LOCUS       sequence-471186-        7466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Study the role and mechanism of the METTL3-P1 3' UTR in regulation 
            gene function..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7466)
  TITLE     Study the role and mechanism of the 3' UTR in regulating gene 
            function.
REFERENCE   2  (bases 1 to 7466)
  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"
COMMENT     Sequence Label: METTL3-P1
FEATURES             Location/Qualifiers
     source          1..7466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(747..1538,1539..1607))
                     /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     1370..1389
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1608..1712)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    1868..1916
                     /note="synthetic polyadenylation signal"
     CDS             complement(1980..2771)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2091..2110)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2701..2720
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(2823..3755)
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     regulatory      3752..3761
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3806..4163)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3820..3955
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3972..4163
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4120..4140
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    4408..4529
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4445..4464)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4499..4518
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(4813..6465)
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     regulatory      6462..6471
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6512..7011)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6560..6579)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     6953..6976
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     misc_feature    7068..7351
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 ttgaaatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct 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 gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccggttg
      721 ggcgtcgctt ggtcggtcat ttcgaattac caatgcttaa tcagtgaggc acctatctca
      781 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
      841 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
      901 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
      961 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1021 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     1081 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     1141 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat agttgtcaga
     1201 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     1261 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     1321 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     1381 ccacatagca gaactttgaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     1441 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     1501 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     1561 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     1621 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     1681 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     1741 gtctaagaaa ccattacgta gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat
     1801 gtcagctact gggctatctg gacaagggaa aacgcaagcg caaagaggat ccttatcgct
     1861 atcgattcac acaaaaaacc aacacacaga tgtaatgaaa ataaagatat tttattgcgg
     1921 ccatcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct
     1981 cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata
     2041 ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg
     2101 gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat
     2161 ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg
     2221 acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg
     2281 agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta
     2341 cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc
     2401 gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga
     2461 gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca
     2521 gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc
     2581 gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc
     2641 gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt
     2701 gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca
     2761 tcttgttcaa tccctccgga gcctccgcca cctgatccgc cacctccgct cccaccaccg
     2821 gtctgctcgt tcttcagcac gcgctccacg aagctcttga tgtacttacc catttcatct
     2881 ggagcgtcct cctggctgaa gtggaggccc ttcaccttca cgaactcggt gttagggaac
     2941 ttcttagctc cctcgacaat agcgttggaa aagaacccag ggtcggactc gatgaacatc
     3001 ttaggcagat cgtcgctggc ccgaaggtag gcgttgtagt tgcggacaat ctggacgacg
     3061 tcgggcttgc ctcccttaac gagagggatc tcgcgaggcc aggagagggt aggccgtcta
     3121 acctcgccct tctccttgaa tggctccagg taggcagcga actcctcagg ctccagtttc
     3181 cgcatgatct tgcttgggag catggtctcg acgaagaagt tattctcaag caccattttc
     3241 tcgccctctt cgctcttgat cagggcgata tcctcctcga tgtcaggcca ctcgtcccag
     3301 gactcgatca cgtccacgac actctcagca tggacgatgg ccttgatctt gtcttggtgc
     3361 tcgtaggagt agtgaaaggc cagacaagcc ccccagtcgt ggcccacaaa gatgattttc
     3421 tttggaaggt tcagcagctc gaaccaagcg gtgaggtact tgtagtgatc caggaggcga
     3481 tatgagccat tcccgctctt gccggactta cccattccga tcagatcagg gatgatgcat
     3541 ctagccacgg gctcgatgtg aggcacgacg tgcctccaca ggtagctgga ggcagcgtta
     3601 ccatgcagaa aaatcacggc gttctcggcg tgcttctcgg aatcatagta gttgatgaag
     3661 gagtccagca cgttcatttg cttgcagcga gcccaccact gaggcccagt gatcatgcgt
     3721 ttgcgttgct cggggtcgta caccttggaa gccatggtgg ctccagcctt tggttcgaga
     3781 ctgttgtgtc agaagaatca agctttttgc aaaagcctag gcctccaaaa aagcctcctc
     3841 actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa
     3901 ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg
     3961 gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc
     4021 ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt
     4081 gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc
     4141 acagctggtt ctttccgcct cagaaggtac ctaaccaagt tcctctttca gaggttattt
     4201 caggccatgg tgctgcgcag atccgcgtat gcggtgtgaa ataccgcaca gatgcgtaag
     4261 gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgattttacc
     4321 acatttgtag aggttttact tgcttaaaaa aacctcccac atctccccct gaacctgaaa
     4381 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     4441 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4501 ggtttgtcca aactcatcaa tgtatcttat catgtctgct cgaagcggcc gccccaaggg
     4561 gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt cgggctttgt
     4621 tagcagccgg atcagcttgc atgcctgcag gtcgactaag cacttcctta cagagctaag
     4681 aatccatagc catggctctg taagctaaac ctgaagagtg atatttgtac aatagctttc
     4741 ttctttattt aaataaacat ttgtattgta gttgggaact cgaggctagc gagctcgttt
     4801 aaacaactag aattacacgg cgatcttgcc gcccttcttg gccttaatga gaatctcgcg
     4861 gatcttgcgg gcgtccaact tgccggtcag tcctttaggc acctcgtcca cgaacacaac
     4921 accaccgcgc agcttcttgg cggttgtaac ctggctggcc acatagtcca cgatctcctt
     4981 ctcggtcatg gttttaccgt gttccagcac gacgactgcg gcgggcagct cgccggcatc
     5041 gtcgtcgggc aggccggcga ccccggcgtc gaagatgttg gggtgttgca gcaggatgct
     5101 ctccagttcg gctggggcta cctggtagcc cttgtatttg atcaggctct tcagccggtc
     5161 cacgatgaag aagtgctcgt cctcgtccca gtaggcgatg tcgccgctgt gcagccagcc
     5221 gtccttgtcg atgagagcgt ttgtagcctc ggggttgtta acgtagccgc tcatgatcat
     5281 ggggccacgg acgcacagct cgccgcgctg gttcacaccc agtgtcttac cggtgtccaa
     5341 gtccaccacc ttagcctcga agaagggcac caccttgcct actgcgccag gcttgtcgtc
     5401 cccttcgggg gtgatcagaa tggcgctggt tgtttctgtc aggccgtagc cctggcggat
     5461 gcctggtagg tggaagcgtt tggccacggc ctcacctacc tccttgctga gcggcgcccc
     5521 gccgctggcg atctcgtgca agttgcttag gtcgtacttg tcgatgagag tgctcttagc
     5581 gaagaagcta aatagtgtgg gcaccagcag ggcagattga atcttatagt cttgcaagct
     5641 gcgcaagaat agctcctcct cgaagcggta catgagcacg acccgaaagc cgcagatcaa
     5701 gtagcccagc gtggtgaaca tgccgaagcc gtggtgaaat ggcaccacgc tgaggatagc
     5761 ggtgtcgggg atgatctggt tgccgaagat ggggtcgcgg gcatgactga atcggacaca
     5821 agcggtgcgg tgcggtaggg ctacgccctt gggcaatccg gtactgccac tactgttcat
     5881 gatcagggcg atggttttgt cccggtcgaa gctctcgggc acgaagtcgt actcgttgaa
     5941 gccgggtggc aaatgggaag tcacgaaggt gtacatgctt tggaagccct ggtagtcggt
     6001 cttgctatcc atgatgatga tcttttgtat gatcggtagc ttcttttgca cgttgaggat
     6061 cttttgcagc cctttcttgc tcacgaatac gacggtgggc tggctgatgc ccatgctgtt
     6121 cagcagctcg cgctcgttgt agatgtcgtt agctggggcc acagccacac cgatgaacag
     6181 ggcacccaac acgggcatga agaactgcaa gctattctcg ctgcacacca cgatccgatg
     6241 gtttgtattc agcccatagc gcttcatagc ttctgccagc cgaacgctca tctcgaagta
     6301 ctcggcgtag gtaatgtcca cctcgatatg tgcgtcggta aaggcgatgg tgccgggcac
     6361 cagggcgtag cgcttcatgg ctttgtgcag ctgctcgccg gcggtcccgt cttcgagtgg
     6421 gtagaatggc gctgggccct tcttaatgtt tttggcatct tccatggtgg ctttaccaac
     6481 agtaccggat tgccaagctt gggctgcagg tcgaaaggcc cggagatgag gaagaggaga
     6541 acagcgcggc agacgtgcgc ttttgaagcg tgcagaatgc cgggcctccg gaggaccttc
     6601 gggcgcccgc cccgcccctg agcccgcccc tgagcccgcc cccggaccca ccccttccca
     6661 gcctctgagc ccagaaagcg aaggagcaaa gctgctattg gccgctgccc caaaggccta
     6721 cccgcttcca ttgctcagcg gtgctgtcca tctgcacgag actagtgaga cgtgctactt
     6781 ccatttgtca cgtcctgcac gacgcgagct gcggggcggg ggggaacttc ctgactaggg
     6841 gaggagtaga aggtggcgcg aaggggccac caaagaacgg agccggttgg cgcctaccgg
     6901 tggatgtgga atgtgtgcga ggccagaggc cacttgtgta gcgccaagtg cccagcgggg
     6961 ctgctaaagc gcatgctcca gactgccttg ggaaaagcgc ctcccctacc cggtagatct
     7021 cgatcctcta cgccggacgc atcgtggccg gagtactgcg gtttatctcg atggctacga
     7081 gggcagacag taagtggatt taccataatc ccttaattgt acgcaccgct aaaacgcgtt
     7141 cagcgcgatc acggcagcag acaggtaaaa atggcaacaa accacccgaa aaactgccgc
     7201 aatcgcgcct gataaatttt aaccgtatga atacctatgc aaccagaggg tacaggccac
     7261 attaccccca cttaatccac tgaagctgcc atttttcatg gtttcaccat cccagcgaag
     7321 ggccatccag cgtgcgttcc tgtatttccg gactataaaa ggatctaggt gaagatcctt
     7381 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     7441 cccgtagaaa agatcaaagg atcttc
//
