LOCUS       sequence-78275-e        5613 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 5613)
  AUTHORS   Heo I, Joo C, Cho J, Ha M, Han J, Kim VN
  TITLE     Lin28 mediates the terminal uridylation of let-7 precursor MicroRNA.
  JOURNAL   Mol Cell. 2008 Oct 24;32(2):276-84. doi: 
            10.1016/j.molcel.2008.09.014.
  PUBMED    18951094
REFERENCE   2  (bases 1 to 5613)
  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"; doi: 
            "10.1016/j.molcel.2008.09"; journalName: "Mol Cell"; date: 
            "2008-10-24- 24"; volume: "32"; issue: "2"; pages: "276-84"
COMMENT     Sequence Label: pcDNA3-pri-let-7b
FEATURES             Location/Qualifiers
     source          1..5613
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1165..1183)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1166..1183)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1203..1220)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1209..1433
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1479..1907
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1566..1585)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1776..1797
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1916..1936)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1921..2251
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2102..2237
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2164..2183
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2318..3112
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2372..2391)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2982..3001
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3286..3407
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3323..3342)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3377..3396
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3456..3472)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3456..3472)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3469..3491)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3480..3496
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3504..3510,3511..3528,3529..3534))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3549..3570
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3687..3704)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3858..4446)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3938..3957)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4617..5408,5409..5477))
                     /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     5240..5259
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5478..5582)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cctgctctgg tgactgagga ccgccaggca ggggctggtg ctgggcgggg
      961 ggcggcgggc cctcccgcag tgcaaggccg ggcctggcgg ggtgaggtag taggttgtgt
     1021 ggtttcaggg cagtgatgtt gcccctcgga agataactat acaacctact gccttccctg
     1081 aggagcccag tgacacgacc ccatgggagg gccgccccct acctcagtga catgaccctc
     1141 gagcatgcat ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc
     1201 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     1261 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     1321 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     1381 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     1441 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     1501 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     1561 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     1621 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     1681 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     1741 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     1801 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     1861 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     1921 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag gcaggcagaa gtatgcaaag
     1981 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2041 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2101 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2161 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     2221 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     2281 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     2341 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     2401 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     2461 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     2521 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     2581 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     2641 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     2701 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     2761 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     2821 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     2881 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     2941 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     3001 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     3061 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     3121 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     3181 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     3241 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     3301 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3361 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     3421 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     3481 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     3541 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     3601 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3661 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3721 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3781 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3841 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3901 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3961 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4021 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     4081 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4141 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4201 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4261 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     4321 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4381 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4441 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4501 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4561 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4621 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4681 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4741 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4801 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4861 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4921 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4981 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5041 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5101 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5161 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5221 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5281 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5341 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5401 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5461 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5521 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5581 cgcacatttc cccgaaaagt gccacctgac gtc
//
