LOCUS       sequence-220062-        6396 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 6396)
  AUTHORS   Wrana JL, Attisano L, Carcamo J, Zentella A, Doody J, Laiho M, Wang 
            XF, Massague J
  TITLE     TGF beta signals through a heteromeric protein kinase receptor 
            complex.
  JOURNAL   Cell. 1992 Dec 11. 71(6):1003-14.
  PUBMED    1333888
REFERENCE   2  (bases 1 to 6396)
  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: "Cell. 1992 
            Dec 11. 71(6):1003-14."
COMMENT     Sequence Label: pCMV5B-TGFbeta receptor II wt
FEATURES             Location/Qualifiers
     source          1..6396
                     /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"
     CDS             1981..1992
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             2427..2453
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     polyA_signal    2482..3104
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2681..2700)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3128..3148)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3293..3427
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3479..3498)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3480..3498)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3512..3528)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3512..3528)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3525..3547)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3536..3552
                     /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(3560..3566,3567..3584,3585..3590))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3605..3626
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3743..3760)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3914..4502)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3994..4013)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4673..5464,5465..5533))
                     /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     5296..5315
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5534..5638)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5665..6120
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5752..5771)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5962..5983
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6246..6268
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6260..6277
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6261..6277
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gtacatttat attggctcat gtccaatatg accgccatgt tgacattgat 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 cagaattcag atctggtacc
      661 acgcgtatcg ataagcttgc atgcctgcag gtcgagtcgg tctatgacga gcagcggggt
      721 ctgccatggg tcgggggctg ctcaggggcc tgtggccgct gcacatcgtc ctgtggacgc
      781 gtatcgccag cacgatccca ccgcacgttc agaagtcggt taataacgac atgatagtca
      841 ctgacaacaa cggtgcagtc aagtttccac aactgtgtaa attttgtgat gtgagatttt
      901 ccacctgtga caaccagaaa tcctgcatga gcaactgcag catcacctcc atctgtgaga
      961 agccacagga agtctgtgtg gctgtatgga gaaagaatga cgagaacata acactagaga
     1021 cagtttgcca tgaccccaag ctcccctacc atgactttat tctggaagat gctgcttctc
     1081 caaagtgcat tatgaaggaa aaaaaaaagc ctggtgagac tttcttcatg tgttcctgta
     1141 gctctgatga gtgcaatgac aacatcatct tctcagaaga atataacacc agcaatcctg
     1201 acttgttgct agtcatattt caagtgacag gcatcagcct cctgccacca ctgggagttg
     1261 ccatatctgt catcatcatc ttctactgct accgcgttaa ccggcagcag aagctgagtt
     1321 caacctggga aaccggcaag acgcggaagc tcatggagtt cagcgagcac tgtgccatca
     1381 tcctggaaga tgaccgctct gacatcagct ccacgtgtgc caacaacatc aaccacaaca
     1441 cagagctgct gcccattgag ctggacaccc tggtggggaa aggtcgcttt gctgaggtct
     1501 ataaggccaa gctgaagcag aacacttcag agcagtttga gacagtggca gtcaagatct
     1561 ttccctatga ggagtatgcc tcttggaaga cagagaagga catcttctca gacatcaatc
     1621 tgaagcatga gaacatactc cagttcctga cggctgagga gcggaagacg gagttgggga
     1681 aacaatactg gctgatcacc gccttccacg ccaagggcaa cctacaggag tacctgacgc
     1741 ggcatgtcat cagctgggag gacctgcgca agctgggcag ctccctcgcc cgggggattg
     1801 ctcacctcca cagtgatcac actccatgtg ggaggcccaa gatgcccatc gtgcacaggg
     1861 acctcaagag ctccaatatc ctcgtgaaga acgacctaac ctgctgcctg tgtgactttg
     1921 ggctttccct gcgtctggac cctactctgt ctgtggatga cctggctaac agtgggcagg
     1981 tgggaactgc aagatacatg gctccagaag tcctagaatc caggatgaat ttggagaatg
     2041 ttgagtcctt caagcagacc gatgtctact ccatggctct ggtgctctgg gaaatgacat
     2101 ctcgctgtaa tgcagtggga gaagtaaaag attatgagcc tccatttggt tccaaggtgc
     2161 gggagcaccc ctgtgtcgaa agcatgaagg acaacgtgtt gagagatcga gggcgaccag
     2221 aaattcccag cttctggctc aaccaccagg gcatccagat ggtgtgtgag acgttgactg
     2281 agtgctggga ccacgaccca gaggcccgtc tcacagccca gtgtgtggca gaacgcttca
     2341 gtgagctgga gcatctggac aggctctcgg ggaggagctg ctcggaggag aagattcctg
     2401 aagacggctc cctaaacact accaaatatg atgttcctga ttatgctagc ctctaggaat
     2461 tcctgcagcc cgggggatcc cgggtggcat ccctgtgacc cctccccagt gcctctcctg
     2521 gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc
     2581 attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc
     2641 aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg
     2701 agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc
     2761 ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt
     2821 ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa
     2881 tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact
     2941 gctcccttcc ctgtccttct gattttaaaa taactatacc agcaggagga cgtccagaca
     3001 cagcataggc tacctggcca tgcccaaccg gtgggacatt tgagttgctt gcttggcact
     3061 gtcctctcat gcgttgggtc cactcagtag atgcctgttg aattgggtac gcggccagct
     3121 tggctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3181 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3241 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3301 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3361 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3421 ggaggcctag gcttttgcaa aaagctcctc gaggaactga aaaaccagaa agttaattcc
     3481 ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt
     3541 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     3601 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3661 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3721 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3781 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3841 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3901 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3961 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4021 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4081 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4141 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4201 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4261 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4321 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4381 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4441 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4501 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4561 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4621 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4681 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4741 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4801 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4861 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4921 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4981 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5041 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5101 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5161 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5221 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5281 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5341 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5401 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5461 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5521 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5581 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5641 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5701 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     5761 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5821 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5881 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5941 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     6001 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     6061 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     6121 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6181 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     6241 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
     6301 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
     6361 ttggccattg catacgttgt atctatatca taatat
//
