LOCUS       sequence-7189-ex        4570 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 4570)
  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 4570)
  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: pAG228 TNKS2 3'UTR mut
FEATURES             Location/Qualifiers
     source          1..4570
                     /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    2525..2646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2536..2555)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2590..2609
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2779..2883
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2884..2952,2953..3744)
                     /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(3102..3121)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3915..4503
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4404..4423
                     /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 ctataccggt gcaaaaggat aaaaatgtga acgaagttta
     1681 acattctgac ttgataaagc tttaataatg tacagtgttt tctaaatatt tcctgttttt
     1741 tcagcacttt aacagatgcc tgaccaggtt aaactgggtt gtctgtacta aattataaac
     1801 agagttaact tgaacctttt atatgttatg cattgattct aacaaactgt aatgccctca
     1861 acagaactaa ttttactaat acaatactgt gttctttaaa acacagcatt tacactgaat
     1921 acaatttcat ttgtaaaact gtaaataaga gcttttgtac tagcccagta tttatttaca
     1981 ttgctttgta atataaatct gttttagaac tgcagcggtt tacaaaattt tttcatatgt
     2041 attgttcatc tatacttcat cttacatcgt catgattgag tgatctttac atttgattcc
     2101 agaggctatg ttcagttgtt agttgggaaa gattgagtta tcagatttaa tttgccgatg
     2161 ggagccttta tctgtcatta gaaatctttc tcatttaaga acttatgaat atgctgaaga
     2221 tttaatttgt gatacctttg tatgtatgag acactgacca aagagctcta actatgatag
     2281 gtcctgatta ctaaagaagc ttctttactg gcctcaattt ctagctttca tgttggaaaa
     2341 ttttctgcag tccttctgtg aaaattagag caaagtgctc ctgtttttta gagaaactaa
     2401 atcttgctgt tgaacaatta ttgtgttctt ttcatggaac ataagtagga tgttaacatt
     2461 tccagggtgg gaagggtaat cctaaatact agtgttctag agcggccgct tcgagcagac
     2521 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     2581 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     2641 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     2701 gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tccaggtggc
     2761 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2821 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2881 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2941 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3001 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3061 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3121 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3181 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3241 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3301 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3361 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3421 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3481 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3541 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3601 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3661 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3721 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3781 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3841 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3901 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3961 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4021 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     4081 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4141 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4201 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4261 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4321 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4381 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4441 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4501 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4561 atggctcgac
//
