LOCUS       sequence-127220-        6387 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 6387)
  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 6387)
  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-TFGbeta receptor II K277R
FEATURES             Location/Qualifiers
     source          1..6387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      148..283
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(335..354)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(336..354)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(368..384)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(368..384)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(381..403)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    392..408
                     /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(416..422,423..440,441..446))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    461..482
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(599..616)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(770..1358)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(850..869)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1529..2320,2321..2389))
                     /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     2152..2171
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2390..2494)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2521..2976
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2608..2627)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2818..2839
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3102..3124
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3116..3133
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3117..3133
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3294..3673
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3674..3877
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3828..3848
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5212..5223
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             5658..5675
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5704..6326
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5903..5922)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(6350..6370)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6355..6387,1..297)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
       61 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
      121 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
      181 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
      241 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
      301 ctcctcgagg aactgaaaaa ccagaaagtt aattccctat agtgagtcgt attaaattcg
      361 taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
      421 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
      481 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
      541 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
      601 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
      661 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      721 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
      781 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
      841 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
      901 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
      961 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1021 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1081 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1141 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1201 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1261 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1321 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1381 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1441 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     1501 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     1561 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1621 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     1681 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1741 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     1801 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     1861 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     1921 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     1981 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2041 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     2101 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     2161 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     2221 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2281 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2341 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     2401 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2461 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     2521 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2581 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2641 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2701 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2761 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2821 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     2881 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     2941 acgcgaattt taacaaaata ttaacgctta caatttccat tcgccattca ggctgcgcaa
     3001 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     3061 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     3121 aacgacggcc agtgccaagc tgatctatac attgaatcaa tattggcaat tagccatatt
     3181 agtcattggt tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct
     3241 atatcataat atgtacattt atattggctc atgtccaata tgaccgccat gttgacattg
     3301 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     3361 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     3421 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     3481 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     3541 tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     3601 tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat
     3661 cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga
     3721 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     3781 aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg
     3841 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagaattc
     3901 agatctggta ccgggccccc cctcgagtcg gtctatgacg agcagcgggg tctgccatgg
     3961 gtcgggggct gctcaggggc ctgtggccgc tgcacatcgt cctgtggacg cgtatcgcca
     4021 gcacgatccc accgcacgtt cagaagtcgg ttaataacga catgatagtc actgacaaca
     4081 acggtgcagt caagtttcca caactgtgta aattttgtga tgtgagattt tccacctgtg
     4141 acaaccagaa atcctgcatg agcaactgca gcatcacctc catctgtgag aagccacagg
     4201 aagtctgtgt ggctgtatgg agaaagaatg acgagaacat aacactagag acagtttgcc
     4261 atgaccccaa gctcccctac catgacttta ttctggaaga tgctgcttct ccaaagtgca
     4321 ttatgaagga aaaaaaaaag cctggtgaga ctttcttcat gtgttcctgt agctctgatg
     4381 agtgcaatga caacatcatc ttctcagaag aatataacac cagcaatcct gacttgttgc
     4441 tagtcatatt tcaagtgaca ggcatcagcc tcctgccacc actgggagtt gccatatctg
     4501 tcatcatcat cttctactgc taccgcgtta accggcagca gaagctgagt tcaacctggg
     4561 aaaccggcaa gacgcggaag ctcatggagt tcagcgagca ctgtgccatc atcctggaag
     4621 atgaccgctc tgacatcagc tccacgtgtg ccaacaacat caaccacaac acagagctgc
     4681 tgcccattga gctggacacc ctggtgggga aaggtcgctt tgctgaggtc tataaggcca
     4741 agctgaagca gaacacttca gagcagtttg agacagtggc agtccggatc tttccctatg
     4801 aggagtatgc ctcttggaag acagagaagg acatcttctc agacatcaat ctgaagcatg
     4861 agaacatact ccagttcctg acggctgagg agcggaagac ggagttgggg aaacaatact
     4921 ggctgatcac cgccttccac gccaagggca acctacagga gtacctgacg cggcatgtca
     4981 tcagctggga ggacctgcgc aagctgggca gctccctcgc ccgggggatt gctcacctcc
     5041 acagtgatca cactccatgt gggaggccca agatgcccat cgtgcacagg gacctcaaga
     5101 gctccaatat cctcgtgaag aacgacctaa cctgctgcct gtgtgacttt gggctttccc
     5161 tgcgtctgga ccctactctg tctgtggatg acctggctaa cagtgggcag gtgggaactg
     5221 caagatacat ggctccagaa gtcctagaat ccaggatgaa tttggagaat gttgagtcct
     5281 tcaagcagac cgatgtctac tccatggctc tggtgctctg ggaaatgaca tctcgctgta
     5341 atgcagtggg agaagtaaaa gattatgagc ctccatttgg ttccaaggtg cgggagcacc
     5401 cctgtgtcga aagcatgaag gacaacgtgt tgagagatcg agggcgacca gaaattccca
     5461 gcttctggct caaccaccag ggcatccaga tggtgtgtga gacgttgact gagtgctggg
     5521 accacgaccc agaggcccgt ctcacagccc agtgtgtggc agaacgcttc agtgagctgg
     5581 agcatctgga caggctctcg gggaggagct gctcggagga gaagattcct gaagacggct
     5641 ccctaaacac taccaaacat caccaccatc atcactagga attcctgcag cccgggggat
     5701 cccgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact
     5761 ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg
     5821 tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag
     5881 acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct
     5941 tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag
     6001 ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga
     6061 cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca
     6121 ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt
     6181 ctgattttaa aataactata ccagcaggag gacgtccaga cacagcatag gctacctggc
     6241 catgcccaac cggtgggaca tttgagttgc ttgcttggca ctgtcctctc atgcgttggg
     6301 tccactcagt agatgcctgt tgaattgggt acgcggccag cttggctgtg gaatgtgtgt
     6361 cagttagggt gtggaaagtc cccaggc
//
