LOCUS       sequence-7188-ex        4109 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 4109)
  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 4109)
  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: pAG227 CORO1C 3'UTR mut
FEATURES             Location/Qualifiers
     source          1..4109
                     /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    2064..2185
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2075..2094)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2129..2148
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2318..2422
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2423..2491,2492..3283)
                     /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(2641..2660)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3454..4042
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3943..3962
                     /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 cgagctggtt attggtgtgg tcctagggag ggcggaaagg
     1681 gaggcactgc catttggaga ctgaccattt cagatttgtc aaccagcgat aggccactga
     1741 ccagtaagaa ctcaatttgt ctcccaaatt tgcagaaaca aaacgtgatt taaaagctga
     1801 gctttttatc agaaagcttt tttgatgttt taagtgttat gtgacttgtt gaacttttta
     1861 aaaagtgcta cttttaaaat cccagatact ctgaatttta gaaaacaaac taattctgat
     1921 tgtgtcgtgc ccaagtaccc ttttttttta atgaataggg accaatgcca cattgctttt
     1981 tatatttctt tcttttttaa tgttgccaaa accaaaagta gctttgacta gtgttctaga
     2041 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     2101 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     2161 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     2221 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat
     2281 cgataaggat ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2341 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2401 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2461 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2521 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2581 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     2641 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     2701 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     2761 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     2821 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     2881 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     2941 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3001 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3061 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3121 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3181 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3241 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3301 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3361 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3421 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3481 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3541 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3601 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3661 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3721 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3781 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3841 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3901 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3961 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4021 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4081 ttgctggcct tttgctcaca tggctcgac
//
