LOCUS       sequence-190161-        6181 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 6181)
  AUTHORS   Feng XH, Derynck R
  TITLE     Ligand-independent activation of transforming growth factor (TGF) 
            beta signaling pathways by heteromeric cytoplasmic domains of 
            TGF-beta receptors.
  JOURNAL   J Biol Chem. 1996 May 31. 271(22):13123-9.
  PUBMED    8662796
REFERENCE   2  (bases 1 to 6181)
  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: "J Biol 
            Chem. 1996 May 31. 271(22):13123-9."
COMMENT     Sequence Label: pRK5 TGF beta type I receptor (T202D) Flag
FEATURES             Location/Qualifiers
     source          1..6181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    232..253
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(268..273,274..291,292..298)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    306..322
                     /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)."
     primer_bind     311..333
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     330..346
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     330..346
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        375..754
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        755..958
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     909..929
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     955..979
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1189..1207
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1189..1206
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1220..1239
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             2807..2830
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      2842..2851
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2855..2989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2892..2911)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2946..2965
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3058..3415
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3058..3249
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3081..3101)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3266..3401
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3435..3452)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3435..3451)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3444..3466)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3664..4119
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3751..3770)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3961..3982
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4136..4155)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4255..4277
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4315..4333)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4401..4505
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4506..4574,4575..5366)
                     /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     complement(4724..4743)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5537..6125
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6026..6045
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
       61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      181 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
      241 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      301 tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga ttacgaatta
      361 attcgagctc gcccgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
      421 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      481 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      541 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      601 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      661 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      721 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      781 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      841 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      901 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
      961 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag
     1021 acaccgggac cgatccagcc tccgcggccg ggaacggtgc attggaacgc ggattccccg
     1081 tgccaagagt gacgtaagta ccgcctatag agtctatagg cccaccccct tggcttcgtt
     1141 agaacgcggc tacaattaat acataacctt atgtatcata cacatacgat ttaggtgaca
     1201 ctatagaata acatccactt tgcctttctc tccacaggtg tccactccca ggtccaactg
     1261 cacctcggtt ctatcgattg aattcatgga ggcagcatcg gctgctttgc gtcgctgcct
     1321 gcttctcatc gtgttggtgg cggcggcgac gctgctcccg ggggcgaagg cattacagtg
     1381 tttctgccac ctctgtacaa aggacaattt tacttgtgag acagatggtc tctgctttgt
     1441 ctcagtcacc gagaccacag acaaagttat acacaatagc atgtgtatag ctgaaatcga
     1501 cctaattccc cgagacaggc catttgtttg tgcaccatct tcaaaaacag gggcagttac
     1561 gtattgctgc aatcaggatc actgcaataa aatagaactc ccaactacag gacctttttc
     1621 agaaaagcag tcagctggcc tcggtcctgt ggagctggca gctgtcattg ctggtccagt
     1681 ctgcttcgtc tgcattgcac ttatgctgat ggtctatatc tgccacaacg gatctatgcg
     1741 cactgtcatt caccaccgcg tgccaaatga agaggatccc tcactagatc gccctttcat
     1801 ttcagagggc accaccttaa aagatttaat ttatgatatg acaacatcag ggtctggatc
     1861 aggtttacca ctgcttgttc aaagaacaat tgcaagggac attgtgctac aagaaagcat
     1921 cggcaaaggt cggtttggag aagtttggcg aggcaaatgg cggggagaag aagttgccgt
     1981 gaagatattc tcttctagag aagaacgttc atggttccga gaggcagaga tttatcagac
     2041 tgtaatgtta cgccatgaaa atatcctggg gtttatagca gcagacaaca aagacaatgg
     2101 tacatggact cagctgtggt tggtgtcgga ttatcatgag catggatccc ttttcgatta
     2161 cttgaataga tacactgtta ctgtggaagg aatgatcaaa ctcgctctgt ccacggcaag
     2221 tggtcttgcc catcttcaca tggagattgt tggtacccaa ggaaaaccag ctattgccca
     2281 tagagatttg aaatcaaaga atatcttggt gaagaaaaat ggaacctgtt gtattgcaga
     2341 tttgggactt gctgtgagac atgattctgc cacagataca attgatattg ctccaaacca
     2401 cagagtaggc actaaaaggt atatggcccc tgaagttcta gatgattcca taaatatgaa
     2461 acattttgaa tccttcaaac gtgctgacat ctatgcaatg ggcttagtat tctgggaaat
     2521 cgctcgacgc tgttccattg gcggaatcca cgaagactac cagttgcctt actatgatct
     2581 tgtaccttct gatccatccg ttgaagaaat gagaaaagta gtttgtgaac agaagttaag
     2641 gccaaatatt cccaacagat ggcagagctg tgaggccttg agagtgatgg ccaaaattat
     2701 gagagaatgt tggtatgcca atggagcagc taggctgaca gctttgcgaa ttaaaaaaac
     2761 attgtcacag ctcagccaac aggaaggcat caaaatggtc gacatggact acaaggacga
     2821 cgacgacaag tgataagctt ggccgccatg gcccaacttg tttattgcag cttataatgg
     2881 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2941 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcg ggaattaatt
     3001 cggcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg
     3061 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3121 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3181 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3241 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3301 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3361 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3421 gttaacagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     3481 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     3541 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
     3601 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     3661 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3721 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3781 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3841 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     3901 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3961 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt
     4021 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4081 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     4141 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     4201 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4261 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     4321 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     4381 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4441 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4501 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4561 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4621 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4681 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4741 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4801 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4861 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4921 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4981 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5041 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5101 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5161 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5221 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5281 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5341 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5401 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5461 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5521 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5581 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5641 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5701 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5761 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5821 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5881 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5941 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6001 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6061 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6121 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6181 a
//
