LOCUS Exported 6619 bp ds-DNA circular SYN 23-JUL-2019 DEFINITION . ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6619) AUTHORS . TITLE Direct Submission JOURNAL Exported Tuesday, Jul 23, 2019 from SnapGene 4.3.10 https://www.snapgene.com FEATURES Location/Qualifiers source 1..6619 /organism="synthetic DNA construct" /mol_type="other DNA" /note="color: #ffffff" repeat_region 1..141 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" /note="color: #ffe4c4" polyA_signal 278..359 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" /note="color: #a6acb3" CDS complement(485..595) /codon_start=1 /product="Amino acid sequence taken from the book ""Synthetic Protein Switches Methods and Protocols""-Chapter 15-Fig 1a. Book: SSN 1064-3745 ISSN 1940-6029 (electronic)Methods in Molecular Biology ISBN 978-1-4939-6938-8 ISBN 978-1-4939-6940-1 (eBook)DOI 10.1007/978-1-4939-6940-1 Amino acid sequence was codon optimized for Mus musculus." /label=ODC36 /note="color: #ff6600" /translation="GITHGMDELYKLPMSCAQESITSLYKKAGSENLYFQ" CDS complement(596..904) /codon_start=1 /label=beta-Chain_gePSI /note="color: #3366ff" /translation="MAADPQADTKSKLVKLVVMVCEGLRFNTVSRTVDAGFNSQHGVTL TVTQGKQVQKWDRISKAAFEWADHPTAVIPDMQKLGIKDKNEAARIVALVKNQTTAAA" promoter 919..1369 /label=TRE3G BI promoter /note="3rd-generation Tet-responsive bidirectional promoter that can be activated by binding of Tet-On 3G" /note="color: #ffffff; direction: BOTH" protein_bind 999..1017 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" /note="color: #31849b" protein_bind 1035..1053 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" /note="color: #31849b" protein_bind 1071..1089 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" /note="color: #31849b" protein_bind 1107..1125 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" /note="color: #31849b" protein_bind 1143..1161 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" /note="color: #31849b" protein_bind 1179..1197 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" /note="color: #31849b" protein_bind 1215..1233 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" /note="color: #31849b" CDS 1392..1817 /codon_start=1 /product="First part of ProRIP till the internal inactivation region." /label=alpha-Chain_gePSI /note="This feature has 2 segments: 1: 1392 .. 1814 / #3366ff 2: 1815 .. 1817 / #3366ff" /translation="MKFTEIFPVEDANYPYSAFIASVRKDVIKHCTDHKGIFQPVLPPE KKVPELWLYTELKTRTSSITLAIRMDNLYLVGFRTPGGVWWEFGKDGDTHLLGDNPRWL GFGGRYQDLIGNKGLETVTMGRAEMTRAVNDLAKKKK" polyA_signal 1949..2030 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" /note="color: #a6acb3" CDS 2115..2769 /label=CMV /note="color: #993366; direction: RIGHT" enhancer 2141..2520 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" /note="color: #ffffff" promoter 2521..2724 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" /note="color: #ffffff; direction: RIGHT" promoter 2769..2787 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" /note="color: #ffffff; direction: RIGHT" CDS 2833..3552 /codon_start=1 /product="Aequorea coerulescens GFP" /label=AcGFP1 /note="mammalian codon-optimized" /note="color: #05fd14" /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF GFVTAAAITHGMDELYK" misc_feature 3570..3572 /label=Belongs to bGH /note="color: #a6acb3" polyA_signal 3573..3797 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" /note="color: #a6acb3" repeat_region 3882..4022 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" /note="color: #ffe4c4" rep_origin 4097..4552 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /note="color: #ffff00" promoter 4834..4938 /gene="bla" /label=AmpR promoter /note="color: #ffffff; direction: RIGHT" CDS 4939..5799 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /note="This feature has 2 segments: 1: 4939 .. 5007 / #ccffcc / signal sequence 2: 5008 .. 5799 / #ccffcc Cleavage site after base 5007" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 5947..6614 /direction=RIGHT /label=pUCori /note="color: #ffff00" rep_origin 5970..6558 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 121 actccatcac taggggttcc tgcggccaat tcagtcgata actataacgg tcctaaggta 181 gcgatttaaa tacgcgctct cttaaggtag ccccgggacg cgtcaattgc atgaagaatc 241 tgcttagggt taggcgtttt gctgccacct gacgtcggca gtgaaaaaaa tgctttattt 301 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta 361 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt 421 aaagcaagta aaacctctac aaatgtggta tggctgatta tgatcctcta gactgcagac 481 tagttcattg aaagtacaag ttctctgaac cagctttttt gtagagagat gttatgcttt 541 cctgggcaca agacattggc agcttgtaaa gttcgtccat tccgtgtgtt atcccagccg 601 ccgcagtagt ttgattctta acgagcgcaa cgatcctcgc tgcttcgttc ttatccttga 661 tgccaagctt ctgcatgtcg gggatcacag cggtggggtg gtcagcccac tcgaaggccg 721 ccttggagat cctgtcccac ttctgcacct gcttcccctg cgtcacggtc aaggtcaccc 781 cgtgctggct gttgaacccc gcgtccaccg tgcgggacac ggtgttgaac cgcagcccct 841 cgcacaccat gaccaccagc ttcaccagct tgctcttcgt gtcggcctgt gggtcagccg 901 ccatggtgaa ttctccaggc gatctgacgg ttcactaaac gagctctgct tatataggcc 961 tcccaccgta cacgccacct cgacatactc gagtttactc cctatcagtg atagagaacg 1021 tatgaagagt ttactcccta tcagtgatag agaacgtatg cagactttac tccctatcag 1081 tgatagagaa cgtataagga gtttactccc tatcagtgat agagaacgta tgaccagttt 1141 actccctatc agtgatagag aacgtatcta cagtttactc cctatcagtg atagagaacg 1201 tatatccagt ttactcccta tcagtgatag agaacgtata agctttaggc gtgtacggtg 1261 ggcgcctata aaagcagagc tcgtttagtg aaccgtcaga tcgcctggag caattccaca 1321 acacttttgt cttataccaa ctttccgtac cacttcctac cctcgtaaag tcgacaccgg 1381 ggcccagatc tatgaagttc actgaaatct tccccgtgga ggacgcgaac tacccttaca 1441 gcgccttcat cgcgtcggtc cggaaagacg tgatcaaaca ctgcaccgac cataaaggga 1501 tcttccagcc cgtgctgcca ccggagaaga aggtcccgga gctatggttg tacacagagc 1561 tcaaaactag gaccagctcc atcacgctcg ccatacgcat ggacaacctg tacctcgtgg 1621 gcttcaggac cccgggcggg gtgtggtggg agttcggcaa ggacggcgac acccacctcc 1681 tcggcgacaa ccccaggtgg ctcggcttcg gcggcaggta ccaggacctc atcggcaaca 1741 agggtctgga gaccgtcacc atgggccgcg ccgaaatgac cagggccgtc aacgacctgg 1801 cgaagaagaa gaagtgagcg gccgccggcg atatctccag aggatcataa tcagccatac 1861 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa 1921 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa 1981 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc cccgagcttc 2041 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 2101 aaagggctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt 2161 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt 2221 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg 2281 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg 2341 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt 2401 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg 2461 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg 2521 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt 2581 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac 2641 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg 2701 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt 2761 atcgaaatta atacgactca ctatagggag acccaagctg gctagcgttt aaacttaagc 2821 ttggtaccga gcatggtgag caagggcgcc gagctgttca ccggcatcgt gcccatcctg 2881 atcgagctga atggcgatgt gaatggccac aagttcagcg tgagcggcga gggcgagggc 2941 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcctgtg 3001 ccctggccca ccctggtgac caccctgagc tacggcgtgc agtgcttctc acgctacccc 3061 gatcacatga agcagcacga cttcttcaag agcgccatgc ctgagggcta catccaggag 3121 cgcaccatct tcttcgagga tgacggcaac tacaagtcgc gcgccgaggt gaagttcgag 3181 ggcgataccc tggtgaatcg catcgagctg accggcaccg atttcaagga ggatggcaac 3241 atcctgggca ataagatgga gtacaactac aacgcccaca atgtgtacat catgaccgac 3301 aaggccaaga atggcatcaa ggtgaacttc aagatccgcc acaacatcga ggatggcagc 3361 gtgcagctgg ccgaccacta ccagcagaat acccccatcg gcgatggccc tgtgctgctg 3421 cccgataacc actacctgtc cacccagagc gccctgtcca aggaccccaa cgagaagcgc 3481 gatcacatga tctacttcgg cttcgtgacc gccgccgcca tcacccacgg catggatgag 3541 ctgtacaagt gaaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt 3601 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc 3661 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg 3721 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga 3781 tgcggtgggc tctatggctt ctgaggcgga aagaaccaga tcctctctta aggtagcatc 3841 gagatttaaa ttagggataa cagggtaatg gcgcgggccg caggaacccc tagtgatgga 3901 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc 3961 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca 4021 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg 4081 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 4141 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 4201 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct 4261 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat 4321 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 4381 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc 4441 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg 4501 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc 4561 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca 4621 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg 4681 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga 4741 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct 4801 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc 4861 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa 4921 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt 4981 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct 5041 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc 5101 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta 5161 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac 5221 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc 5281 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac 5341 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg 5401 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac 5461 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc 5521 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt 5581 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga 5641 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc 5701 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag 5761 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca 5821 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc 5881 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca 5941 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc 6001 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta 6061 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt 6121 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc 6181 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg 6241 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg 6301 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag 6361 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc 6421 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat 6481 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg 6541 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc 6601 tggccttttg ctcacatgt //