LOCUS       sequence-214936-        6759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3'UTR reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6759)
  AUTHORS   Li X, Zhao F, Xin M, Li G, Luna C, Li G, Zhou Q, He Y, Yu B, Olson 
            E, Gonzalez P, Wang S
  TITLE     Regulation of intraocular pressure by microRNA cluster miR-143/145.
  JOURNAL   Sci Rep. 2017 Apr 19;7(1):915. doi: 10.1038/s41598-017-01003-z.
  PUBMED    28424493
REFERENCE   2  (bases 1 to 6759)
  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: "Sci Rep."; 
            date: "2017-04-19"; volume: "7(1):915. doi"; pages: " 
            10.1038/s41598-017-01003-z"
COMMENT     Sequence Label: pmiR-report-hARPC2
FEATURES             Location/Qualifiers
     source          1..6759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        60..363
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        364..567
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     518..538
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             592..2241
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIVPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(658..676)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(2686..2703)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2686..2702)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2695..2717)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2911..2930)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3092..3408
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3259..3394
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3505..4104
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3505..3524)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4001..4021
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4334..4415
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4371..4390)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4503..4607
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4608..4676,4677..5468)
                     /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(4826..4845)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5639..6227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6128..6147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6381..6398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6515..6536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6551..6556,6557..6574,6575..6581)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6589..6605
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6594..6616
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6613..6629
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6613..6629
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6649..6667
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6649..6666
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     6679..6698
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6679..6697
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
       61 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      121 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      181 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      241 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      301 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      361 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      421 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      481 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      541 cggtgggagg tctatataag cagagctggt ttagtgaacc gtggatccac catggaagac
      601 gccaaaaaca taaagaaagg cccggcgcca ttctatcctc tagaggatgg aaccgctgga
      661 gagcaactgc ataaggctat gaagagatac gccctggttc ctggaacaat tgcttttaca
      721 gatgcacata tcgaggtgaa catcacgtac gcggaatact tcgaaatgtc cgttcggttg
      781 gcagaagcta tgaaacgata tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa
      841 aactctcttc aattctttat gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg
      901 cccgcgaacg acatttataa tgaacgtgaa ttgctcaaca gtatgaacat ttcgcagcct
      961 accgtagtgt ttgtttccaa aaaggggttg caaaaaattt tgaacgtgca aaaaaaatta
     1021 ccaataatcc agaaaattat tatcatggat tctaaaacgg attaccaggg atttcagtcg
     1081 atgtacacgt tcgtcacatc tcatctacct cccggtttta atgaatacga ttttgtacca
     1141 gagtcctttg atcgtgacaa aacaattgca ctgataatga attcctctgg atctactggg
     1201 ttacctaagg gtgtggccct tccgcataga actgcctgcg tcagattctc gcatgccaga
     1261 gatcctattt ttggcaatca aatcgttccg gatactgcga ttttaagtgt tgtcccattc
     1321 catcacggtt ttggaatgtt tactacactc ggatatttga tatgtggatt tcgagtcgtc
     1381 ttaatgtata gatttgaaga agagctgttt ttacgatccc ttcaggatta caaaattcaa
     1441 agtgcgttgc tagtaccaac cctattttca ttcttcgcca aaagcactct gattgacaaa
     1501 tacgatttat ctaatttaca cgaaattgct tctgggggcg cacctctttc gaaagaagtc
     1561 ggggaagcgg ttgcaaaacg cttccatctt ccagggatac gacaaggata tgggctcact
     1621 gagactacat cagctattct gattacaccc gagggggatg ataaaccggg cgcggtcggt
     1681 aaagttgttc cattttttga agcgaaggtt gtggatctgg ataccgggaa aacgctgggc
     1741 gttaatcaga gaggcgaatt atgtgtcaga ggacctatga ttatgtccgg ttatgtaaac
     1801 aatccggaag cgaccaacgc cttgattgac aaggatggat ggctacattc tggagacata
     1861 gcttactggg acgaagacga acacttcttc atagttgacc gcttgaagtc tttaattaaa
     1921 tacaaaggat atcaggtggc ccccgctgaa ttggaatcga tattgttaca acaccccaac
     1981 atcttcgacg cgggcgtggc aggtcttccc gacgatgacg ccggtgaact tcccgccgcc
     2041 gttgttgttt tggagcacgg aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc
     2101 agtcaagtaa caaccgcgaa aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg
     2161 aaaggtctta ccggaaaact cgacgcaaga aaaatcagag agatcctcat aaaggccaag
     2221 aagggcggaa agtccaaatt gctcgagtga tgaaagctgc gcactagtga gggagctcct
     2281 tgggaataag aggaggaagc ggctggcaac tgaaggctgg aacacttgct actggataat
     2341 cgtagctttt aatgttgcgc ctcttcaggt tcttaaggga ttctccgttt tggttccatt
     2401 ttgtacacgt ttggaaaata atctgcagaa acgagctgtg cttgcaaaga cttcatagtt
     2461 cccaagaatt aaaaaaaaaa aaaaaagaat tccacttgat caacttaatt ccttttcttt
     2521 atcttccctc cctcacttcc cttttctccc accctctttt ccaagctgtt tcgctttgca
     2581 atatattact ggtaatgagt tgcaggataa tgcaagctta ataaaggatc ttttattttc
     2641 attggatctg tgtgttggtt ttttgtatgc ggccgctagc ttggcactgg ccgtcgtttt
     2701 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     2761 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2821 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2881 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     2941 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3001 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3061 gtcatcaccg aaacgcgcga gacgaaagat tggtgtggaa agtccccagg ctccccagca
     3121 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3181 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3241 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3301 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3361 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctagcttgc
     3421 atgcctgcag gtcggccgcc acgaccggtg ccgccaccat cccctgaccc acgcccctga
     3481 cccctcacaa ggagacgacc ttccatgacc gagtacaagc ccacggtgcg cctcgccacc
     3541 cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     3601 acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     3661 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     3721 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     3781 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     3841 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     3901 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     3961 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     4021 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     4081 atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg
     4141 agcgcacgac cccatggctc cgaccgaagc cacccggggc ggccccgccg accccgcacc
     4201 cgcccccgag gcccaccgac tctagaggat cataatcagc cataccacat ttgtagaggt
     4261 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     4321 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4381 cacaaatttc acaaataaag catttttttc actgcaatct cgtgatacgc ctatttttat
     4441 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     4501 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4561 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4621 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4681 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4741 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4801 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4861 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4921 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4981 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5041 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5101 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5161 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5221 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5281 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5341 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5401 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5461 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5521 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5581 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5641 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5701 cggtggtttt tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5761 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5821 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5881 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5941 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6001 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6061 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6301 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     6361 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     6421 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     6481 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     6541 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     6601 aacaatttca cacaggaaac agctatgaca tgattacgaa ttgcaacgat ttaggtgaca
     6661 ctatagaaga gaaggaatta atacgactca ctatagggag agagagagaa ttaccctcac
     6721 taaagggagg agaagcatga attcaaggta ccagatctt
//
