LOCUS       sequence-78273-e        6532 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 6532)
  AUTHORS   Park SY, Lee JH, Ha M, Nam JW, Kim VN
  TITLE     miR-29 miRNAs activate p53 by targeting p85 alpha and CDC42.
  JOURNAL   Nat Struct Mol Biol. 2009 Jan;16(1):23-9. doi: 10.1038/nsmb.1533. 
            Epub 2008 Dec 14.
  PUBMED    19079265
REFERENCE   2  (bases 1 to 6532)
  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: "10.1038/nsmb.1533";
            journalName: "Nat Struct Mol Biol"; date: "2009-01"; volume: "16"; 
            issue: "1"; pages: "23-9"
COMMENT     Sequence Label: pcDNA3-miR-29b-1~29a
FEATURES             Location/Qualifiers
     source          1..6532
                     /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(2084..2102)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2085..2102)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2122..2139)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2128..2352
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2398..2826
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2485..2504)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2695..2716
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2835..2855)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2840..3170
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3021..3156
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3083..3102
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3237..4031
                     /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(3291..3310)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3901..3920
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4205..4326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4242..4261)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4296..4315
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4375..4391)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4375..4391)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4388..4410)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4399..4415
                     /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(4423..4429,4430..4447,4448..4453))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4468..4489
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4606..4623)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4777..5365)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4857..4876)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5536..6327,6328..6396))
                     /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     6159..6178
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6397..6501)
                     /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 ccactagtaa cggccgccag tgtgctggaa ttcctctccc atcaataaca
      961 attcagtgac atcagtttat gaatatatga aatttgccaa agctctgttt agaccactga
     1021 gtaactcaca gctaggtttc aacttttcct ttctaggttg tcttgggttt attgtaagag
     1081 agcattatga agaaaaaaat agatcataaa gcttcttcag gaagctggtt tcatatggtg
     1141 gtttagattt aaatagtgat tgtctagcac catttgaaat cagtgttctt gggggagacc
     1201 agctgcgctg cactaccaac agcaaaagaa gtgaatggga cagctctgaa gtatttgaaa
     1261 gcaacagcag gatggctgtg agaacctgcc tcacatgtag ctgacccctt cctcacccct
     1321 gccaacagtg gtggcatata tcacaaatgg cagtcaggtc tctgcactgg cggatccaac
     1381 tgtgatcgaa agttttccaa aaataagttg tgtctgtatt gaacatgaac agactttctt
     1441 cttgtcatta ttctctaaca atactgcata acaattattt gcatacattt gcattgcatt
     1501 aagtattcta agtaatctag agacgattta aagtatacgg gaggatgtgt gtaggttgta
     1561 tgcaaatact acaccatttc tatcagagac ttgagcatct gtggattttg gtatccaagg
     1621 ggctttctgg aaccaatccc tcaaggatac caagggatga atgtaattgt acaggatatc
     1681 gcattgttgg aattttatac ttctttgtgg aataaaccta tagcacttaa tagatagtac
     1741 agactcattc cattgtgcct gggttaaaga gcccaatgta tgctggattt agtaagattt
     1801 gggccctccc aaccctcacg accttctgtg accccttaga ggatgactga tttcttttgg
     1861 tgttcagagt caatataatt ttctagcacc atctgaaatc ggttataatg attggggaag
     1921 agcaccatga tgctgactgc tgagaggaaa tgtattggtg accgttgggg ccatggacaa
     1981 gaactaagaa aacaaatgca aagcaataat gcaaaggtga tttttcttct tccagtttct
     2041 aagttgaatt tcactgctcg agcatgcatc tagagggccc tattctatag tgtcacctaa
     2101 atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     2161 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2221 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2281 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     2341 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     2401 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2461 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2521 tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg
     2581 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     2641 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2701 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     2761 ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2821 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagg
     2881 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2941 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3001 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3061 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3121 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3181 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3241 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3301 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3361 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3421 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3481 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3541 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3601 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3661 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3721 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3781 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3841 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3901 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3961 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4021 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4081 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4141 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4201 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4261 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4321 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4381 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4441 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4501 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4561 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4621 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4681 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4741 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4801 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4861 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4921 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     4981 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5041 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5101 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5161 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     5221 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5281 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5341 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5401 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5461 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5521 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5581 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5641 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5701 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5761 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5821 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5881 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5941 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6001 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6061 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6121 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6181 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6241 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6301 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6361 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6421 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6481 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
