LOCUS       sequence-146857-        5553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of miR34.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5553)
  AUTHORS   Eichelser C, Stuckrath I, Muller V, Milde-Langosch K, Wikman H, 
            Pantel K, Schwarzenbach H
  TITLE     Increased serum levels of circulating exosomal microRNA-373 in 
            receptor-negative breast cancer patients.
  JOURNAL   Oncotarget. 2014 Oct 30;5(20):9650-63.
  PUBMED    25333260
REFERENCE   2  (bases 1 to 5553)
  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: 
            "Oncotarget."; date: "2014-10-30"; volume: "5(20)"; pages: "9650-63"
COMMENT     Sequence Label: miR34a in pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..5553
                     /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"
     primer_bind     complement(1147..1164)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1153..1377
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1423..1851
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1510..1529)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1720..1741
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1860..1880)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1865..2194
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2045..2180
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2107..2126
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2261..3055
                     /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(2315..2334)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2925..2944
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3229..3350
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3266..3285)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3320..3339
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3399..3415)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3399..3415)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3412..3434)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3423..3439
                     /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(3447..3453,3454..3471,3472..3477))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3492..3513
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3630..3647)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3801..4386)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3881..3900)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4557..5348,5349..5417))
                     /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     5180..5199
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5418..5522)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc
      961 agatatccag cacagtggcg gccgctcgag atctggccag ctgtgagtgt ttctttggca
     1021 gtgtcttagc tggttgttgt gagcaatagt aaggaagcaa tcagcaagta tactgcccta
     1081 gaagtgctgc acgttgtggg gcccggatcc tcgagtctag agggcccgtt taaacccgct
     1141 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     1201 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     1261 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     1321 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     1381 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     1441 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     1501 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     1561 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     1621 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     1681 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     1741 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     1801 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     1861 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     1921 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     1981 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     2041 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     2101 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     2161 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     2221 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     2281 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     2341 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     2401 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     2461 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     2521 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     2581 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     2641 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     2701 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     2761 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     2821 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     2881 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     2941 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3001 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     3061 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     3121 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     3181 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     3241 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3301 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     3361 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     3421 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3481 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3541 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3601 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3661 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3721 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3781 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3841 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3901 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3961 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4021 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4081 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4141 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4201 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4261 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4321 accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4381 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4441 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4501 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4561 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4621 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4681 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4741 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4801 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4861 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4921 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4981 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5041 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5101 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5161 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5221 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5281 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5341 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5401 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5461 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5521 cgcacatttc cccgaaaagt gccacctgac gtc
//
