LOCUS       sequence-177678-        7015 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 7015)
  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 7015)
  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 NEDD4L
FEATURES             Location/Qualifiers
     source          1..7015
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        91..470
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        471..674
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     625..645
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     671..695
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          810..942
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     925..944
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        987..1005
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1026..1052
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4082..4203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4093..4112)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4147..4166
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4384..4839
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4471..4490)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4681..4702
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4906..4925)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5025..5047
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5085..5103)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5171..5275
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5276..5344,5345..6136)
                     /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(5494..5513)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6307..6895
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6796..6815
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata
       61 ttggctcatg tccaatatga ccgccatgtt ggcattgatt attgactagt tattaatagt
      121 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      181 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      241 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      301 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta
      361 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg
      421 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      481 tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      541 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      601 gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      661 atataagcag agctcgttta gtgaaccgtc agatcactag aagctttatt gcggtagttt
      721 atcacagtta aattgctaac gcagtcagtg cttctgacac aacagtctcg aacttaagct
      781 gcagaagttg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag
      841 gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac
      901 ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccac tcccagttca
      961 attacagctc ttaaggctag agtacttaat acgactcact ataggctagc ctcgagacca
     1021 tggattatcc gtatgatgtt cctgattatg cagaattcac gcgtggtacc ccgggtgggt
     1081 ggctgcgcag ggccctacct gggcgggagc ggctgcagag ccctgtccac gcggtgcctc
     1141 cccagcacgg cacctcccac tccaggctgt tggttacctg gccgggagct ggcagagacc
     1201 aggatttctc ctcgccgccg ttgctgttgt taggggaaac agaccatctg catctagacc
     1261 tgcctctctc tccgctcccc acttcagacg agctcttcct gcctggaatc tgtgatccgt
     1321 atgtgaaact ttcattgtac gtagcggatg agaatagaga acttgctttg gtccagacaa
     1381 aaacaattaa aaagacactg aacccaaaat ggaatgaaga attttatttc agggtaaacc
     1441 catctaatca cagactccta tttgaagtat ttgacgaaaa tagactgaca cgagacgact
     1501 tcctgggcca ggtggacgtg ccccttagtc accttccgac agaagatcca accatggagc
     1561 gaccctatac atttaaggac tttctcctca gaccaagaag tcataagtct cgagttaagg
     1621 gatttttgcg attgaaaatg gcctatatgc caaaaaatgg aggtcaagat gaagaaaaca
     1681 gtgaccagag ggatgacatg gagcatggat gggaagttgt tgactcaaat gactcggctt
     1741 ctcagcacca agaggaactt cctcctcctc ctctgcctcc cgggtgggaa gaaaaagtgg
     1801 acaatttagg ccgaacttac tatgtcaacc acaacaaccg gaccactcag tggcacagac
     1861 caagcctgat ggacgtgtcc tcggagtcgg acaataacat cagacagatc aaccaggagg
     1921 cagcacaccg gcgcttccgc tcccgcaggc acatcagcga agacttggag cccgagccct
     1981 cggagggcgg ggatgtcccc gagccttggg agaccatttc agaggaagtg aatatcgctg
     2041 gagactctct cggtctggct ctgcccccac caccggcctc cccaggatct cggaccagcc
     2101 ctcaggagct gtcagaggaa ctaagcagaa ggcttcagat cactccagac tccaatgggg
     2161 aacagttcag ctctttgatt caaagagaac cctcctcaag gttgaggtca tgcagtgtca
     2221 ccgacgcagt tgcagaacag ggccatctac caccgccatc agtggcctat gtacatacca
     2281 cgccgggtct gccttcaggc tgggaagaaa gaaaagatgc taaggggcgc acatactatg
     2341 tcaatcataa caatcgaacc acaacttgga ctcgacctat catgcagctt gcagaagatg
     2401 gtgcgtccgg atcagccaca aacagtaaca accatctaat cgagcctcag atccgccggc
     2461 ctcgtagcct cagctcgcca acagtaactt tatctgcccc gctggagggt gccaaggact
     2521 cacccgtacg tcgggctgtg aaagacaccc tttccaaccc acagtcccca cagccatcac
     2581 cttacaactc ccccaaacca caacacaaag tcacacagag cttcttgcca cccggctggg
     2641 aaatgaggat agcgccaaac ggccggccct tcttcattga tcataacaca aagactacaa
     2701 cctgggaaga tccacgtttg aaatttccag tacatatgcg gtcaaagaca tctttaaacc
     2761 ccaatgacct tggccccctt cctcctggct gggaagaaag aattcacttg gatggccgaa
     2821 cgttttatat tgatcataat agcaaaatta ctcagtggga agacccaaga ctgcagaacc
     2881 cagctattac tggtccggct gtcccttact ccagagaatt taagcagaaa tatgactact
     2941 tcaggaagaa attaaagaaa cctgctgata tccccaatag gtttgaaatg aaacttcaca
     3001 gaaataacat atttgaagag tcctatcgga gaattatgtc cgtgaaaaga ccagatgtcc
     3061 taaaagctag actgtggatt gagtttgaat cagagaaagg tcttgactat gggggtgtgg
     3121 ccagagaatg gttcttctta ctgtccaaag agatgttcaa cccctactac ggcctctttg
     3181 agtactctgc cacggacaac tacacccttc agatcaaccc taattcaggc ctctgtaatg
     3241 aggatcattt gtcctacttc acttttattg gaagagttgc tggtctggcc gtatttcatg
     3301 ggaagctctt agatggtttc ttcattagac cattttacaa gatgatgttg ggaaagcaga
     3361 taaccctgaa tgacatggaa tctgtggata gtgaatatta caactctttg aaatggatcc
     3421 tggagaatga ccctactgag ctggacctca tgttctgcat agacgaagaa aactttggac
     3481 agacatatca agtggatttg aagcccaatg ggtcagaaat aatggtcaca aatgaaaaca
     3541 aaagggaata tatcgactta gtcatccagt ggagatttgt gaacagggtc cagaagcaga
     3601 tgaacgcctt cttggaggga ttcacagaac tacttcctat tgatttgatt aaaatttttg
     3661 acgaaaatga gctggagttg ctcatgtgcg gcctcggtga tgtggatgtg aatgactgga
     3721 gacagcattc tatttacaag aacggctact gcccaaacca ccccgtcatt cagtggttct
     3781 ggaaggctgt gctactcatg gacgccgaaa agcgtatccg gttactgcag tttgtcacag
     3841 ggacatcgcg agtacctatg aatggatttg ccgaacttta tggttccaat ggtcctcagc
     3901 tgtttacaat agagcaatgg ggcagtcctg agaaactgcc cagagctcac acatgcttta
     3961 atcgccttga cttacctcca tatgaaacct ttgaagattt acgagagaaa cttctcatgg
     4021 ccgtggaaaa tgctcaagga tttgaagggg tggattaagc ggccgcttcg agcagacatg
     4081 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     4141 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     4201 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     4261 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gggctggcgt
     4321 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     4381 tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4441 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4501 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4561 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4621 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4681 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4741 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4801 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct
     4861 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     4921 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     4981 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     5041 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     5101 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     5161 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5221 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5281 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5341 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5401 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5461 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5521 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5581 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5641 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5701 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5761 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5821 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5881 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5941 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6001 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6061 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6121 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6181 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6241 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6301 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6361 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6421 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6481 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6541 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6601 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6661 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6721 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6781 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6841 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6901 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatggctcg
     6961 acagatcttc aatattggcc attagccata ttattcattg gttatatagc ataaa
//
