LOCUS       sequence-424377-        6884 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 6884)
  AUTHORS   Yamashita M, Ying SX, Zhang GM, Li C, Cheng SY, Deng CX, Zhang YE
  TITLE     Ubiquitin ligase Smurf1 controls osteoblast activity and bone 
            homeostasis by targeting MEKK2 for degradation.
  JOURNAL   Cell. 2005 Apr 8. 121(1):101-13.
  PUBMED    15820682
REFERENCE   2  (bases 1 to 6884)
  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. 2005 
            Apr 8. 121(1):101-13."
COMMENT     Sequence Label: pRK-Myc-Smurf1
FEATURES             Location/Qualifiers
     source          1..6884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(21..40)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     75..94
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        187..544
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        187..378
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(210..230)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      395..530
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(564..581)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(564..580)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(573..595)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      793..1248
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(880..899)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1090..1111
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1265..1284)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1384..1406
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1444..1462)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1530..1634
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1635..1703,1704..2495)
                     /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     complement(1853..1872)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2666..3254
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3155..3174
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3408..3425
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3542..3563
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3578..3583,3584..3601,3602..3608)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3616..3632
                     /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)."
     primer_bind     3621..3643
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3640..3656
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3640..3656
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        3685..4064
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4065..4268
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4219..4239
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4265..4289
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        4499..4517
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4499..4516
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     4530..4549
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      4589..4598
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4598..4627
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      6855..6864
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    join(6868..6884,1..118)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
       61 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcgg
      121 gaattaattc ggcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg
      181 taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
      241 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
      301 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
      361 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
      421 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
      481 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
      541 caaaaagctg ttaacagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac
      601 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
      661 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
      721 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
      781 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
      841 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
      901 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
      961 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     1021 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     1081 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt
     1141 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     1201 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     1261 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     1321 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     1381 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     1441 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     1501 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1561 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1621 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1681 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1741 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1801 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1861 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1921 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     1981 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2041 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2101 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2161 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     2221 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     2281 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2341 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2401 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2461 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2521 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2581 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2641 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2701 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2761 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     2821 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2881 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2941 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3001 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3061 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3121 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3181 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3241 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3301 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3361 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3421 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     3481 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     3541 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     3601 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga
     3661 ttacgaatta attcgagctc gcccgacatt gattattgac tagttattaa tagtaatcaa
     3721 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     3781 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     3841 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     3901 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     3961 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     4021 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     4081 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     4141 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     4201 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     4261 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     4321 tccatagaag acaccgggac cgatccagcc tccgcggccg ggaacggtgc attggaacgc
     4381 ggattccccg tgccaagagt gacgtaagta ccgcctatag agtctatagg cccaccccct
     4441 tggcttcgtt agaacgcggc tacaattaat acataacctt atgtatcata cacatacgat
     4501 ttaggtgaca ctatagaata acatccactt tgcctttctc tccacaggtg tccactccca
     4561 ggtccaactg cacctcggtt ctatcgccgc caccatggag cagaagctga tatcagagga
     4621 ggatctgatc gatgaattca tgtcgaaccc cgggacacgc aggaacggct ccagcatcaa
     4681 gatccgtctg acagtgttat gtgccaagaa ccttgcaaag aaagacttct tcaggctccc
     4741 tgaccctttt gcaaagattg tcgtggatgg gtctgggcag tgccactcaa ccgacactgt
     4801 gaaaaacaca ttggacccaa agtggaacca gcactatgat ctatatgttg ggaaaacgga
     4861 ttcgataacc attagcgtgt ggaaccataa gaaaattcac aagaaacagg gagctggctt
     4921 cctgggctgt gtgcggctgc tctccaatgc catcagcaga ttaaaagata ccggatacca
     4981 gcgtttggat ctatgcaaac taaacccctc agatactgat gcagttcgtg gccagatagt
     5041 ggtcagttta cagacacgag acagaatagg aaccggcggc tcggtggtgg actgcagagg
     5101 actgttagaa aatgaaggaa cggtgtatga agactccggg cctgggaggc cgctcagctg
     5161 cttcatggag gaaccagccc cttacacaga tagcaccggt gctgctgctg gaggagggaa
     5221 ttgcaggttc gtggagtccc caagtcaaga tcaaagactt caggcacagc ggcttcgaaa
     5281 ccctgatgtg cgaggttcac tacagacgcc ccagaaccga ccacacggcc accagtcccc
     5341 ggaactgccc gaaggctacg aacaaagaac aacagtccag ggccaagttt actttttgca
     5401 tacacagact ggagttagca cgtggcacga ccccaggata ccaagagacc ttaacagtgt
     5461 gaactgtgat gaacttggac cactgccgcc aggctgggaa gtcagaagta cagtttctgg
     5521 gaggatatat tttgtagatc ataataaccg aacaacccag tttacagacc caaggttaca
     5581 ccacatcatg aatcaccagt gccaactcaa ggagcccagc cagccgctgc cactgcccag
     5641 tgagggctct ctggaggacg aggagcttcc tgcccagaga tacgaaagag atctagtcca
     5701 gaagctgaaa gtcctcagac acgaactgtc gcttcagcag ccccaagctg gtcattgccg
     5761 catcgaagtg tccagagaag aaatctttga ggagtcttac cgccagataa tgaagatgcg
     5821 accgaaagac ttgaaaaaac ggctgatggt gaaattccgt ggggaagaag gtttggatta
     5881 cggtggtgtg gccagggagt ggctttactt gctgtgccat gaaatgctga atccttatta
     5941 cgggctcttc cagtattcta cggacaatat ttacatgttg caaataaatc cggattcttc
     6001 aatcaacccc gaccacttgt cttatttcca ctttgtgggg cggatcatgg ggctggctgt
     6061 gttccatgga cactacatca acgggggctt cacagtgccc ttctacaagc agctgctggg
     6121 gaagcccatc cagctctcag atctggaatc tgtggaccca gagctgcata agagcttggt
     6181 gtggatccta gagaacgaca tcacgcctgt actggaccac accttctgcg tggaacacaa
     6241 cgccttcggg cggatcctgc agcatgaact gaaacccaat ggcagaaatg tgccagtcac
     6301 agaggagaat aagaaagaat acgtccggtt gtatgtaaac tggaggttta tgagaggaat
     6361 cgaagcccag ttcttagctc tgcagaaggg gttcaatgag ctcatccctc aacatctgct
     6421 gaagcctttt gaccagaagg aactggagct gatcataggc ggcctggata aaatagactt
     6481 gaacgactgg aagtcgaaca cgcggctgaa gcactgtgtg gccgacagca acatcgtgcg
     6541 gtggttctgg caagcggtgg agacgttcga tgaagaaagg agggccaggc tcctgcagtt
     6601 tgtgactggg tccacgcgag tcccgctcca aggcttcaag gctttgcaag gttctacagg
     6661 cgcggcaggg ccccggctgt tcaccatcca cctgatagac gcgaacacag acaaccttcc
     6721 gaaggcccat acctgcttta accggatcga cattccacca tatgagtcct atgagaagct
     6781 ctacgagaag ctgctgacag ccgtggagga gacctgcggg tttgctgtgg agtaagtcga
     6841 cctgcagaag cttggccgcc atggcccaac ttgtttattg cagc
//
