LOCUS Exported 6409 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pAL101 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6409) AUTHORS Levskaya A, Weiner OD, Lim WA, Voigt CA TITLE Spatiotemporal control of cell signalling using a light-switchable protein interaction. JOURNAL Nature. 2009 Sep 13. ():. PUBMED 19749742 REFERENCE 2 (bases 1 to 6409) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6409 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1207..1926 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1252..1273) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1513..1532) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1860..1881 /label=EGFP-C /note="EGFP, forward primer" promoter complement(1961..1979) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(1962..1979) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(1999..2016) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2005..2229 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2275..2703 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2362..2381) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2572..2593 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2712..2732) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2717..3047 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2898..3033 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2960..2979 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3114..3908 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3168..3187) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3778..3797 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4082..4203 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4119..4138) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4173..4192 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4252..4268) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4252..4268) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4265..4287) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4276..4292 /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(4300..4330) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4345..4366 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4483..4500) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4654..5242) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4734..4753) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5413..6273) /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 6036..6055 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6274..6378) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc 901 atgatgttct taccaaccga ttattgttgc aggttaagcg atcaagagta tatggagctt 961 gtgtttgaga atggccagat tcttgcaaag ggccaaagat ccaacgtttc tctgcataat 1021 caacgtacca aatcgatcat ggatttgtat gaggcagagt ataacgagga tttcatgaag 1081 agtatcatcc atggtggtgg tggtgccatc acaaatctcg gggacacgca ggttgttcca 1141 caaagtcatg ttgctgctgc ccatgaaaca aacatgttgg aaagcaataa acatgttgac 1201 ggatccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag 1261 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc 1321 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg 1381 cccaccctcg tgaccacctt cggctacggc ctgcagtgct tcgcccgcta ccccgaccac 1441 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc 1501 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac 1561 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg 1621 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag 1681 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag 1741 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac 1801 aaccactacc tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac 1861 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac 1921 aagtaagcgg ccgctcgagc atgcatctag agggccctat tctatagtgt cacctaaatg 1981 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc 2041 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 2101 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 2161 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg 2221 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc 2281 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 2341 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 2401 ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt 2461 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 2521 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 2581 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga 2641 ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 2701 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccaggcag 2761 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 2821 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 2881 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 2941 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat 3001 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag 3061 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg 3121 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 3181 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 3241 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg 3301 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 3361 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 3421 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 3481 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 3541 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 3601 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg 3661 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 3721 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 3781 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 3841 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 3901 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc 3961 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg 4021 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc 4081 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 4141 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 4201 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct 4261 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 4321 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 4381 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 4441 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 4501 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 4561 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 4621 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 4681 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 4741 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 4801 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg 4861 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 4921 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 4981 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 5041 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt 5101 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 5161 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 5221 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 5281 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 5341 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 5401 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 5461 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 5521 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 5581 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 5641 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 5701 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 5761 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 5821 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 5881 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 5941 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 6001 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 6061 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 6121 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 6181 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 6241 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 6301 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 6361 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc //