LOCUS       sequence-7190-ex        4378 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 4378)
  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 4378)
  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: pAG174 TNS3 3'UTR mut
FEATURES             Location/Qualifiers
     source          1..4378
                     /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    2333..2454
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2344..2363)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2398..2417
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2587..2691
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2692..2760,2761..3552)
                     /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(2910..2929)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3723..4311
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4212..4231
                     /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 catgtgtgag cagaagggag gatgaggaaa aagagaagaa
     1681 accccggtac tgacaagctg tttttgagtg ccactgtttg ccatcatcta agccactgaa
     1741 tcaagtgtat ttcaggctta tttcaactga ccaatgccct ggttttcctg cttgaatctg
     1801 ttcgtggtca aaggtttggg ggaatttgtg accctggaac atccccagag tgaaagatgg
     1861 agctgggcca catcagaata aggccttggc cccatcctct cacagcctag gtgctctgca
     1921 ggcatgctga ctgtcctgat tgcgatccag cccgaaattc cctcctctgc tttcaaaagt
     1981 caaatccccc attcttaggc cacactggtg tcacaagctc ctgtcaggga gctggggttt
     2041 gggaatgtgc tttgtgaact ctgctttaaa gtgaggggcc gaggaaaact tagaaacagg
     2101 cagagttgga agcagccaaa tcacagtggg tgttgtgtgt gtgtgcgtgt gtgcatgcgt
     2161 gcgtgtatgc gtgtgtgaaa gcaggtggac ctgaccactt tttagctcct attgatgcac
     2221 caaaccaagt gcctcatttc tgtgccaaat gtttgccttg gtcgttgtgg acctccttct
     2281 ctaacttgcg gtggcactag tgttctagag cggccgcttc gagcagacat gataagatac
     2341 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2401 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2461 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     2521 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc caggtggcac ttttcgggga
     2581 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2641 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     2701 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2761 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2821 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2881 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2941 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3001 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3061 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3121 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3181 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3241 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3301 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3361 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3421 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3481 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3541 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3601 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3661 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3721 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3781 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3841 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     3901 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3961 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4021 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4081 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4141 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4201 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4261 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4321 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat ggctcgac
//
