LOCUS       sequence-349335-        6727 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 6727)
  AUTHORS   Gao S, Alarcon C, Sapkota G, Rahman S, Chen PY, Goerner N, Macias 
            MJ, Erdjument-Bromage H, Tempst P, Massague J
  TITLE     Ubiquitin ligase Nedd4L targets activated Smad2/3 to limit TGF-beta 
            signaling.
  JOURNAL   Mol Cell. 2009 Nov 13. 36(3):457-68.
  PUBMED    19917253
REFERENCE   2  (bases 1 to 6727)
  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. 
            2009 Nov 13. 36(3):457-68."
COMMENT     Sequence Label: pCI HA NEDD4 DD
FEATURES             Location/Qualifiers
     source          1..6727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        42..421
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        422..625
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     576..596
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     622..646
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          761..893
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     876..895
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        938..956
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             977..1003
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3745..3866
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3756..3775)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3810..3829
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4047..4502
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4134..4153)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4344..4365
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4569..4588)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4688..4710
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4748..4766)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4834..4938
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4939..5007,5008..5799)
                     /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(5157..5176)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5970..6558
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6459..6478
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtacatttat attggctcat gtccaatatg accgccatgt tggcattgat tattgactag
       61 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      121 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      181 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      241 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      301 tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      361 gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      421 ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt
      481 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      541 ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg
      601 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcacta gaagctttat
      661 tgcggtagtt tatcacagtt aaattgctaa cgcagtcagt gcttctgaca caacagtctc
      721 gaacttaagc tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag
      781 acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt
      841 ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca
      901 ctcccagttc aattacagct cttaaggcta gagtacttaa tacgactcac tataggctag
      961 cctcgagacc atggattatc cgtatgatgt tcctgattat gcagaattca cgcgtggtac
     1021 cgcaacttgc gcggtggagg tgttcgggct cctggaggac gaggaaaatt cacgaattgt
     1081 gagagtaaga gttatagccg gaataggcct tgccaagaag gatatattgg gagctagtga
     1141 tccttacgtg agagtgacgt tatatgaccc aatgaatgga gttcttacaa gtgtgcaaac
     1201 aaaaaccatt aaaaagagtt tgaatccaaa gtggaatgaa gaaatattat tcagagttca
     1261 tcctcagcag caccggcttc tttttgaagt gtttgacgaa aaccgattga caagagatga
     1321 tttcctaggt caagtggatg ttccacttta tccattaccg acagaaaatc caagattgga
     1381 gagaccatat acatttaagg attttgttct tcatccaaga agtcacaaat caagagttaa
     1441 aggttatctg agactaaaaa tgacttattt acctaaaacc agtggctcag aagatgataa
     1501 tgcagaacag gctgaggaat tagagcctgg ctgggttgtt ttggaccaac cagatgctgc
     1561 ttgccatttg cagcaacaac aagaaccttc tcctctacct ccagggtggg aagagaggca
     1621 ggatatcctt ggaaggacct attatgtaaa ccatgaatct agaagaacac agtggaaaag
     1681 accaacccct caggacaacc taacagatgc tgagaatggc aacattcaac tgcaagcaca
     1741 acgtgcattt accaccaggc ggcagatatc cgaggaaaca gaaagtgttg acaaccaaga
     1801 gtcttccgag aactgggaaa ttataagaga agatgaagcc accatgtata gcagccaggc
     1861 cttcccatca cctccaccgt caagtaactt ggatgttcca actcatcttg cagaagaatt
     1921 gaatgccaga ctcaccattt ttggaaattc agccgtgagc cagccagcat cgagctcaaa
     1981 tcattccagc agaagaggca gcttacaagc ctatactttt gaggaacaac ctacacttcc
     2041 tgtgcttttg cctacttcat ctggattacc accaggttgg gaagaaaaac aagatgaaag
     2101 aggaagatca tattatgtag atcacaattc cagaacgact acttggacaa agcccactgt
     2161 acaggccaca gtggagacca gtcagctgac ctcaagccag agttctgcag gccctcaatc
     2221 acaagcctcc accagtgatt caggccagca ggtgacccag ccatctgaaa ttgagcaagg
     2281 attccttcct aaaggctggg aagtccggca tgcaccaaat gggaggcctt tctttattga
     2341 ccacaacact aaaaccacca cctgggaaga tccaagattg aaaattccag cccatctgag
     2401 aggaaagaca tcacttgata cttccaatga tctagggcct ttacctccag gatgggaaga
     2461 gagaactcac acagatggaa gaatcttcta cataaatcac aatataaaaa gaacacaatg
     2521 ggaagatcct cggttggaga atgtagcaat aactggacca gcagtgccct actccaggga
     2581 ttacaaaaga aagtatgagt tcttccgaag aaagttgaag aagcagaatg acattccaaa
     2641 caaatttgaa atgaaacttc gccgagcaac tgttcttgaa gactcttacc ggagaattat
     2701 gggtgtcaag agagcagact tcctgaaggc tcgactgtgg attgagtttg atggtgaaaa
     2761 gggattggat tatggaggag ttgccagaga atggttcttc ctgatctcaa aggaaatgtt
     2821 taacccttat tatgggttgt ttgaatattc tgctacggac aattataccc tacagataaa
     2881 tccaaactct ggattgtgta acgaagatca cctctcttac ttcaagttta ttggtcgggt
     2941 agctggaatg gcagtttatc atggcaaact gttggatggt tttttcatcc gcccatttta
     3001 caagatgatg cttcacaaac caataaccct tcatgatatg gaatctgtgg atagtgaata
     3061 ttacaattcc ctaagatgga ttcttgaaaa tgacccaaca gaattggacc tcaggtttat
     3121 catagatgaa gaactttttg gacagacaca tcaacatgag ctgaaaaatg gtggatcaga
     3181 aatagttgtc accaataaga acaaaaagga atatatttat cttgtaatac aatggcgatt
     3241 tgtaaaccga atccagaagc aaatggctgc ttttaaagag ggattctttg aactaatacc
     3301 acaggatctc atcaaaattt ttgatgaaaa tgaactagag cttcttatgt gtggactggg
     3361 agatgttgat gtgaatgact ggagggaaca tacaaagtat aaaaatggct acagtgcaaa
     3421 tcatcaggtt atacagtggt tttggaaggc tgttttaatg atggattcag aaaaaagaat
     3481 aagattactt cagtttgtca ctggcacatc tcgggtgcct atgaatggat ttgctgaact
     3541 atacggttca aatggaccac agtcatttac agttgaacag tggggtacgc ctgaaaagct
     3601 gccaagagct cataccgctt ttaatcgcct ggacttgcca ccttatgaat catttgaaga
     3661 attatgggat aaacttcaga tggcaattga aaacacccag ggctttgatg gagttgatta
     3721 ggcggccgct tcgagcagac atgataagat acattgatga gtttggacaa accacaacta
     3781 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     3841 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     3901 ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa
     3961 tcgataagga tccgggctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     4021 agttgcgcag cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg
     4081 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4141 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4201 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4261 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4321 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4381 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4441 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     4501 ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     4561 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     4621 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4681 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     4741 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4801 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4861 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4921 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4981 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5041 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5101 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5161 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5221 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5281 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5341 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5401 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5461 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5521 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5581 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5641 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5701 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5761 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5821 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5881 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5941 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6001 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6061 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6121 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6181 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6241 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6301 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6361 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6421 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6481 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6541 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6601 tggccttttg ctcacatggc tcgacagatc ttcaatattg gccattagcc atattattca
     6661 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
     6721 ataatat
//
