LOCUS       sequence-169234-        6901 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 6901)
  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 6901)
  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 wt
FEATURES             Location/Qualifiers
     source          1..6901
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..114
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     106..123
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     107..123
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        284..663
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        664..867
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     818..838
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             919..942
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3208..3830
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3407..3426)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3854..3874)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3859..4188
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4039..4174
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4226..4245)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4227..4245)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4259..4275)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4259..4275)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4272..4294)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4283..4299
                     /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(4307..4313,4314..4331,4332..4337))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4352..4373
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4490..4507)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4661..5249)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4741..4760)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5420..6211,6212..6280))
                     /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     6043..6062
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6281..6385)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6412..6867
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6499..6518)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6709..6730
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
       61 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
      121 agtgccaagc tgatctatac attgaatcaa tattggcaat tagccatatt agtcattggt
      181 tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat
      241 atgtacattt atattggctc atgtccaata tgaccgccat gttgacattg attattgact
      301 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      361 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      421 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      481 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      541 agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      601 atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      661 atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga
      721 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      781 gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta
      841 cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagaattg atctggtacc
      901 acgcgtatcg ataccatgga ctacaaggac gacgatgaca agggtcgaca atctaacccc
      961 ggacgccgga ggaacgggcc cgtcaagctg cgcctgacag tactctgtgc aaaaaacctg
     1021 gtgaaaaagg attttttccg acttcctgat ccatttgcta aggtggtggt tgatggatct
     1081 gggcaatgcc attctacaga tactgtgaag aatacgcttg atccaaagtg gaatcagcat
     1141 tatgacctgt atattggaaa gtctgattca gttacgatca gtgtatggaa tcacaagaag
     1201 atccataaga aacaaggtgc tggatttctc ggttgtgttc gtcttctttc caatgccatc
     1261 aaccgcctca aagacactgg ttatcagagg ttggatttat gcaaactcgg gccaaatgac
     1321 aatgatacag ttagaggaca gatagtagta agtcttcagt ccagagaccg aataggcaca
     1381 ggaggacaag ttgtggactg cagtcgttta tttgataacg atttaccaga cggctgggaa
     1441 gaaaggagaa ccgcctctgg aagaatccag tatctaaacc atataacaag aactacgcaa
     1501 tgggagcgcc caacacgacc ggcatccgaa tattctagcc ctggcagacc tcttagctgc
     1561 tttgttgatg agaacactcc aattagtgga acaaatggtg caacatgtgg acagtcttca
     1621 gatcccaggc tggcagagag gagagtcagg tcacaacgac atagaaatta catgagcaga
     1681 acacatttac atactcctcc agacctacca gaaggctatg aacagaggac aacgcaacaa
     1741 ggccaggtgt atttcttaca tacacagact ggtgtgagca catggcatga tccaagagtg
     1801 cccagggatc ttagcaacat caattgtgaa gagcttggtc cattgcctcc tggatgggag
     1861 atccgtaata cggcaacagg cagagtttat ttcgttgacc ataacaacag aacaacacaa
     1921 tttacggatc ctcggctgtc tgctaacttg catttagttt taaatcggca gaaccaattg
     1981 aaagaccaac agcaacagca agtggtatcg ttatgtcctg atgacacaga atgcctgaca
     2041 gtcccaaggt acaagcgaga cctggttcag aaactaaaaa ttttgcggca agaactttcc
     2101 caacaacagc ctcaggcagg tcattgccgc attgaggttt ccagggaaga gatttttgag
     2161 gaatcatatc gacaggtcat gaaaatgaga ccaaaagatc tctggaagcg attaatgata
     2221 aaatttcgtg gagaagaagg ccttgactat ggaggcgttg ccagggaatg gttgtatctc
     2281 ttgtcacatg aaatgttgaa tccatactat ggcctcttcc agtattcaag agatgatatt
     2341 tatacattgc agatcaatcc tgattctgca gttaatccgg aacatttatc ctatttccac
     2401 tttgttggac gaataatggg aatggctgtg tttcatggac attatattga tggtggtttc
     2461 acattgcctt tttataagca attgcttggg aagtcaatta ccttggatga catggagtta
     2521 gtagatccgg atcttcacaa cagtttagtg tggatacttg agaatgatat tacaggtgtt
     2581 ttggaccata ccttctgtgt tgaacataat gcatatggtg aaattattca gcatgaactt
     2641 aaaccaaatg gcaaaagtat ccctgttaat gaagaaaata aaaaagaata tgtcaggctc
     2701 tatgtgaact ggagattttt acgaggcatt gaggctcaat tcttggctct gcagaaagga
     2761 tttaatgaag taattccaca acatctgctg aagacatttg atgagaagga gttagagctc
     2821 attatttgtg gacttggaaa gatagatgtt aatgactgga aggtaaacac ccggttaaaa
     2881 cactgtacac cagacagcaa cattgtcaaa tggttctgga aagctgtgga gttttttgat
     2941 gaagagcgac gagcaagatt gcttcagttt gtgacaggat cctctcgagt gcctctgcag
     3001 ggcttcaaag cattgcaagg tgctgcaggc ccgagactct ttaccataca ccagattgat
     3061 gcctgcacta acaacctgcc gaaagcccac acttgcttca atcgaataga cattccaccc
     3121 tatgaaagct atgaaaagct atatgaaaag ctgctaacag ccattgaaga aacatgtgga
     3181 tttgctgtgg aatgacaatc tagacccggg tggcatccct gtgacccctc cccagtgcct
     3241 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
     3301 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
     3361 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
     3421 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
     3481 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
     3541 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
     3601 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
     3661 accactgctc ccttccctgt ccttctgatt ttaaaataac tataccagca ggaggacgtc
     3721 cagacacagc ataggctacc tggccatgcc caaccggtgg gacatttgag ttgcttgctt
     3781 ggcactgtcc tctcatgcgt tgggtccact cagtagatgc ctgttgaatt gggtacgcgg
     3841 ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3901 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3961 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4021 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4081 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4141 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcctcgag
     4201 gaactgaaaa accagaaagt taattcccta tagtgagtcg tattaaattc gtaatcatgt
     4261 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4321 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4381 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4441 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4501 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4561 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4621 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4681 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4741 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4801 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4861 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4921 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4981 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5041 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5101 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5161 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5221 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5281 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5341 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5401 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5461 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5521 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5581 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5641 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5701 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5761 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5821 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5881 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5941 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6001 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6061 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6121 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6181 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6241 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6301 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6361 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct
     6421 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     6481 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     6541 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     6601 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     6661 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     6721 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     6781 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     6841 ttaacaaaat attaacgctt acaatttcca ttcgccattc aggctgcgca actgttggga
     6901 a
//
