LOCUS       sequence-273256-        6907 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 6907)
  AUTHORS   Ogunjimi AA, Wiesner S, Briant DJ, Varelas X, Sicheri F, Forman-Kay 
            J, Wrana JL
  TITLE     The ubiquitin binding region of the Smurf HECT domain facilitates 
            polyubiquitylation and binding of ubiquitylated substrates.
  JOURNAL   J Biol Chem. 2010 Feb 26. 285(9):6308-15.
  PUBMED    20026602
REFERENCE   2  (bases 1 to 6907)
  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: "J Biol 
            Chem. 2010 Feb 26. 285(9):6308-15."
COMMENT     Sequence Label: pCMV5 MYC-Smurf2 (FF29/30A) WT
FEATURES             Location/Qualifiers
     source          1..6907
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             636..665
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    2931..3553
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3130..3149)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3577..3597)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3582..3911
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3762..3897
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3949..3968)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3950..3968)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3982..3998)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3982..3998)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3995..4017)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4006..4022
                     /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(4030..4036,4037..4054,4055..4060))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4075..4096
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4213..4230)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4384..4972)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4464..4483)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5143..5934,5935..6003))
                     /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     5766..5785
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6004..6108)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6135..6590
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6222..6241)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6432..6453
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6716..6738
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6730..6747
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6731..6747
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattgatc tggtaccacg cgtatcgata ccatggagca gaagctgata tccgaggagg
      661 acctaggtcg acaatctaac cccggacgcc ggaggaacgg gcccgtcaag ctgcgcctga
      721 cagtactctg tgcaaaaaac ctggtgaaaa aggatgctgc ccgacttcct gatccatttg
      781 ctaaggtggt ggttgatgga tctgggcaat gccattctac agatactgtg aagaatacgc
      841 ttgatccaaa gtggaatcag cattatgacc tgtatattgg aaagtctgat tcagttacga
      901 tcagtgtatg gaatcacaag aagatccata agaaacaagg tgctggattt ctcggttgtg
      961 ttcgtcttct ttccaatgcc atcaaccgcc tcaaagacac tggttatcag aggttggatt
     1021 tatgcaaact cgggccaaat gacaatgata cagttagagg acagatagta gtaagtcttc
     1081 agtccagaga ccgaataggc acaggaggac aagttgtgga ctgcagtcgt ttatttgata
     1141 acgatttacc agacggctgg gaagaaagga gaaccgcctc tggaagaatc cagtatctaa
     1201 accatataac aagaactacg caatgggagc gcccaacacg accggcatcc gaatattcta
     1261 gccctggcag acctcttagc tgctttgttg atgagaacac tccaattagt ggaacaaatg
     1321 gtgcaacatg tggacagtct tcagatccca ggctggcaga gaggagagtc aggtcacaac
     1381 gacatagaaa ttacatgagc agaacacatt tacatactcc tccagaccta ccagaaggct
     1441 atgaacagag gacaacgcaa caaggccagg tgtatttctt acatacacag actggtgtga
     1501 gcacatggca tgatccaaga gtgcccaggg atcttagcaa catcaattgt gaagagcttg
     1561 gtccattgcc tcctggatgg gagatccgta atacggcaac aggcagagtt tatttcgttg
     1621 accataacaa cagaacaaca caatttacgg atcctcggct gtctgctaac ttgcatttag
     1681 ttttaaatcg gcagaaccaa ttgaaagacc aacagcaaca gcaagtggta tcgttatgtc
     1741 ctgatgacac agaatgcctg acagtcccaa ggtacaagcg agacctggtt cagaaactaa
     1801 aaattttgcg gcaagaactt tcccaacaac agcctcaggc aggtcattgc cgcattgagg
     1861 tttccaggga agagattttt gaggaatcat atcgacaggt catgaaaatg agaccaaaag
     1921 atctctggaa gcgattaatg ataaaatttc gtggagaaga aggccttgac tatggaggcg
     1981 ttgccaggga atggttgtat ctcttgtcac atgaaatgtt gaatccatac tatggcctct
     2041 tccagtattc aagagatgat atttatacat tgcagatcaa tcctgattct gcagttaatc
     2101 cggaacattt atcctatttc cactttgttg gacgaataat gggaatggct gtgtttcatg
     2161 gacattatat tgatggtggt ttcacattgc ctttttataa gcaattgctt gggaagtcaa
     2221 ttaccttgga tgacatggag ttagtagatc cggatcttca caacagttta gtgtggatac
     2281 ttgagaatga tattacaggt gttttggacc ataccttctg tgttgaacat aatgcatatg
     2341 gtgaaattat tcagcatgaa cttaaaccaa atggcaaaag tatccctgtt aatgaagaaa
     2401 ataaaaaaga atatgtcagg ctctatgtga actggagatt tttacgaggc attgaggctc
     2461 aattcttggc tctgcagaaa ggatttaatg aagtaattcc acaacatctg ctgaagacat
     2521 ttgatgagaa ggagttagag ctcattattt gtggacttgg aaagatagat gttaatgact
     2581 ggaaggtaaa cacccggtta aaacactgta caccagacag caacattgtc aaatggttct
     2641 ggaaagctgt ggagtttttt gatgaagagc gacgagcaag attgcttcag tttgtgacag
     2701 gatcctctcg agtgcctctg cagggcttca aagcattgca aggtgctgca ggcccgagac
     2761 tctttaccat acaccagatt gatgcctgca ctaacaacct gccgaaagcc cacacttgct
     2821 tcaatcgaat agacattcca ccctatgaaa gctatgaaaa gctatatgaa aagctgctaa
     2881 cagccattga agaaacatgt ggatttgctg tggaatgaca atctagaccc gggtggcatc
     2941 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca
     3001 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata
     3061 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
     3121 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa
     3181 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc
     3241 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca
     3301 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca
     3361 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttaaaat
     3421 aactatacca gcaggaggac gtccagacac agcataggct acctggccat gcccaaccgg
     3481 tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc actcagtaga
     3541 tgcctgttga attgggtacg cggccagctt ggctgtggaa tgtgtgtcag ttagggtgtg
     3601 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3661 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3721 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3781 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3841 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3901 gcttttgcaa aaagctcctc gaggaactga aaaaccagaa agttaattcc ctatagtgag
     3961 tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4021 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4081 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4141 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4201 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4261 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4321 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4381 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4441 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4501 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4561 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4621 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4681 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4741 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4801 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4861 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4921 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4981 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5041 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5101 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5161 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5221 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5281 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5341 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5401 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5461 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5521 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5581 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5641 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5701 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5761 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5821 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5881 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5941 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6001 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6061 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6121 aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     6181 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     6241 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     6301 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     6361 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     6421 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     6481 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     6541 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca
     6601 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     6661 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     6721 tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa tcaatattgg
     6781 caattagcca tattagtcat tggttatata gcataaatca atattggcta ttggccattg
     6841 catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg
     6901 ccatgtt
//
