LOCUS Exported 5108 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION The tet-responsive reporter pBI-MCS-EGFP (Addgene plasmid 16542) was modified to express YFP instead of GFP. ACCESSION . VERSION . KEYWORDS pBI-MCS-EYFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5108) AUTHORS Daringer NM, Dudek RM, Schwarz KA, Leonard JN TITLE Modular Extracellular Sensor Architecture for Engineering Mammalian Cell-based Devices. JOURNAL ACS Synth Biol. 2014 Mar 11. PUBMED 24611683 REFERENCE 2 (bases 1 to 5108) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5108 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 135..405 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" promoter 438..475 /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" intron 657..1229 /label=-beta-globin intron /note="intron from rabbit beta-globin gene" primer_bind complement(1361..1380) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 1426..1481 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(1480..1499) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(1872..1889) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2043..2631) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2123..2142) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2802..3662) /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 3425..3444 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3663..3767) /gene="bla" /label=AmpR promoter primer_bind 3835..3853 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" polyA_signal 4198..4332 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4235..4254) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4289..4308 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" CDS complement(4369..5088) /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4414..4435) /label=EGFP-C /note="EGFP, forward primer" primer_bind 4763..4782 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5022..5043 /label=EGFP-N /note="EGFP, reverse primer" ORIGIN 1 tgcaggaatt ccgggccgcg gaggctggat cggtcccggt gtcttctatg gaggtcaaaa 61 cagcgtggat ggcgtctcca ggcgatctga cggttcacta aacgagctct gcttatatag 121 gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc 181 tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa 241 gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc 301 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag 361 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag 421 ctcggtaccc gggtcgagta ggcgtgtacg gtgggaggcc tatataagca gagctcgttt 481 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca 541 ccgggaccga tccagcctcc gcggccccga attcgagctc ggtacccggg gatcctctag 601 tcagctgacg cgtgctagcg cggcctcgac gatatctcta gactgagaac ttcagggtga 661 gtttggggac ccttgattgt tctttctttt tcgctattga aaaattcatg ttatatggag 721 ggggcaaagt tttcagggtg ttgtttagaa tgggaagatg tcccttgtat caccatggac 781 cctcatgata attttgtttc tttcactttc tactctgttg acaaccattg tctcctctta 841 ttttcttttc attttctgta acttttttcg ttaaacttta gcttgcattt gtaacgaatt 901 tttaaattca ctttcgttta tttgtcagat tgtaagtact ttctctaatc actttttttt 961 caaggcaatc agggtaatta tattgtactt cagcacagtt ttagagaaca attgttataa 1021 ttaaatgata aggtagaata tttctgcata taaattctgg ctggcgtgga aatattctta 1081 ttggtagaaa caactacatc ctggtaatca tcctgccttt ctctttatgg ttacaatgat 1141 atacactgtt tgagatgagg ataaaatact ctgagtccaa accgggcccc tctgctaacc 1201 atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt gttgtgctgt 1261 ctcatcattt tggcaaagaa ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg 1321 ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa 1381 attatgggga catcatgaag ccccttgagc atctgacttc tgggtaataa aggaaattta 1441 ttttcattgc aatagtgtgt gggaattttt tgtgtctctc actcggaagg acatatggga 1501 gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca acatatgcca 1561 tatgctggct gccatgaaca aaggtggcta taaagaggtc atcagtatat gaaacagccc 1621 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata 1681 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta 1741 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatga 1801 actcgactgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 1861 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 1921 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 1981 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 2041 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 2101 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 2161 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 2221 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 2281 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 2341 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 2401 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 2461 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt 2521 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 2581 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 2641 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 2701 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 2761 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 2821 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 2881 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 2941 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 3001 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 3061 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 3121 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 3181 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 3241 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 3301 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 3361 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 3421 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 3481 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 3541 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 3601 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 3661 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 3721 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 3781 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg 3841 cgtatcacga ggccctttcg tcttcactcg acgctggtcg agctgatact tcccgtccgc 3901 caggggacat gccggcgatg ctgaaggtcg cgcgcattcc cgatgaagag gccggttacc 3961 gcctgttgac ctggtgggac gggcagggcg ccgcccgagt cttgcctcgg cggcgggcgc 4021 tctgctcatg gagcgcgcgt ccggggacct tgcacagata gcgtggtccg gccagacgac 4081 gaggcttgca ggatcataat cagccatacc acatttgtag aggttttact tgctttaaaa 4141 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac 4201 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 4261 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 4321 catgtctgga tcctctagaa ctaggtcgac aagcttgata tcgaattctt acttgtacag 4381 ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc 4441 gcgcttctcg ttggggtctt tgctcagggc ggactggtag ctcaggtagt ggttgtcggg 4501 cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac 4561 gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt tcttctgctt 4621 gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat 4681 gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc 4741 ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg 4801 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc 4861 ggggtagcgg gcgaagcact gcaggccgta gccgaaggtg gtcacgaggg tgggccaggg 4921 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc 4981 gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac 5041 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatac tagttctaga 5101 gcggccgc //