LOCUS Exported 3961 bp ds-DNA circular SYN 18-SEP-2019 DEFINITION . ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3961) AUTHORS Clontech TITLE Direct Submission JOURNAL Exported Wednesday, Sep 18, 2019 from SnapGene 4.3.11 https://www.snapgene.com COMMENT Sequencing of the insert can be found at Seq004 FEATURES Location/Qualifiers source 1..3961 /organism="synthetic DNA construct" /lab_host="Escherichia coli" /mol_type="other DNA" promoter 1..451 /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 81..99 /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 117..135 /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 153..171 /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 189..207 /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 225..243 /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 261..279 /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 297..315 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" /note="color: #31849b" CDS 474..899 /codon_start=1 /product="First part of ProRIP till the internal inactivation region." /label=alpha-Chain_gePSI /note="This feature has 2 segments: 1: 474 .. 896 / #3366ff 2: 897 .. 899 / #3366ff" /translation="MKFTEIFPVEDANYPYSAFIASVRKDVIKHCTDHKGIFQPVLPPE KKVPELWLYTELKTRTSSITLAIRMDNLYLVGFRTPGGVWWEFGKDGDTHLLGDNPRWL GFGGRYQDLIGNKGLETVTMGRAEMTRAVNDLAKKKK" polyA_signal 1031..1112 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" /note="color: #a6acb3" rep_origin complement(1294..1882) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" CDS complement(2053..2913) /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: 2053 .. 2844 / #ccffcc 2: 2845 .. 2913 / #ccffcc / signal sequence Cleavage site after base 2844" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(2914..3018) /gene="bla" /label=AmpR promoter /note="color: #ffffff; direction: LEFT" polyA_signal 3051..3132 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" /note="color: #a6acb3" CDS complement(3252..3947) /codon_start=1 /product="Zoanthus green fluorescent protein" /label=ZsGreen1 /note="mammalian codon-optimized" /note="color: #05fd14" /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH AIASGSALP" ORIGIN 1 gcgatctgac ggttcactaa acgagctctg cttatatagg cctcccaccg tacacgccac 61 ctcgacatac tcgagtttac tccctatcag tgatagagaa cgtatgaaga gtttactccc 121 tatcagtgat agagaacgta tgcagacttt actccctatc agtgatagag aacgtataag 181 gagtttactc cctatcagtg atagagaacg tatgaccagt ttactcccta tcagtgatag 241 agaacgtatc tacagtttac tccctatcag tgatagagaa cgtatatcca gtttactccc 301 tatcagtgat agagaacgta taagctttag gcgtgtacgg tgggcgccta taaaagcaga 361 gctcgtttag tgaaccgtca gatcgcctgg agcaattcca caacactttt gtcttatacc 421 aactttccgt accacttcct accctcgtaa agtcgacacc ggggcccaga tctatgaagt 481 tcactgaaat cttccccgtg gaggacgcga actaccctta cagcgccttc atcgcgtcgg 541 tccggaaaga cgtgatcaaa cactgcaccg accataaagg gatcttccag cccgtgctgc 601 caccggagaa gaaggtcccg gagctatggt tgtacacaga gctcaaaact aggaccagct 661 ccatcacgct cgccatacgc atggacaacc tgtacctcgt gggcttcagg accccgggcg 721 gggtgtggtg ggagttcggc aaggacggcg acacccacct cctcggcgac aaccccaggt 781 ggctcggctt cggcggcagg taccaggacc tcatcggcaa caagggtctg gagaccgtca 841 ccatgggccg cgccgaaatg accagggccg tcaacgacct ggcgaagaag aagaagtgag 901 cggccgccgg cgatatctcc agaggatcat aatcagccat accacatttg tagaggtttt 961 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat 1021 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 1081 aaatttcaca aataaagcat ttttttcact gccccgagct tcctcgctca ctgactcgct 1141 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 1201 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 1261 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 1321 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 1381 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 1441 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg 1501 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 1561 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 1621 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 1681 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt 1741 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 1801 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 1861 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 1921 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 1981 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 2041 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 2101 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 2161 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 2221 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 2281 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 2341 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 2401 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 2461 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 2521 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 2581 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 2641 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 2701 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 2761 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 2821 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 2881 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 2941 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 3001 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg gcagtgaaaa 3061 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc 3121 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg 3181 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcct 3241 ctagactgca gtcagggcaa ggcggagccg gaggcgatgg cgtgctcggt caggtgccac 3301 ttctggttct tggcgtcgct gcggtcctcg cgggtcagct tgtgctggat gaagtgccag 3361 tcgggcatct tgcggggcac ggacttggcc ttgtacacgg tgtcgaactg gcagcgcaag 3421 cggccaccgt ccttcagcag caggtacatg ctcacgtcgc ccttcaagat gccctgcttg 3481 ggcacgggga tgatcttctc gcaggagggc tcccagttgt cggtcatctt cttcatcacg 3541 gggccgtcgg cggggaagtt cacgccgtag aacttggact cgtggtacat gcagttctcc 3601 tccacgctca cggtgatgtc ggcgttgcag atgcacacgg cgccgtcctc gaacaggaag 3661 gagcggtccc aggtgtagcc ggcggggcag gagttcttga agtagtcgac gatgtcctgg 3721 gggtactcgg tgaacacgcg gttgccgtac atgaaggcgg cggacaagat gtcctcggcg 3781 aagggcaagg ggccgccctc caccacgcac aggttgatgg cctgcttgcc cttgaagggg 3841 tagccgatgc cctcgccggt gatcacgaac ttgtggccgt ccacgcagcc ctccatgcgg 3901 tacttcatgg tcatctcctt ggtcaggccg tgcttggact gggccatggt gaattctcca 3961 g //