LOCUS       sequence-372909-        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
FEATURES             Location/Qualifiers
     source          1..6727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(280..868)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(360..379)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1039..1830,1831..1899))
                     /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     1662..1681
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1900..2004)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2072..2090
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2128..2150)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2250..2269
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2336..2791)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2473..2494)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2685..2704
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2972..3093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3009..3028)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3063..3082
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5835..5861)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(5882..5900)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5943..5962)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          5945..6077
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     complement(6192..6216)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(6213..6416)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6242..6262)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(6417..6727,1..69)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata
       61 atcaatgcca acatggcggt catattggac atgagccaat ataaatgtac atattatgat
      121 atagatacaa cgtatgcaat ggccaatagc caatattgat ttatgctata taaccaatga
      181 ataatatggc taatggccaa tattgaagat ctgtcgagcc atgtgagcaa aaggccagca
      241 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
      301 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
      361 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
      421 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
      481 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
      541 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
      601 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
      661 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
      721 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
      781 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
      841 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
      901 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
      961 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     1021 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     1081 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     1141 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     1201 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     1261 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     1321 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     1381 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     1441 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     1501 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     1561 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     1621 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     1681 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     1741 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     1801 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     1861 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     1921 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     1981 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     2041 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     2101 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     2161 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     2221 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     2281 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
     2341 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2401 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2461 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2521 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     2581 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2641 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2701 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     2761 gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt
     2821 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gcccggatcc ttatcgattt
     2881 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacatctcc ccctgaacct
     2941 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     3001 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3061 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgctcgaagc ggccgcctaa
     3121 tcaactccat caaagccctg ggtgttttca attgccatct gaagtttatc ccataattct
     3181 tcaaatgatt cataaggtgg caagtccagg cgattaaaac aggtatgagc tcttggcagc
     3241 ttttcaggcg taccccactg ttcaactgta aatgactgtg gtccatttga accgtatagt
     3301 tcagcaaatc cattcatagg cacccgagat gtgccagtga caaactgaag taatcttatt
     3361 cttttttctg aatccatcat taaaacagcc ttccaaaacc actgtataac ctgatgattt
     3421 gcactgtagc catttttata ctttgtatgt tccctccagt cattcacatc aacatctccc
     3481 agtccacaca taagaagctc tagttcattt tcatcaaaaa ttttgatgag atcctgtggt
     3541 attagttcaa agaatccctc tttaaaagca gccatttgct tctggattcg gtttacaaat
     3601 cgccattgta ttacaagata aatatattcc tttttgttct tattggtgac aactatttct
     3661 gatccaccat ttttcagctc atgttgatgt gtctgtccaa aaagttcttc atctatgata
     3721 aacctgaggt ccaattctgt tgggtcattt tcaagaatcc atcttaggga attgtaatat
     3781 tcactatcca cagattccat atcatgaagg gttattggtt tgtgaagcat catcttgtaa
     3841 aatgggcgga tgaaaaaacc atccaacagt ttgccatgat aaactgccat tccagctacc
     3901 cgaccaataa acttgaagta agagaggtga tcttcgttac acaatccaga gtttggattt
     3961 atctgtaggg tataattgtc cgtagcagaa tattcaaaca acccataata agggttaaac
     4021 atttcctttg agatcaggaa gaaccattct ctggcaactc ctccataatc caatcccttt
     4081 tcaccatcaa actcaatcca cagtcgagcc ttcaggaagt ctgctctctt gacacccata
     4141 attctccggt aagagtcttc aagaacagtt gctcggcgaa gtttcatttc aaatttgttt
     4201 ggaatgtcat tctgcttctt caactttctt cggaagaact catactttct tttgtaatcc
     4261 ctggagtagg gcactgctgg tccagttatt gctacattct ccaaccgagg atcttcccat
     4321 tgtgttcttt ttatattgtg atttatgtag aagattcttc catctgtgtg agttctctct
     4381 tcccatcctg gaggtaaagg ccctagatca ttggaagtat caagtgatgt ctttcctctc
     4441 agatgggctg gaattttcaa tcttggatct tcccaggtgg tggttttagt gttgtggtca
     4501 ataaagaaag gcctcccatt tggtgcatgc cggacttccc agcctttagg aaggaatcct
     4561 tgctcaattt cagatggctg ggtcacctgc tggcctgaat cactggtgga ggcttgtgat
     4621 tgagggcctg cagaactctg gcttgaggtc agctgactgg tctccactgt ggcctgtaca
     4681 gtgggctttg tccaagtagt cgttctggaa ttgtgatcta cataatatga tcttcctctt
     4741 tcatcttgtt tttcttccca acctggtggt aatccagatg aagtaggcaa aagcacagga
     4801 agtgtaggtt gttcctcaaa agtataggct tgtaagctgc ctcttctgct ggaatgattt
     4861 gagctcgatg ctggctggct cacggctgaa tttccaaaaa tggtgagtct ggcattcaat
     4921 tcttctgcaa gatgagttgg aacatccaag ttacttgacg gtggaggtga tgggaaggcc
     4981 tggctgctat acatggtggc ttcatcttct cttataattt cccagttctc ggaagactct
     5041 tggttgtcaa cactttctgt ttcctcggat atctgccgcc tggtggtaaa tgcacgttgt
     5101 gcttgcagtt gaatgttgcc attctcagca tctgttaggt tgtcctgagg ggttggtctt
     5161 ttccactgtg ttcttctaga ttcatggttt acataatagg tccttccaag gatatcctgc
     5221 ctctcttccc accctggagg tagaggagaa ggttcttgtt gttgctgcaa atggcaagca
     5281 gcatctggtt ggtccaaaac aacccagcca ggctctaatt cctcagcctg ttctgcatta
     5341 tcatcttctg agccactggt tttaggtaaa taagtcattt ttagtctcag ataaccttta
     5401 actcttgatt tgtgacttct tggatgaaga acaaaatcct taaatgtata tggtctctcc
     5461 aatcttggat tttctgtcgg taatggataa agtggaacat ccacttgacc taggaaatca
     5521 tctcttgtca atcggttttc gtcaaacact tcaaaaagaa gccggtgctg ctgaggatga
     5581 actctgaata atatttcttc attccacttt ggattcaaac tctttttaat ggtttttgtt
     5641 tgcacacttg taagaactcc attcattggg tcatataacg tcactctcac gtaaggatca
     5701 ctagctccca atatatcctt cttggcaagg cctattccgg ctataactct tactctcaca
     5761 attcgtgaat tttcctcgtc ctccaggagc ccgaacacct ccaccgcgca agttgcggta
     5821 ccacgcgtga attctgcata atcaggaaca tcatacggat aatccatggt ctcgaggcta
     5881 gcctatagtg agtcgtatta agtactctag ccttaagagc tgtaattgaa ctgggagtgg
     5941 acacctgtgg agagaaaggc aaagtggatg tcagtaagac caataggtgc ctatcagaaa
     6001 cgcaagagtc ttctctgtct cgacaagccc agtttctatt ggtctcctta aacctgtctt
     6061 gtaaccttga tacttacctg cccagtgcct cacgaccaac ttctgcagct taagttcgag
     6121 actgttgtgt cagaagcact gactgcgtta gcaatttaac tgtgataaac taccgcaata
     6181 aagcttctag tgatctgacg gttcactaaa cgagctctgc ttatatagac ctcccaccgt
     6241 acacgcctac cgcccatttg cgtcaacggg gcggggttat tacgacattt tggaaagtcc
     6301 cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat
     6361 ccccgtgagt caaaccgcta tccacgccca ttggtgtact gccaaaaccg catcaccatg
     6421 gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtcccg taaggtcatg
     6481 tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcggactt
     6541 ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat
     6601 tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc
     6661 aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg
     6721 cggaact
//
