LOCUS       sequence-7204-ex        3918 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 3918)
  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 3918)
  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: pAG85 3'UTR mut
FEATURES             Location/Qualifiers
     source          1..3918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    233..354
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(244..263)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     298..317
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        487..591
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(592..660,661..1452)
                     /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(810..829)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1623..2211
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2112..2131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2340..2697
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2340..2531
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2363..2383)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2548..2683
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          2767..2899
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2882..2901
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        2944..2962
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2972..3907
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
ORIGIN
        1 cagtttccta cgtgtgcgag aatgctaagg tcttcaagtt tcctattctg ttttgtgctg
       61 tgtgaaagtt ctgatatgac ccctgatgat aaagtccata actgtagtta actggtcatc
      121 actgctcagg tcccagttac taagaagact accaaaatgg catcatctta atcttgaagc
      181 gtatttctga gaatgctgaa ccgtctagag cggccgcttc gagcagacat gataagatac
      241 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
      301 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
      361 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
      421 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc caggtggcac ttttcgggga
      481 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
      541 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
      601 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
      661 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
      721 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
      781 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
      841 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
      901 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
      961 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     1021 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     1081 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     1141 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     1201 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     1261 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     1321 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     1381 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     1441 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     1501 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     1561 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     1621 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     1681 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     1741 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     1801 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     1861 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     1921 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     1981 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     2041 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     2101 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     2161 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     2221 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat ggctcgacag
     2281 atctgcgcag caccatggcc tgaaataacc tctgaaagag gaacttggtt aggtaccttc
     2341 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     2401 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     2461 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2521 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     2581 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     2641 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     2701 cttgattctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc gtgaggcact
     2761 gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg
     2821 tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac tgacatccac
     2881 tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta aggctagagt
     2941 acttaatacg actcactata ggctagccac catgacttcg aaagtttatg atccagaaca
     3001 aaggaaacgg atgataactg gtccgcagtg gtgggccaga tgtaaacaaa tgaatgttct
     3061 tgattcattt attaattatt atgattcaga aaaacatgca gaaaatgctg ttattttttt
     3121 acatggtaac gcggcctctt cttatttatg gcgacatgtt gtgccacata ttgagccagt
     3181 agcgcggtgt attataccag accttattgg tatgggcaaa tcaggcaaat ctggtaatgg
     3241 ttcttatagg ttacttgatc attacaaata tcttactgca tggtttgaac ttcttaattt
     3301 accaaagaag atcatttttg tcggccatga ttggggtgct tgtttggcat ttcattatag
     3361 ctatgagcat caagataaga tcaaagcaat agttcacgct gaaagtgtag tagatgtgat
     3421 tgaatcatgg gatgaatggc ctgatattga agaagatatt gcgttgatca aatctgaaga
     3481 aggagaaaaa atggttttgg agaataactt cttcgtggaa accatgttgc catcaaaaat
     3541 catgagaaag ttagaaccag aagaatttgc agcatatctt gaaccattca aagagaaagg
     3601 tgaagttcgt cgtccaacat tatcatggcc tcgtgaaatc ccgttagtaa aaggtggtaa
     3661 acctgacgtt gtacaaattg ttaggaatta taatgcttat ctacgtgcaa gtgatgattt
     3721 accaaaaatg tttattgaat cggacccagg attcttttcc aatgctattg ttgaaggtgc
     3781 caagaagttt cctaatactg aatttgtcaa agtaaaaggt cttcattttt cgcaagaaga
     3841 tgcacctgat gaaatgggaa aatatatcaa atcgttcgtt gagcgagttc tcaaaaatga
     3901 acaataattc taggagct
//
