LOCUS       sequence-235250-        4568 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 4568)
  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 4568)
  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: pIS47 C20orf139 3'UTR mut
FEATURES             Location/Qualifiers
     source          1..4568
                     /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    3179..3300
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3190..3209)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3244..3263
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3433..3537
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3538..3606,3607..4398)
                     /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(3756..3775)
                     /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 ggagctctgt
     2641 ctgcactgct ggtatggggt taggccaggt aggaaagtac agaggggctt ctgaaaacca
     2701 agagtccctg gggaaaggga acagagtaag gcaggccttg ttctcactgc cctctaaggg
     2761 aacttggtca ctcggcactt ttaagcctca gtttctccag ttcaataata aggacaagag
     2821 cttttcccat gcattctctt tccccgggaa agttgactga ggtgaccagt aatagaattg
     2881 aaaagggaga gtgtcttcag tgcaatgtgg catcctggat tgggtcttgg aacaaaaaca
     2941 ggacattagt gggaaaattg gaaatctgaa aaaagtctga attttagtta atataccaat
     3001 ttcagtctct tggttttgac agatgtacca tggtgatgta agatgttgac cttggggtag
     3061 gctgggtgaa gggtatacag gaactctttg tactatctct gcaacttctc tgtaaatcta
     3121 gtataagtac aaaataaaag ttctagcgtt ctagagcggc cgcttcgagc agacatgata
     3181 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     3241 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     3301 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     3361 taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccagg tggcactttt
     3421 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3481 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3541 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3601 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3661 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3721 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3781 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3841 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3901 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3961 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4021 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4081 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4141 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4201 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4261 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4321 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4381 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4441 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4501 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4561 ggatcttc
//
