LOCUS Exported 6387 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCMV5B-TFGbeta receptor II K277R 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6387 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 148..283 /label=SV40 ori /note="SV40 origin of replication" primer_bind 210..229 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" 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(416..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(1529..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 6355..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 //