LOCUS Exported 6256 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCMV5B-TGFbeta receptor I K232R 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6256 /organism="synthetic DNA construct" /mol_type="other DNA" 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(941..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 5878..5897 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" 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(6084..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 //