LOCUS       sequence-235254-        4232 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 4232)
  AUTHORS   Lim LP, Lau NC, Garrett-Engele P, Grimson A, Schelter JM, Castle J, 
            Bartel DP, Linsley PS, Johnson JM
  TITLE     Microarray analysis shows that some microRNAs downregulate large 
            numbers of target mRNAs.
  JOURNAL   Nature. 2005 Feb 17. 433(7027):769-73.
  PUBMED    15685193
REFERENCE   2  (bases 1 to 4232)
  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: "Nature. 
            2005 Feb 17. 433(7027):769-73."
COMMENT     Sequence Label: pAG34 RELA 3'UTR mut
FEATURES             Location/Qualifiers
     source          1..4232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        138..495
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        138..329
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(161..181)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      346..481
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          565..697
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     680..699
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        742..760
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             770..1705
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2263..2384
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2274..2293)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2328..2347
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2517..2621
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2622..2690,2691..3482)
                     /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(2840..2859)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3653..4232,1..9)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4142..4161
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
       61 gctcacatgg ctcgacagat ctgcgcagca ccatggcctg aaataacctc tgaaagagga
      121 acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
      181 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
      241 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
      301 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
      361 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
      421 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
      481 ctaggctttt gcaaaaagct tgattcttct gacacaacag tctcgaactt aagctgcaga
      541 agttggtcgt gaggcactgg gcaggtaagt atcaaggtta caagacaggt ttaaggagac
      601 caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata ggcacctatt
      661 ggtcttactg acatccactt tgcctttctc tccacaggtg tccactccca gttcaattac
      721 agctcttaag gctagagtac ttaatacgac tcactatagg ctagccacca tgacttcgaa
      781 agtttatgat ccagaacaaa ggaaacggat gataactggt ccgcagtggt gggccagatg
      841 taaacaaatg aatgttcttg attcatttat taattattat gattcagaaa aacatgcaga
      901 aaatgctgtt atttttttac atggtaacgc ggcctcttct tatttatggc gacatgttgt
      961 gccacatatt gagccagtag cgcggtgtat tataccagac cttattggta tgggcaaatc
     1021 aggcaaatct ggtaatggtt cttataggtt acttgatcat tacaaatatc ttactgcatg
     1081 gtttgaactt cttaatttac caaagaagat catttttgtc ggccatgatt ggggtgcttg
     1141 tttggcattt cattatagct atgagcatca agataagatc aaagcaatag ttcacgctga
     1201 aagtgtagta gatgtgattg aatcatggga tgaatggcct gatattgaag aagatattgc
     1261 gttgatcaaa tctgaagaag gagaaaaaat ggttttggag aataacttct tcgtggaaac
     1321 catgttgcca tcaaaaatca tgagaaagtt agaaccagaa gaatttgcag catatcttga
     1381 accattcaaa gagaaaggtg aagttcgtcg tccaacatta tcatggcctc gtgaaatccc
     1441 gttagtaaaa ggtggtaaac ctgacgttgt acaaattgtt aggaattata atgcttatct
     1501 acgtgcaagt gatgatttac caaaaatgtt tattgaatcg gacccaggat tcttttccaa
     1561 tgctattgtt gaaggtgcca agaagtttcc taatactgaa tttgtcaaag taaaaggtct
     1621 tcatttttcg caagaagatg cacctgatga aatgggaaaa tatatcaaat cgttcgttga
     1681 gcgagttctc aaaaatgaac aataattcta ggagctctct ctctcttttt ggaggtgctt
     1741 aagcagaagc attaacttct ctggaaaggg gggagctggg gaaactcaaa cttttcccct
     1801 gtcctgatgg tcagctccct tctctgtagg gaactctggg gtcccccatc cccatcctcc
     1861 agcttctggt actctcctag agacagaagc aggctggagg taaggccttt gagcccacaa
     1921 agccttatca agtgtcttcc atcatggatt cattacagct taatcaaaat aacgccccag
     1981 ataccagccc ctgtatggca ctggcattgt ccctgtcggt aacaccagcg tttgaggggc
     2041 tggccttcct gccctacaga ggtctctgcc ggctctttcc ttgctcaacc atggctgaag
     2101 gaaaccagtg caacagcact ggctctctcc aggatccaga aggggtttgg tctgggactt
     2161 ccttgctctc cctcttctca agtcggttaa tagtagggta agttgttaag agtgggggag
     2221 agcaactagt tgttctagag cggccgcttc gagcagacat gataagatac attgatgagt
     2281 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     2341 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     2401 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     2461 tctacaaatg tggtaaaatc gataaggatc caggtggcac ttttcgggga aatgtgcgcg
     2521 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2581 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2641 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2701 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2761 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2821 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2881 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2941 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3001 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3061 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3121 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     3181 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3241 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3301 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3361 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3421 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3481 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3541 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     3601 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3661 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3721 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3781 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3841 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3901 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3961 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4021 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4081 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4141 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4201 gatttttgtg atgctcgtca ggggggcgga gc
//
