaa tggtgcttga gaataacttc ttcgtcgaga ccatgctccc
     1921 aagcaagatc atgcggaaac tggagcctga ggagttcgct gcctacctgg agccattcaa
     1981 ggagaagggc gaggttagac ggcctaccct ctcctggcct cgcgagatcc ctctcgttaa
     2041 gggaggcaag cccgacgtcg tccagattgt ccgcaactac aacgcctacc ttcgggccag
     2101 cgacgatctg cctaagatgt tcatcgagtc cgaccctggg ttcttttcca acgctattgt
     2161 cgagggagct aagaagttcc ctaacaccga gttcgtgaag gtgaagggcc tccacttcag
     2221 ccaggaggac gctccagatg aaatgggtaa gtacatcaag agcttcgtgg agcgcgtgct
     2281 gaagaacgag cagtaattct aggcgatcgc tcgagggttg caccccagcg ctcccaccta
     2341 gatgggtctg atcctccagg atccccaccc aaagcctgat ggcaccccgg ctggccatgc
     2401 tgtcccctcc ctgtggcgtt tcttagcaga tggctgcaga gctttgttga tggtcttttc
     2461 tgtactggag gcctcctgag gccaggaacg tgcaaatttg caggtgctgc atcccaagcc
     2521 cctcatgctc ctgccttcct gagggccaga ggggagcccc aggacccatt aagccacccc
     2581 cgtgttcctg ccgtcagtgc caactgccgc atgtggaagc atctacccgt tcactccagt
     2641 cccaccccac gcctgactcc cctctggaaa ctgcaggcca gatggttgct gccacaactt
     2701 gtgtaccttc agggatgggg ctcttactcc ctcctgaggc cagctgctct aatatcgatg
     2761 gtcctgcttg ccagagagtt cctctaccca gcaaaaatga gtgtctcaga agtgtgctcc
     2821 tctggcctca gttctcctct tttggaacaa cataaaacaa atttaatttt ctacgcctct
     2881 ggggatatct gctcagccaa tggaaaatct gggttcaacc agcccctgcc atttcttaag
     2941 actttctgct gcactcacag gatcctgagc tgcacttacc tgtgagagtc ttcaaacttt
     3001 taaaccttgc cagtcaggac ttttgctatt gcaaatagaa aacccaactc aacctgctta
     3061 agcagaaaat aaatttattg attcaagttt ggagagcggc cgctggccgc aataaaatat
     3121 ctttattttc attacatctg tgtgttggtt ttttgtgtga ggatctaaat gagtcttcgg
     3181 acctcgcggg ggccgcttaa gcggtggtta gggtttgtct gacgcggggg gagggggaag
     3241 gaacgaaaca ctctcattcg gaggcggctc ggggtttggt cttggtggcc acgggcacgc
     3301 agaagagcgc cgcgatcctc ttaagcaccc ccccgccctc cgtggaggcg ggggtttggt
     3361 cggcgggtgg taactggcgg gccgctgact cgggcgggtc gcgcgcccca gagtgtgacc
     3421 ttttcggtct gctcgcagac ccccgggcgg cgccgccgcg gcggcgacgg gctcgctggg
     3481 tcctaggctc catggggacc gtatacgtgg acaggctctg gagcatccgc acgactgcgg
     3541 tgatattacc ggagaccttc tgcgggacga gccgggtcac gcggctgacg cggagcgtcc
     3601 gttgggcgac aaacaccagg acggggcaca ggtacactat cttgtcaccc ggaggcgcga
     3661 gggactgcag gagcttcagg gagtggcgca gctgcttcat ccccgtggcc cgttgctcgc
     3721 gtttgctggc ggtgtccccg gaagaaatat atttgcatgt ctttagttct atgatgacac
     3781 aaaccccgcc cagcgtcttg tcattggcga attcgaacac gcagatgcag tcggggcggc
     3841 gcggtcccag gtccacttcg catattaagg tgacgcgtgt ggcctcgaac accgagcgac
     3901 cctgcagcga cccgcttaaa agcttggcat tccggtactg ttggtaaagc caccatggcc
     3961 gatgctaaga acattaagaa gggccctgct cccttctacc ctctggagga tggcaccgct
     4021 ggcgagcagc tgcacaaggc catgaagagg tatgccctgg tgcctggcac cattgccttc
     4081 accgatgccc acattgaggt ggacatcacc tatgccgagt acttcgagat gtctgtgcgc
     4141 ctggccgagg ccatgaagag gtacggcctg aacaccaacc accgcatcgt ggtgtgctct
     4201 gagaactctc tgcagttctt catgccagtg ctgggcgccc tgttcatcgg agtggccgtg
     4261 gcccctgcta acgacattta caacgagcgc gagctgctga acagcatggg catttctcag
     4321 cctaccgtgg tgttcgtgtc taagaagggc ctgcagaaga tcctgaacgt gcagaagaag
     4381 ctgcctatca tccagaagat catcatcatg gactctaaga ccgactacca gggcttccag
     4441 agcatgtaca cattcgtgac atctcatctg cctcctggct tcaacgagta cgacttcgtg
     4501 ccagagtctt tcgacaggga caaaaccatt gccctgatca tgaacagctc tgggtctacc
     4561 ggcctgccta agggcgtggc cctgcctcat cgcaccgcct gtgtgcgctt ctctcacgcc
     4621 cgcgacccta ttttcggcaa ccagatcatc cccgacaccg ctattctgag cgtggtgcca
     4681 ttccaccacg gcttcggcat gttcaccacc ctgggctacc tgatttgcgg ctttcgggtg
     4741 gtgctgatgt accgcttcga ggaggagctg ttcctgcgca gcctgcaaga ctacaaaatt
     4801 cagtctgccc tgctggtgcc aaccctgttc agcttcttcg ctaagagcac cctgatcgac
     4861 aagtacgacc tgtctaacct gcacgagatt gcctctggcg gcgccccact gtctaaggag
     4921 gtgggcgaag ccgtggccaa gcgctttcat ctgccaggca tccgccaggg ctacggcctg
     4981 accgagacaa ccagcgccat tctgattacc ccagagggcg acgacaagcc tggcgccgtg
     5041 ggcaaggtgg tgccattctt cgaggccaag gtggtggacc tggacaccgg caagaccctg
     5101 ggagtgaacc agcgcggcga gctgtgtgtg cgcggcccta tgattatgtc cggctacgtg
     5161 aataaccctg aggccacaaa cgccctgatc gacaaggacg gctggctgca ctctggcgac
     5221 attgcctact gggacgagga cgagcacttc ttcatcgtgg accgcctgaa gtctctgatc
     5281 aagtacaagg gctaccaggt ggccccagcc gagctggagt ctatcctgct gcagcaccct
     5341 aacattttcg acgccggagt ggccggcctg cccgacgacg atgccggcga gctgcctgcc
     5401 gccgtcgtcg tgctggaaca cggcaagacc atgaccgaga aggagatcgt ggactatgtg
     5461 gccagccagg tgacaaccgc caagaagctg cgcggcggag tggtgttcgt ggacgaggtg
     5521 cccaagggcc tgaccggcaa gctggacgcc cgcaagatcc gcgagatcct gatcaaggct
     5581 aagaaaggcg gcaagatcgc cgtgtaataa ttctagagtc ggggcggccg gccgcttcga
     5641 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     5701 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     5761 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     5821 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatcca
     5881 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5941 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6001 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6061 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6121 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6181 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6241 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6301 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6361 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6421 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6481 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6541 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6601 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6661 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6721 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6781 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6841 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6901 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6961 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7021 gaaaagatc
//
