LOCUS       40924_10751        6727 bp DNA     circular SYN 13-MAY-2021
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)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6727)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell. 
            2009 Nov 13. 36(3):457-68."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        139..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     primer_bind     718..742
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        1034..1052
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1073..1099
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     polyA_signal    complement(3841..3962)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      4143..4598
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4665..4684)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     4784..4806
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4844..4862)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        4930..5034
                     /label=AmpR promoter
     CDS             5035..5892
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6066..6654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tcgtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt acttaatacg actcactata ggctagcctc gagaccatgg attatccgta
     1081 tgatgttcct gattatgcag aattcacgcg tggtaccgca acttgcgcgg tggaggtgtt
     1141 cgggctcctg gaggacgagg aaaattcacg aattgtgaga gtaagagtta tagccggaat
     1201 aggccttgcc aagaaggata tattgggagc tagtgatcct tacgtgagag tgacgttata
     1261 tgacccaatg aatggagttc ttacaagtgt gcaaacaaaa accattaaaa agagtttgaa
     1321 tccaaagtgg aatgaagaaa tattattcag agttcatcct cagcagcacc ggcttctttt
     1381 tgaagtgttt gacgaaaacc gattgacaag agatgatttc ctaggtcaag tggatgttcc
     1441 actttatcca ttaccgacag aaaatccaag attggagaga ccatatacat ttaaggattt
     1501 tgttcttcat ccaagaagtc acaaatcaag agttaaaggt tatctgagac taaaaatgac
     1561 ttatttacct aaaaccagtg gctcagaaga tgataatgca gaacaggctg aggaattaga
     1621 gcctggctgg gttgttttgg accaaccaga tgctgcttgc catttgcagc aacaacaaga
     1681 accttctcct ctacctccag ggtgggaaga gaggcaggat atccttggaa ggacctatta
     1741 tgtaaaccat gaatctagaa gaacacagtg gaaaagacca acccctcagg acaacctaac
     1801 agatgctgag aatggcaaca ttcaactgca agcacaacgt gcatttacca ccaggcggca
     1861 gatatccgag gaaacagaaa gtgttgacaa ccaagagtct tccgagaact gggaaattat
     1921 aagagaagat gaagccacca tgtatagcag ccaggccttc ccatcacctc caccgtcaag
     1981 taacttggat gttccaactc atcttgcaga agaattgaat gccagactca ccatttttgg
     2041 aaattcagcc gtgagccagc cagcatcgag ctcaaatcat tccagcagaa gaggcagctt
     2101 acaagcctat acttttgagg aacaacctac acttcctgtg cttttgccta cttcatctgg
     2161 attaccacca ggttgggaag aaaaacaaga tgaaagagga agatcatatt atgtagatca
     2221 caattccaga acgactactt ggacaaagcc cactgtacag gccacagtgg agaccagtca
     2281 gctgacctca agccagagtt ctgcaggccc tcaatcacaa gcctccacca gtgattcagg
     2341 ccagcaggtg acccagccat ctgaaattga gcaaggattc cttcctaaag gctgggaagt
     2401 ccggcatgca ccaaatggga ggcctttctt tattgaccac aacactaaaa ccaccacctg
     2461 ggaagatcca agattgaaaa ttccagccca tctgagagga aagacatcac ttgatacttc
     2521 caatgatcta gggcctttac ctccaggatg ggaagagaga actcacacag atggaagaat
     2581 cttctacata aatcacaata taaaaagaac acaatgggaa gatcctcggt tggagaatgt
     2641 agcaataact ggaccagcag tgccctactc cagggattac aaaagaaagt atgagttctt
     2701 ccgaagaaag ttgaagaagc agaatgacat tccaaacaaa tttgaaatga aacttcgccg
     2761 agcaactgtt cttgaagact cttaccggag aattatgggt gtcaagagag cagacttcct
     2821 gaaggctcga ctgtggattg agtttgatgg tgaaaaggga ttggattatg gaggagttgc
     2881 cagagaatgg ttcttcctga tctcaaagga aatgtttaac ccttattatg ggttgtttga
     2941 atattctgct acggacaatt ataccctaca gataaatcca aactctggat tgtgtaacga
     3001 agatcacctc tcttacttca agtttattgg tcgggtagct ggaatggcag tttatcatgg
     3061 caaactgttg gatggttttt tcatccgccc attttacaag atgatgcttc acaaaccaat
     3121 aacccttcat gatatggaat ctgtggatag tgaatattac aattccctaa gatggattct
     3181 tgaaaatgac ccaacagaat tggacctcag gtttatcata gatgaagaac tttttggaca
     3241 gacacatcaa catgagctga aaaatggtgg atcagaaata gttgtcacca ataagaacaa
     3301 aaaggaatat atttatcttg taatacaatg gcgatttgta aaccgaatcc agaagcaaat
     3361 ggctgctttt aaagagggat tctttgaact aataccacag gatctcatca aaatttttga
     3421 tgaaaatgaa ctagagcttc ttatgtgtgg actgggagat gttgatgtga atgactggag
     3481 ggaacataca aagtataaaa atggctacag tgcaaatcat caggttatac agtggttttg
     3541 gaaggctgtt ttaatgatgg attcagaaaa aagaataaga ttacttcagt ttgtcactgg
     3601 cacatctcgg gtgcctatga atggatttgc tgaactatac ggttcaaatg gaccacagtc
     3661 atttacagtt gaacagtggg gtacgcctga aaagctgcca agagctcata cctgttttaa
     3721 tcgcctggac ttgccacctt atgaatcatt tgaagaatta tgggataaac ttcagatggc
     3781 aattgaaaac acccagggct ttgatggagt tgattaggcg gccgcttcga gcagacatga
     3841 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     3901 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     3961 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     4021 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg ggctggcgta
     4081 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     4141 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4201 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4261 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4321 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4381 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4441 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4501 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4561 taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt
     4621 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
     4681 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4741 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4801 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     4861 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4921 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4981 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5041 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5101 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5161 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5221 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5281 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5341 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5401 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5461 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5521 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5581 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5641 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5701 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5761 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5821 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5881 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5941 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6001 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6061 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6121 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6181 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6241 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6301 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6361 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6421 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6481 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6541 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6601 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6661 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catggctcga
     6721 cagatct
//
