LOCUS       sequence-239412-        6880 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 6880)
  AUTHORS   Kavsak P, Rasmussen RK, Causing CG, Bonni S, Zhu H, Thomsen GH, 
            Wrana JL
  TITLE     Smad7 binds to Smurf2 to form an E3 ubiquitin ligase that targets 
            the TGF beta receptor for degradation.
  JOURNAL   Mol Cell. 2000 Dec . 6(6):1365-75.
  PUBMED    11163210
REFERENCE   2  (bases 1 to 6880)
  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: "Mol Cell. 
            2000 Dec . 6(6):1365-75."
COMMENT     Sequence Label: pCMV5B-Flag-Smurf2 C716A
FEATURES             Location/Qualifiers
     source          1..6880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(82..102)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      247..381
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(433..452)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(434..452)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(466..482)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(466..482)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(479..501)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    490..506
                     /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(514..520,521..538,539..544))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    559..580
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(697..714)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(868..1456)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(948..967)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1627..2418,2419..2487))
                     /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     2250..2269
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2488..2592)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2619..3074
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2706..2725)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2916..2937
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3200..3222
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3214..3231
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3215..3231
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3392..3771
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3772..3975
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3926..3946
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4027..4050
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    join(6316..6880,1..58)
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(6515..6534)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
ORIGIN
        1 gcttgcttgg cactgtcctc tcatgcgttg ggtccactca gtagatgcct gttgaattgg
       61 gtacgcggcc agcttggctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
      121 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
      181 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
      241 ccatagtccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
      301 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
      361 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cctcgaggaa ctgaaaaacc
      421 agaaagttaa ttccctatag tgagtcgtat taaattcgta atcatgtcat agctgtttcc
      481 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
      541 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
      601 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
      661 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
      721 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
      781 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
      841 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
      901 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
      961 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     1021 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     1081 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     1141 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     1201 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     1261 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     1321 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     1381 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     1441 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     1501 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     1561 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1621 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1681 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1741 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     1801 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1861 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1921 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1981 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     2041 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     2101 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     2161 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     2221 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     2281 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     2341 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     2401 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     2461 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     2521 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     2581 aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt
     2641 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2701 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2761 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2821 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2881 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2941 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3001 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3061 aacgcttaca atttccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     3121 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     3181 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
     3241 atctatacat tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa
     3301 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat
     3361 attggctcat gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag
     3421 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     3481 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     3541 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     3601 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
     3661 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
     3721 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     3781 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     3841 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     3901 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     3961 tatataagca gagctcgttt agtgaaccgt cagaattgat ctggtaccac gcgtatcgat
     4021 accatggact acaaggacga cgatgacaag ggtcgacaat ctaaccccgg acgccggagg
     4081 aacgggcccg tcaagctgcg cctgacagta ctctgtgcaa aaaacctggt gaaaaaggat
     4141 tttttccgac ttcctgatcc atttgctaag gtggtggttg atggatctgg gcaatgccat
     4201 tctacagata ctgtgaagaa tacgcttgat ccaaagtgga atcagcatta tgacctgtat
     4261 attggaaagt ctgattcagt tacgatcagt gtatggaatc acaagaagat ccataagaaa
     4321 caaggtgctg gatttctcgg ttgtgttcgt cttctttcca atgccatcaa ccgcctcaaa
     4381 gacactggtt atcagaggtt ggatttatgc aaactcgggc caaatgacaa tgatacagtt
     4441 agaggacaga tagtagtaag tcttcagtcc agagaccgaa taggcacagg aggacaagtt
     4501 gtggactgca gtcgtttatt tgataacgat ttaccagacg gctgggaaga aaggagaacc
     4561 gcctctggaa gaatccagta tctaaaccat ataacaagaa ctacgcaatg ggagcgccca
     4621 acacgaccgg catccgaata ttctagccct ggcagacctc ttagctgctt tgttgatgag
     4681 aacactccaa ttagtggaac aaatggtgca acatgtggac agtcttcaga tcccaggctg
     4741 gcagagagga gagtcaggtc acaacgacat agaaattaca tgagcagaac acatttacat
     4801 actcctccag acctaccaga aggctatgaa cagaggacaa cgcaacaagg ccaggtgtat
     4861 ttcttacata cacagactgg tgtgagcaca tggcatgatc caagagtgcc cagggatctt
     4921 agcaacatca attgtgaaga gcttggtcca ttgcctcctg gatgggagat ccgtaatacg
     4981 gcaacaggca gagtttattt cgttgaccat aacaacagaa caacacaatt tacggatcct
     5041 cggctgtctg ctaacttgca tttagtttta aatcggcaga accaattgaa agaccaacag
     5101 caacagcaag tggtatcgtt atgtcctgat gacacagaat gcctgacagt cccaaggtac
     5161 aagcgagacc tggttcagaa actaaaaatt ttgcggcaag aactttccca acaacagcct
     5221 caggcaggtc attgccgcat tgaggtttcc agggaagaga tttttgagga atcatatcga
     5281 caggtcatga aaatgagacc aaaagatctc tggaagcgat taatgataaa atttcgtgga
     5341 gaagaaggcc ttgactatgg aggcgttgcc agggaatggt tgtatctctt gtcacatgaa
     5401 atgttgaatc catactatgg cctcttccag tattcaagag atgatattta tacattgcag
     5461 atcaatcctg attctgcagt taatccggaa catttatcct atttccactt tgttggacga
     5521 ataatgggaa tggctgtgtt tcatggacat tatattgatg gtggtttcac attgcctttt
     5581 tataagcaat tgcttgggaa gtcaattacc ttggatgaca tggagttagt agatccggat
     5641 cttcacaaca gtttagtgtg gatacttgag aatgatatta caggtgtttt ggaccatacc
     5701 ttctgtgttg aacataatgc atatggtgaa attattcagc atgaacttaa accaaatggc
     5761 aaaagtatcc ctgttaatga agaaaataaa aaagaatatg tcaggctcta tgtgaactgg
     5821 agatttttac gaggcattga ggctcaattc ttggctctgc agaaaggatt taatgaagta
     5881 attccacaac atctgctgaa gacatttgat gagaaggagt tagagctcat tatttgtgga
     5941 cttggaaaga tagatgttaa tgactggaag gtaaacaccc ggttaaaaca ctgtacacca
     6001 gacagcaaca ttgtcaaatg gttctggaaa gctgtggagt tttttgatga agagcgacga
     6061 gcaagattgc ttcagtttgt gacaggatcc tctcgagtgc ctctgcaggg cttcaaagca
     6121 ttgcaaggtg ctgcaggccc gagactcttt accatacacc agattgatgc ctgcactaac
     6181 aacctaccga aagcccacac tgccttcaat cgaatagaca ttccacccta tgaaagctat
     6241 gaaaagctat atgaaaagct gctaacagcc attgaagaaa catgtggatt tgctgtggaa
     6301 tgacaatcta gacccgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     6361 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     6421 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     6481 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     6541 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     6601 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     6661 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
     6721 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
     6781 ttccctgtcc ttctgatttt aaaataacta taccagcagg aggacgtcca gacacagcat
     6841 aggctacctg gccatgccca accggtggga catttgagtt
//
