LOCUS       sequence-7200-ex        4137 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 4137)
  AUTHORS   Farh KK, Grimson A, Jan C, Lewis BP, Johnston WK, Lim LP, Burge CB, 
            Bartel DP
  TITLE     The widespread impact of mammalian MicroRNAs on mRNA repression and 
            evolution.
  JOURNAL   Science. 2005 Dec 16. 310(5755):1817-21.
  PUBMED    16308420
REFERENCE   2  (bases 1 to 4137)
  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: "Science. 
            2005 Dec 16. 310(5755):1817-21."
COMMENT     Sequence Label: pAG75 ARHGEF18 3'UTR mut
FEATURES             Location/Qualifiers
     source          1..4137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    452..573
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(463..482)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     517..536
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        706..810
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(811..879,880..1671)
                     /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(1029..1048)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1842..2430
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2331..2350
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2559..2916
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2559..2750
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2582..2602)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2767..2902
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          2986..3118
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3101..3120
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        3163..3181
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3191..4126
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
ORIGIN
        1 ccgctgaata cagtgaacag ggagaatgcc gccactcggg gacagatggg cacaagggag
       61 gggaaactcc atcaggaagt gctcccctgg gcagaggcgc ccactgggtg ctgtgggctc
      121 aggagggggc ggggcaggag ctggtgccaa ccgggaacca gagccccaca gccatacagc
      181 ccattggtga caaggtcctg agaacacagt ggccaggtgt ccccaggctc ctggcccctc
      241 cgacgacctc aactctgccc agcccggtcc ctggccatca gcgacgctgt ccgccccccg
      301 tcagatccca tgtgtgccat gtttatcatc agtgttttgt atttttgtac tgagtatcgg
      361 agcactttac agaagctgac tgtacattcc tgttctgttg tgaagagaag aatgccagac
      421 cctctagagc ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
      481 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
      541 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
      601 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
      661 ggtaaaatcg ataaggatcc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
      721 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
      781 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
      841 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
      901 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
      961 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1021 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     1081 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1141 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     1201 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     1261 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     1321 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     1381 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     1441 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     1501 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     1561 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     1621 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     1681 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     1741 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     1801 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1861 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1921 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1981 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2041 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2101 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2161 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2221 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2281 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2341 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2401 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2461 ctggcctttt gctggccttt tgctcacatg gctcgacaga tctgcgcagc accatggcct
     2521 gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg
     2581 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2641 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     2701 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     2761 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     2821 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     2881 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttgattcttc tgacacaaca
     2941 gtctcgaact taagctgcag aagttggtcg tgaggcactg ggcaggtaag tatcaaggtt
     3001 acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg
     3061 cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt
     3121 gtccactccc agttcaatta cagctcttaa ggctagagta cttaatacga ctcactatag
     3181 gctagccacc atgacttcga aagtttatga tccagaacaa aggaaacgga tgataactgg
     3241 tccgcagtgg tgggccagat gtaaacaaat gaatgttctt gattcattta ttaattatta
     3301 tgattcagaa aaacatgcag aaaatgctgt tattttttta catggtaacg cggcctcttc
     3361 ttatttatgg cgacatgttg tgccacatat tgagccagta gcgcggtgta ttataccaga
     3421 ccttattggt atgggcaaat caggcaaatc tggtaatggt tcttataggt tacttgatca
     3481 ttacaaatat cttactgcat ggtttgaact tcttaattta ccaaagaaga tcatttttgt
     3541 cggccatgat tggggtgctt gtttggcatt tcattatagc tatgagcatc aagataagat
     3601 caaagcaata gttcacgctg aaagtgtagt agatgtgatt gaatcatggg atgaatggcc
     3661 tgatattgaa gaagatattg cgttgatcaa atctgaagaa ggagaaaaaa tggttttgga
     3721 gaataacttc ttcgtggaaa ccatgttgcc atcaaaaatc atgagaaagt tagaaccaga
     3781 agaatttgca gcatatcttg aaccattcaa agagaaaggt gaagttcgtc gtccaacatt
     3841 atcatggcct cgtgaaatcc cgttagtaaa aggtggtaaa cctgacgttg tacaaattgt
     3901 taggaattat aatgcttatc tacgtgcaag tgatgattta ccaaaaatgt ttattgaatc
     3961 ggacccagga ttcttttcca atgctattgt tgaaggtgcc aagaagtttc ctaatactga
     4021 atttgtcaaa gtaaaaggtc ttcatttttc gcaagaagat gcacctgatg aaatgggaaa
     4081 atatatcaaa tcgttcgttg agcgagttct caaaaatgaa caataattct aggagct
//
