LOCUS       sequence-7120-ex        5367 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 5367)
  AUTHORS   Lewis BP, Shih IH, Jones-Rhoades MW, Bartel DP, Burge CB
  TITLE     Prediction of mammalian microRNA targets.
  JOURNAL   Cell. 2003 Dec 26. 115(7):787-98.
  PUBMED    14697198
REFERENCE   2  (bases 1 to 5367)
  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: "Cell. 2003 
            Dec 26. 115(7):787-98."
COMMENT     Sequence Label: pISlin41m(s) (lin-41 microRNA mut reporter)
FEATURES             Location/Qualifiers
     source          1..5367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..212
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      63..198
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      242..251
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             248..1900
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(314..332)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2084..2205
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2095..2114)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2149..2168
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2527..2547
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(2699..2716)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2870..3458)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2950..2969)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3629..4420,4421..4489))
                     /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     4252..4271
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4490..4594)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4621..5076
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4708..4727)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4918..4939
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5207..5255
                     /note="synthetic polyadenylation signal"
     misc_feature    5269..5360
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5309..5328
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtagatctg cgatctgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
       61 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatcgc tgactaattt
      121 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
      181 gaggcttttt tggaggccta ggcttttgca aaaagcttgg cattccggta ctgttggtaa
      241 agccaccatg gaagacgcca aaaacataaa gaaaggcccg gcgccattct atccgctgga
      301 agatggaacc gctggagagc aactgcataa ggctatgaag agatacgccc tggttcctgg
      361 aacaattgct tttacagatg cacatatcga ggtggacatc acttacgctg agtacttcga
      421 aatgtccgtt cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat
      481 cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat
      541 cggagttgca gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat
      601 gggcatttcg cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa
      661 cgtgcaaaaa aagctcccaa tcatccaaaa aattattatc atggattcta aaacggatta
      721 ccagggattt cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga
      781 atacgatttt gtgccagagt ccttcgatag ggacaagaca attgcactga tcatgaactc
      841 ctctggatct actggtctgc ctaaaggtgt cgctctgcct catagaactg cctgcgtgag
      901 attctcgcat gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt
      961 aagtgttgtt ccattccatc acggttttgg aatgtttact acactcggat atttgatatg
     1021 tggatttcga gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga ggagccttca
     1081 ggattacaag attcaaagtg cgctgctggt gccaacccta ttctccttct tcgccaaaag
     1141 cactctgatt gacaaatacg atttatctaa tttacacgaa attgcttctg gtggcgctcc
     1201 cctctctaag gaagtcgggg aagcggttgc caagaggttc catctgccag gtatcaggca
     1261 aggatatggg ctcactgaga ctacatcagc tattctgatt acacccgagg gggatgataa
     1321 accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg atctggatac
     1381 cgggaaaacg ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc ctatgattat
     1441 gtccggttat gtaaacaatc cggaagcgac caacgccttg attgacaagg atggatggct
     1501 acattctgga gacatagctt actgggacga agacgaacac ttcttcatcg ttgaccgcct
     1561 gaagtctctg attaagtaca aaggctatca ggtggctccc gctgaattgg aatccatctt
     1621 gctccaacac cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg
     1681 tgaacttccc gccgccgttg ttgttttgga gcacggaaag acgatgacgg aaaaagagat
     1741 cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt
     1801 tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat
     1861 cctcataaag gccaagaagg gcggaaagat cgccgtgtaa ttctaggagc tctttcctca
     1921 aattgcacca actcaagtat accttttata catccgttct caactcaacg cgatgtaaat
     1981 atcgcaatcc ctttttatac atccattctc gctctgaacc attgaaacct tctccctcta
     2041 gcgttctaga gtcggggcgg ccggccgctt cgagcagaca tgataagata cattgatgag
     2101 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     2161 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     2221 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     2281 ctctacaaat gtggtaaaat cgataaggat ctgaacgatg gagcggagaa tgggcggaac
     2341 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
     2401 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
     2461 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
     2521 ttccacaccc taactgacac acattccaca gcggatccgt cgaccgatgc ccttgagagc
     2581 cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat
     2641 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct tccgcttcct
     2701 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2761 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2821 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2881 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2941 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3001 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3061 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3121 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3181 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3241 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3301 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3361 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3421 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3481 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3541 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3601 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3661 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3721 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3781 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3841 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3901 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3961 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4021 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4081 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4141 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4201 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4261 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4321 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4381 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4441 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4501 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4561 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4621 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4681 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4741 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4801 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4861 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4921 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4981 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     5041 acgcgaattt taacaaaata ttaacgctta caatttgcca ttcgccattc aggctgcgca
     5101 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagccc aagctaccat
     5161 gataagtaag taatattaag gtacgggagg tacttggagc ggccgcaata aaatatcttt
     5221 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagtactaa catacgctct
     5281 ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc
     5341 aggtgccaga acatttctct atcgata
//
