LOCUS       sequence-127174-        6256 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 6256)
  AUTHORS   Wieser R, Wrana JL, Massague J
  TITLE     GS domain mutations that constitutively activate T beta R-I, the 
            downstream signaling component in the TGF-beta receptor complex.
  JOURNAL   EMBO J. 1995 May 15. 14(10):2199-208.
  PUBMED    7774578
REFERENCE   2  (bases 1 to 6256)
  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: "EMBO J. 
            1995 May 15. 14(10):2199-208."
COMMENT     Sequence Label: pCMV5B-TGFbeta receptor I K232R
FEATURES             Location/Qualifiers
     source          1..6256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1933..2388
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2020..2039)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2230..2251
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2514..2536
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2528..2545
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2529..2545
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        2706..3085
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3086..3289
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3240..3260
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4951..4968
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4985..5607
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5184..5203)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(5631..5651)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5636..5965
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5816..5951
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6003..6022)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6004..6022)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6036..6052)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6036..6052)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6049..6071)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6060..6076
                     /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(6084..6090,6091..6108,6109..6114))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6129..6150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     1981 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2041 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2101 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2161 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2221 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2281 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2341 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     2521 acgacgttgt aaaacgacgg ccagtgccaa gctgatctat acattgaatc aatattggca
     2581 attagccata ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca
     2641 tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc
     2701 atgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     2761 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     2821 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     2881 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2941 acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc
     3001 cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta
     3061 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg
     3121 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     3181 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac
     3241 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
     3301 ccgtcagaat tcagatctgg taccacgcgt atcgataagc ttgatatcga attccgttgc
     3361 tgtcgggcga ggcgaggttt gctggggtga ggcagcggcg cggccgggcc gggccgggcc
     3421 acaggcggtg gcggcgggac catggaggcg gcggtcgctg ctccgcgtcc ccggctgctc
     3481 ctcctcgtgc tggcggcggc ggcggcggcg gcggcggcgc tgctcccggg ggcgacggcg
     3541 ttacagtgtt tctgccacct ctgtacaaaa gacaatttta cttgtgtgac agatgggctc
     3601 tgctttgtct ctgtcacaga gaccacagac aaagttatac acaacagcat gtgtatagct
     3661 gaaattgact taattcctcg agataggccg tttgtatgtg caccctcttc aaaaactggg
     3721 tctgtgacta caacatattg ctgcaatcag gaccattgca ataaaataga acttccaact
     3781 actgtaaagt catcacctgg ccttggtcct gtggaactgg cagctgtcat tgctggacca
     3841 gtgtgcttcg tctgcatctc actcatgttg atggtctata tctgccacaa ccgcactgtc
     3901 attcaccatc gagtgccaaa tgaagaggac ccttcattag atcgcccttt tatttcagag
     3961 ggtactacgt tgaaagactt aatttatgat atgacaacgt caggttctgg ctcaggttta
     4021 ccattgcttg ttcagagaac aattgcgaga actattgtgt tacaagaaag cattggcaaa
     4081 ggtcgatttg gagaagtttg gagaggaaag tggcggggag aagaagttgc tgttcggata
     4141 ttctcctcta gagaagaacg ttcgtggttc cgtgaggcag agatttatca aactgtaatg
     4201 ttacgtcatg aaaacatcct gggatttata gcagcagaca ataaagacaa tggtacttgg
     4261 actcagctct ggttggtgtc agattatcat gagcatggat ctctttttga ttacttaaac
     4321 agatacacag ttactgtgga aggaatgata aaacttgctc tgtccacggc gagcggtctt
     4381 gcccatcttc acatggagat tgttggtacc caaggaaagc cagccattgc tcatagagat
     4441 ttgaaatcaa agaatatctt ggtaaagaag aatggaactt gctgtattgc agacttagga
     4501 ctggcagtaa gacatgattc agccacagat accattgata ttgctccaaa ccacagagtg
     4561 ggaacaaaaa ggtacatggc ccctgaagtt ctcgatgatt ccataaatat gaaacatttt
     4621 gaatccttca aacgtgctga catctatgca atgggcttag tattctggga aattgctcga
     4681 cgatgttcca ttggtggaat tcatgaagat taccaactgc cttattatga tcttgtacct
     4741 tctgacccat cagttgaaga aatgagaaaa gttgtttgtg aacagaagtt aaggccaaat
     4801 atcccaaaca gatggcagag ctgtgaagcc ttgagagtaa tggctaaaat tatgagagaa
     4861 tgttggtatg ccaatggagc agctaggctt acagcattgc ggattaagaa aacattatcg
     4921 caactcagtc aacaggaagg catcaaaatg catcaccacc atcatcacta aactagtgga
     4981 tcccgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac
     5041 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt
     5101 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa
     5161 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc
     5221 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga
     5281 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag
     5341 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc
     5401 accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct
     5461 tctgatttta aaataactat accagcagga ggacgtccag acacagcata ggctacctgg
     5521 ccatgcccaa ccggtgggac atttgagttg cttgcttggc actgtcctct catgcgttgg
     5581 gtccactcag tagatgcctg ttgaattggg tacgcggcca gcttggctgt ggaatgtgtg
     5641 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5701 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5761 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5821 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5881 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     5941 ttttggaggc ctaggctttt gcaaaaagct cctcgaggaa ctgaaaaacc agaaagttaa
     6001 ttccctatag tgagtcgtat taaattcgta atcatgtcat agctgtttcc tgtgtgaaat
     6061 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6121 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6181 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6241 ttgcgtattg ggcgct
//
