LOCUS       sequence-235253-        4262 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 4262)
  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 4262)
  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: pIS50 KIF2 3'UTR
FEATURES             Location/Qualifiers
     source          1..4262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        664..1415
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1485..1617
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1600..1619
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1662..1680
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1690..2625
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2873..2994
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2884..2903)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2938..2957
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3127..3231
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3232..3300,3301..4092)
                     /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(3450..3469)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg ctcgacagat
      661 ctaaatgagt cttcggacct cgcgggggcc gcttaagcgg tggttagggt ttgtctgacg
      721 cggggggagg gggaaggaac gaaacactct cattcggagg cggctcgggg tttggtcttg
      781 gtggccacgg gcacgcagaa gagcgccgcg atcctcttaa gcaccccccc gccctccgtg
      841 gaggcggggg tttggtcggc gggtggtaac tggcgggccg ctgactcggg cgggtcgcgc
      901 gccccagagt gtgacctttt cggtctgctc gcagaccccc gggcggcgcc gccgcggcgg
      961 cgacgggctc gctgggtcct aggctccatg gggaccgtat acgtggacag gctctggagc
     1021 atccgcacga ctgcggtgat attaccggag accttctgcg ggacgagccg ggtcacgcgg
     1081 ctgacgcgga gcgtccgttg ggcgacaaac accaggacgg ggcacaggta cactatcttg
     1141 tcacccggag gcgcgaggga ctgcaggagc ttcagggagt ggcgcagctg cttcatcccc
     1201 gtggcccgtt gctcgcgttt gctggcggtg tccccggaag aaatatattt gcatgtcttt
     1261 agttctatga tgacacaaac cccgcccagc gtcttgtcat tggcgaattc gaacacgcag
     1321 atgcagtcgg ggcggcgcgg tcccaggtcc acttcgcata ttaaggtgac gcgtgtggcc
     1381 tcgaacaccg agcgaccctg cagcgacccg cttaaaagct tgattcttct gacacaacag
     1441 tctcgaactt aagctgcaga agttggtcgt gaggcactgg gcaggtaagt atcaaggtta
     1501 caagacaggt ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc
     1561 gtttctgata ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg
     1621 tccactccca gttcaattac agctcttaag gctagagtac ttaatacgac tcactatagg
     1681 ctagccacca tgacttcgaa agtttatgat ccagaacaaa ggaaacggat gataactggt
     1741 ccgcagtggt gggccagatg taaacaaatg aatgttcttg attcatttat taattattat
     1801 gattcagaaa aacatgcaga aaatgctgtt atttttttac atggtaacgc ggcctcttct
     1861 tatttatggc gacatgttgt gccacatatt gagccagtag cgcggtgtat tataccagac
     1921 cttattggta tgggcaaatc aggcaaatct ggtaatggtt cttataggtt acttgatcat
     1981 tacaaatatc ttactgcatg gtttgaactt cttaatttac caaagaagat catttttgtc
     2041 ggccatgatt ggggtgcttg tttggcattt cattatagct atgagcatca agataagatc
     2101 aaagcaatag ttcacgctga aagtgtagta gatgtgattg aatcatggga tgaatggcct
     2161 gatattgaag aagatattgc gttgatcaaa tctgaagaag gagaaaaaat ggttttggag
     2221 aataacttct tcgtggaaac catgttgcca tcaaaaatca tgagaaagtt agaaccagaa
     2281 gaatttgcag catatcttga accattcaaa gagaaaggtg aagttcgtcg tccaacatta
     2341 tcatggcctc gtgaaatccc gttagtaaaa ggtggtaaac ctgacgttgt acaaattgtt
     2401 aggaattata atgcttatct acgtgcaagt gatgatttac caaaaatgtt tattgaatcg
     2461 gacccaggat tcttttccaa tgctattgtt gaaggtgcca agaagtttcc taatactgaa
     2521 tttgtcaaag taaaaggtct tcatttttcg caagaagatg cacctgatga aatgggaaaa
     2581 tatatcaaat cgttcgttga gcgagttctc aaaaatgaac aataattcta ggagctctga
     2641 ccctaaattt ctgaagcaca gccaagaaaa attacattcc ttgtcattgt aaattacctt
     2701 tgtgtgtaca tttttactgt atttgagaca ttttttgtgt gtgactagtt aattttgcag
     2761 gatgtgccat atcattgaac ggaactaaag tctgtgacag tggatatagc tgctggacca
     2821 ttccatctta tatgttctag cgttctagag cggccgcttc gagcagacat gataagatac
     2881 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2941 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3001 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     3061 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc caggtggcac ttttcgggga
     3121 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3181 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3241 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3301 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3361 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3421 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3481 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3541 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3601 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3661 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3721 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3781 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3841 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3901 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3961 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4021 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4081 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4141 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4201 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4261 tc
//
