LOCUS       sequence-364716-        6458 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 6458)
  TITLE     Joan Massague lab plasmids
REFERENCE   2  (bases 1 to 6458)
  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"
COMMENT     Sequence Label: pCMV5 HA-TGFBRII
FEATURES             Location/Qualifiers
     source          1..6458
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             763..789
                     /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"
     CDS             1967..1978
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    2483..3105
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2682..2701)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3129..3149)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3134..3463
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3314..3449
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3501..3520)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3502..3520)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3534..3550)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3534..3550)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3547..3569)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3558..3574
                     /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(3582..3588,3589..3606,3607..3612))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3627..3648
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3765..3782)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3936..4524)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4016..4035)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4695..5486,5487..5555))
                     /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     5318..5337
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5556..5660)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5687..6141
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5774..5793)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5984..6005
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6267..6289
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6281..6298
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6282..6298
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattcaga tctggtacca cgcgtatcga taagcttgca tgcctgcagg tcgagtcggt
      661 ctatgacgag cagcggggtc tgccatgggt cgggggctgc tcaggggcct gtggccgctg
      721 cacatcgtcc tgtggacgcg tatcgccagc acgatcccac cgtatgatgt tcctgattat
      781 gctagcctcc acgttcagaa gtcggttaat aacgacatga tagtcactga caacaacggt
      841 gcagtcaagt ttccacaact gtgtaaattt tgtgatgtga gattttccac ctgtgacaac
      901 cagaaatcct gcatgagcaa ctgcagcatc acctccatct gtgagaagcc acaggaagtc
      961 tgtgtggctg tatggagaaa gaatgacgag aacataacac tagagacagt ttgccatgac
     1021 cccaagctcc cctaccatga ctttattctg gaagatgctg cttctccaaa gtgcattatg
     1081 aaggaaaaaa aaaagcctgg tgagactttc ttcatgtgtt cctgtagctc tgatgagtgc
     1141 aatgacaaca tcatcttctc agaagaatat aacaccagca atcctgactt gttgctagtc
     1201 atatttcaag tgacaggcat cagcctcctg ccaccactgg gagttgccat atctgtcatc
     1261 atcatcttct actgctaccg cgttaaccgg cagcagaagc tgagttcaac ctgggaaacc
     1321 ggcaagacgc ggaagctcat ggagttcagc gagcactgtg ccatcatcct ggaagatgac
     1381 cgctctgaca tcagctccac gtgtgccaac aacatcaacc acaacacaga gctgctgccc
     1441 attgagctgg acaccctggt ggggaaaggt cgctttgctg aggtctataa ggccaagctg
     1501 aagcagaaca cttcagagca gtttgagaca gtggcagtca agatctttcc ctatgaggag
     1561 tatgcctctt ggaagacaga gaaggacatc ttctcagaca tcaatctgaa gcatgagaac
     1621 atactccagt tcctgacggc tgaggagcgg aagacggagt tggggaaaca atactggctg
     1681 atcaccgcct tccacgccaa gggcaaccta caggagtacc tgacgcggca tgtcatcagc
     1741 tgggaggacc tgcgcaagct gggcagctcc ctcgcccggg ggattgctca cctccacagt
     1801 gatcacactc catgtgggag gcccaagatg cccatcgtgc acagggacct caagagctcc
     1861 aatatcctcg tgaagaacga cctaacctgc tgcctgtgtg actttgggct ttccctgcgt
     1921 ctggacccta ctctgtctgt ggatgacctg gctaacagtg ggcaggtggg aactgcaaga
     1981 tacatggctc cagaagtcct agaatccagg atgaatttgg agaatgttga gtccttcaag
     2041 cagaccgatg tctactccat ggctctggtg ctctgggaaa tgacatctcg ctgtaatgca
     2101 gtgggagaag taaaagatta tgagcctcca tttggttcca aggtgcggga gcacccctgt
     2161 gtcgaaagca tgaaggacaa cgtgttgaga gatcgagggc gaccagaaat tcccagcttc
     2221 tggctcaacc accagggcat ccagatggtg tgtgagacgt tgactgagtg ctgggaccac
     2281 gacccagagg cccgtctcac agcccagtgt gtggcagaac gcttcagtga gctggagcat
     2341 ctggacaggc tctcggggag gagctgctcg gaggagaaga ttcctgaaga cggctcccta
     2401 aacactacca aatagctctt ctggggcagg ctgggccatg tccaaagagg ctgccccgaa
     2461 ttcctgcagc ccgggggatc ccgggtggca tccctgtgac ccctccccag tgcctctcct
     2521 ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat
     2581 cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag
     2641 caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg
     2701 gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct
     2761 cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt
     2821 tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta
     2881 atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac
     2941 tgctcccttc cctgtccttc tgattttaaa ataactatac cagcaggagg acgtccagac
     3001 acagcatagg ctacctggcc atgcccaacc ggtgggacat ttgagttgct tgcttggcac
     3061 tgtcctctca tgcgttgggt ccactcagta gatgcctgtt gaattgggta cgcggccagc
     3121 ttggctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3181 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3241 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3301 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3361 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3421 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc tcgaggaact
     3481 gaaaaaccag aaagttaatt ccctatagtg agtcgtatta aattcgtaat catgtcatag
     3541 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3601 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3661 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3721 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3781 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3841 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3901 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3961 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4021 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4081 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4141 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4201 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4261 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4321 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4381 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4441 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4501 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4561 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4621 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4681 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4741 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4801 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4861 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4921 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4981 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5041 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5101 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5161 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5221 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5281 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5341 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5401 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5461 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5521 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5581 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5641 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc
     5701 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     5761 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     5821 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     5881 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     5941 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     6001 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     6061 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaatttaaca
     6121 aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg
     6181 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     6241 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
     6301 caagctgatc tatacattga atcaatattg gcaattagcc atattagtca ttggttatat
     6361 agcataaatc aatattggct attggccatt gcatacgttg tatctatatc ataatatgta
     6421 catttatatt ggctcatgtc caatatgacc gccatgtt
//
