LOCUS Exported 8880 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Amber suppression for incorporating 3-aminotyrosine to proteins in mammalian cells. ACCESSION . VERSION . KEYWORDS pMAH-EcaYRS SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8880) AUTHORS Zhang S, Ai HW TITLE A general strategy to red-shift green fluorescent protein-based biosensors. JOURNAL Nat Chem Biol. 2020 Sep 14. pii: 10.1038/s41589-020-0641-7. doi: 10.1038/s41589-020-0641-7. PUBMED 32929278 REFERENCE 2 (bases 1 to 8880) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8880 /organism="synthetic DNA construct" /mol_type="other DNA" promoter complement(111..201) /label=H1 promoter /note="human H1 RNA promoter" promoter complement(321..561) /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" promoter complement(850..940) /label=H1 promoter /note="human H1 RNA promoter" promoter complement(1060..1300) /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" enhancer 1559..1862 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1863..2066 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2017..2037 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2063..2087 /label=LNCX /note="Human CMV promoter, forward primer" promoter 2297..2315 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 2297..2314 /label=SP6 /note="SP6 promoter, forward primer" primer_bind 2328..2347 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 2408..2417 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(4320..4337) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4326..4550 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4596..5024 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4683..4702) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4893..4914 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5033..5053) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5038..5367 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5218..5353 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5280..5299 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5416..6441 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" polyA_signal 6571..6692 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6608..6627) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6662..6681 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6741..6757) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6741..6757) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6754..6776) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6765..6781 /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(6789..6819) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6834..6855 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6972..6989) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7143..7731) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7223..7242) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7902..8762) /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 8525..8544 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8763..8867) /gene="bla" /label=AmpR promoter ORIGIN 1 ctcgactcgc tagtgttcca aaaaatggtg ggggacggat tcgaaccgcc gaacccaaag 61 ggagcggatt tagagtccgc ggcgtttggc cacttcgcta ccccaccaga gtggtctcat 121 acagaactta taagattccc aaatccaaag acatttcacg tttatggtga tttcccagaa 181 cacatagcga catgcaaata tgaattgatg actgcctgat ggagaaaaaa tggtggggga 241 cggattcgaa ccgccgaacc caaagggagc ggatttagag tccgcggcgt ttggccactt 301 cgctacccca ccggtgtttc gtcctttcca caagatatat aaagccaaga aatcgaaata 361 ctttcaagtt acggtaagca tatgatagtc cattttaaaa cataatttta aaactgcaaa 421 ctacccaaga aattattact ttctacgtca cgtattttgt actaatatct ttgtgtttac 481 agtcaaatta attctaatta tctctctaac agccttgtat cgtatatgca aatatgaagg 541 aatcatggga aataggccct cttcctgccc gaccttcgcg atgtacgggc cagatatacg 601 ccaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta 661 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc 721 aatatgaccg ccatgttggc tcgactcgct agtgttccaa aaaatggtgg gggacggatt 781 cgaaccgccg aacccaaagg gagcggattt agagtccgcg gcgtttggcc acttcgctac 841 cccaccagag tggtctcata cagaacttat aagattccca aatccaaaga catttcacgt 901 ttatggtgat ttcccagaac acatagcgac atgcaaatat gaattgatga ctgcctgatg 961 gagaaaaaat ggtgggggac ggattcgaac cgccgaaccc aaagggagcg gatttagagt 1021 ccgcggcgtt tggccacttc gctaccccac cggtgtttcg tcctttccac aagatatata 1081 aagccaagaa atcgaaatac tttcaagtta cggtaagcat atgatagtcc attttaaaac 1141 ataattttaa aactgcaaac tacccaagaa attattactt tctacgtcac gtattttgta 1201 ctaatatctt tgtgtttaca gtcaaattaa ttctaattat ctctctaaca gccttgtatc 1261 gtatatgcaa atatgaagga atcatgggaa ataggccctc ttcctgcccg accttcgcga 1321 tgtacgggcc agatatacgc caatattggc cattagccat attattcatt ggttatatag 1381 cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca 1441 tttatattgg ctcatgtcca atatgaccgc catgttggct cgagcattga ttattgacta 1501 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 1561 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 1621 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 1681 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 1741 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 1801 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 1861 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat 1921 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 1981 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac 2041 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc 2101 atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc cgcggccggg 2161 aacggtgcat tggaacgcgg attccccgtg ccaagagtga cgtaagtacc gcctatagag 2221 tctataggcc cacccccttg gcttcgttag aacgcggcta caattaatac ataaccttat 2281 gtatcataca catacgattt aggtgacact atagaataac atccactttg cctttctctc 2341 cacaggtgtc cactcccagg tccaactgca cggaagcttg cccccaggtc caactgcacg 2401 gaagcttgcc accatggcaa gcagtaactt gattaaacaa ttgcaagagc gggggctggt 2461 agcccaggtg acggacgagg aagcgttagc agagcgactg gcgcaaggcc cgatcgcgct 2521 cctttgcggc ttcgatccta ccgctgacag cttgcatttg gggcatcttg ttccattgtt 2581 atgcctgaaa cgcttccagc aggcgggcca caagccggtt gcgctggtag gcggcgcgac 2641 gggtctgatt ggcgacccga gcttcaaagc tgccgagcgt aagctgaaca ccgaagaaac 2701 tgttcaggag tgggtggaca aaatccgtaa gcaggttgcc ccgttcctcg atttcgactg 2761 tggagaaaac tctgctatcg cggcgaacaa ctatgactgg ttcggcaata tgaatgtgct 2821 gaccttcctg cgcgatattg gcaaacactt ctccgttaac cagatgatca acaaagaagc 2881 ggttaagcag cgtctcaacc gtgaagatca ggaggttaaa aagagtaacg taacggttta 2941 cgttgatgag aaactgaaag atataaggcc ttatggagtt tacgcaatag ttgaaggttt 3001 aaggctcgac gaagattctt taagtcaaat gattcagcta caagaaaaga tagcccttac 3061 atttggaaga agaaggagag aagtggccat aggaatcttc gattttgata agattaagcc 3121 acctatttac tataaagccg ccgaaaaaac tgaaaagttt gcccccctgg gctataaaga 3181 ggaaatgact ctagaggaga tccttgaaaa gcatgaaaag ggaagggagt atgggcacct 3241 tataaaggat aaacaatttt atccactact tattgacagc gaggggaatg tgctctccat 3301 gccgccaata atcaactccg agtttacggg aagagtaaca acggatacga aaaatgtctt 3361 catagatgtc acgggatgga aacttgagaa ggtaatgctt gcccttaatg tcatggtaac 3421 tgcattagca gagcgtggag gtaaaataag gagcgttagg gttgtctaca aggacttcga 3481 aattgaaacc ccaggctccg cctccggccc cgcctccgcc gggatttcgt tcactgagtt 3541 ttcctacaac ctgttgcagg gttatgactt cgcctgtctg aacaaacagt acggtgtggt 3601 gctgtctatt ggtggttctg accagtgggg taacatcact tctggtatcg acctgacccg 3661 tcgtctgcat cagaatcagg tgtttggcct gaccgttccg ctgatcacta aagcagatgg 3721 caccaaattt ggtaaaactg aaggcggcgc agtctggttg gatccgaaga aaaccagccc 3781 gtacaaattc taccagttct ggatcaacac tgcgcgggcc gacgtttacc gcttcctgaa 3841 gttcttcacc tttatgagca ttgaagagat caacgccctg gaagaagaag ataaaaacag 3901 cggtaaagca ccgcgcgccc agtatgtact ggcggagcag gtgactcgtc tggttcacgg 3961 tgaagaaggt ttacaggcgg caaaacgtat taccgaatgc ctgttcagcg gttctttgag 4021 tgcgctgagt gaagcggact tcgaacagct ggcgcaggac ggcgtaccga tggttgagat 4081 ggaaaagggc gcagacctga tgcaggcact ggtcgattct gaactgcaac cttcccgtgg 4141 tcaggcacgt aaaactatcg cctccaatgc catcaccatt aacggtgaaa aacagtccga 4201 tcctgaatac ttctttaaag aagaagatcg tctgtttggt cgttttacct tactgcgtcg 4261 cggtaaaaag aattactgtc tgatttgctg gaaagggccc gtttaaaccc gctgatcagc 4321 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 4381 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 4441 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 4501 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc 4561 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag 4621 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 4681 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 4741 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 4801 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 4861 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 4921 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta 4981 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg 5041 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 5101 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt 5161 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc 5221 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 5281 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc 5341 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga 5401 tctgatcagc acgtgatgaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg 5461 atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt 5521 gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat 5581 ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg 5641 gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca 5701 cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc 5761 gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca 5821 ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct 5881 gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg 5941 caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg 6001 cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt 6061 gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg 6121 aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag 6181 cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat 6241 cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca 6301 atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc 6361 gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact 6421 cgtccgaggg caaaggaata gcacgtgcta cgagatttcg attccaccgc cgccttctat 6481 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg 6541 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac 6601 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 6661 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc 6721 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 6781 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 6841 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 6901 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 6961 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 7021 tcagctcact caaagacggt aatacggtta tccacagaat caggggataa cgcaggaaag 7081 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 7141 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 7201 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 7261 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 7321 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 7381 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 7441 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 7501 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 7561 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt 7621 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 7681 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 7741 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 7801 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 7861 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 7921 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 7981 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 8041 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 8101 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 8161 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 8221 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 8281 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 8341 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 8401 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 8461 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 8521 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 8581 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 8641 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 8701 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 8761 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 8821 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga //