LOCUS       sequence-7174-ex        4710 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 4710)
  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 4710)
  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: pAG186 NID1 3'UTR
FEATURES             Location/Qualifiers
     source          1..4710
                     /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    2665..2786
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2676..2695)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2730..2749
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2919..3023
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3024..3092,3093..3884)
                     /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(3242..3261)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4055..4643
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4544..4563
                     /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 cgttgccctg acaacacctt gggagttgac
     1681 tgtatcgaac ggaaatgaag acaagagtgc cttatttcct ttccaagtat ttcacagcaa
     1741 cactctactt gaagcaactt ggtccagatt gaaaagtgtc ctctggctga gtggccacta
     1801 ggcccagacc cagcccagcc tgagccccaa caacaacttt tccctcactg ttccccaaaa
     1861 catgcaccct ggacttctct aatagaaaag tctccacccc tacacaagga cagaaccctc
     1921 cacccctacc cccaaccctc agacagactt atacacccct gagtgaggat tacatgccca
     1981 tcccagtgtc ctaggacctt ttcccaatac tagcccccca gtggtgaaca gaacctccca
     2041 aatttgagtt gcacccttcc ctgtggcctt atgagctcag cctcgctttg aggtacccac
     2101 cgtcctgtca gctccttgac ctatgagctg gggcctgact aggaaaagtt gggagttaag
     2161 gaggaaatta gcattcctta atgttttgtt ttggtgctct gaatttcttc tttattatag
     2221 tcctatagtt ttactcctca gttcctcacc atcatcatct tgtctaagac ccccattata
     2281 atattcatgc gctgcttttt catcaaaacc taccctgtcc tagagatcta tgggcatttg
     2341 gtggatgata atgagcagcc cctcccagat agaatgtcaa tatttgagca gtaggatatt
     2401 ggcatttgtt agttaaaggc ttaaatcaaa agaatgtcca atggtaggaa tttcaaggtg
     2461 taggtcagat atttgagaat aggggatttt tttgatgtgc cttaaattat accaaagatt
     2521 actaattatt cctctttgcc caaaatactt gcatccaagg ttctagtctc tgttgctgtg
     2581 ctggtcttta gccccactgc tggcactgat gtccctcctt tttcacgact agtgttctag
     2641 agcggccgct tcgagcagac atgataagat acattgatga gtttggacaa accacaacta
     2701 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     2761 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     2821 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa
     2881 tcgataagga tccaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2941 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3001 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3061 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3121 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3181 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3241 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3301 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3361 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3421 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3481 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3541 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     3601 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3661 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3721 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3781 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3841 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     3901 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     3961 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4021 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4081 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4141 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4201 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4261 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4321 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4381 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4441 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4501 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4561 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4621 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4681 tttgctggcc ttttgctcac atggctcgac
//
