LOCUS       sequence-7193-ex        4408 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 4408)
  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 4408)
  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: pAG177 HIPK1 3'UTR mut
FEATURES             Location/Qualifiers
     source          1..4408
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        62..253
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(85..105)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      270..405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          489..621
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     604..623
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        666..684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             694..1629
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2363..2484
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2374..2393)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2428..2447
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2617..2721
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2722..2790,2791..3582)
                     /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(2940..2959)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3753..4341
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4242..4261
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
       61 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      421 agcttgattc ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca
      481 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      541 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      601 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
      661 gtacttaata cgactcacta taggctagcc accatgactt cgaaagttta tgatccagaa
      721 caaaggaaac ggatgataac tggtccgcag tggtgggcca gatgtaaaca aatgaatgtt
      781 cttgattcat ttattaatta ttatgattca gaaaaacatg cagaaaatgc tgttattttt
      841 ttacatggta acgcggcctc ttcttattta tggcgacatg ttgtgccaca tattgagcca
      901 gtagcgcggt gtattatacc agaccttatt ggtatgggca aatcaggcaa atctggtaat
      961 ggttcttata ggttacttga tcattacaaa tatcttactg catggtttga acttcttaat
     1021 ttaccaaaga agatcatttt tgtcggccat gattggggtg cttgtttggc atttcattat
     1081 agctatgagc atcaagataa gatcaaagca atagttcacg ctgaaagtgt agtagatgtg
     1141 attgaatcat gggatgaatg gcctgatatt gaagaagata ttgcgttgat caaatctgaa
     1201 gaaggagaaa aaatggtttt ggagaataac ttcttcgtgg aaaccatgtt gccatcaaaa
     1261 atcatgagaa agttagaacc agaagaattt gcagcatatc ttgaaccatt caaagagaaa
     1321 ggtgaagttc gtcgtccaac attatcatgg cctcgtgaaa tcccgttagt aaaaggtggt
     1381 aaacctgacg ttgtacaaat tgttaggaat tataatgctt atctacgtgc aagtgatgat
     1441 ttaccaaaaa tgtttattga atcggaccca ggattctttt ccaatgctat tgttgaaggt
     1501 gccaagaagt ttcctaatac tgaatttgtc aaagtaaaag gtcttcattt ttcgcaagaa
     1561 gatgcacctg atgaaatggg aaaatatatc aaatcgttcg ttgagcgagt tctcaaaaat
     1621 gaacaataat tctaggagct ctttccattt tggattctcg gctgagttct cacagaagca
     1681 ttttccccat gtggctctct cactgtgcgt tgctaccttg cttctgtgag aattcaggaa
     1741 gcaggtgaga ggagtcaagc caatattaaa tatgcattct tttaaagtat gtgcaatcac
     1801 ttttagaatg aatttttttt tccttttccc atgtggcagt ccttcctgca catagttgac
     1861 tgacctagta aaatatttgc ttgttgaaaa aaacatgtta acagatgtgt ttataccaaa
     1921 gagcctgttg tattgcttac catgtcccca tactatgagg agaagttttg tggtgccgct
     1981 ggtgacaagg aactcacaga aaggtttctt agctggtgaa gaatatagag aaggaaccaa
     2041 agcctgttga gtcattgagg cttttgaggt ttctttttta acagcttgta tagtcttggg
     2101 gcccttcaag ctgtgaaatt gtccttgtac tctcagctcc tgcatggatc tgggtcaagt
     2161 agaaggtact ggggatgggg actgacctgc ccataaagga tttggggaaa gaagattaat
     2221 cctaaaatac aggtgtgttc catctgaatt gaaaatgata tatttgagat ataattttag
     2281 gactggttct gtgtagatag agatggtgtc aaggaggtgc aggatactag tgttctagag
     2341 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     2401 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     2461 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     2521 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
     2581 gataaggatc caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2641 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2701 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     2761 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     2821 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     2881 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     2941 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     3001 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3061 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3121 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3181 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3241 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     3301 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3361 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     3421 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     3481 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     3541 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     3601 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     3661 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3721 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3781 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3841 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     3901 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3961 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4021 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4081 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4141 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4201 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4261 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4321 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4381 tgctggcctt ttgctcacat ggctcgac
//
