LOCUS pTE5422_[pSB0101_1_PT7-4bp-tetO_B0035-sfGFP_1xpepper_T7term] 2797 bp DNA circular UNA 26-OCT-2024 DEFINITION Lvl1 MoClo vector, PT7-tetO driving expression of sfGFP and Pepper in one transcript, RBS: B0035. ACCESSION urn.local.BarthelS.46m-idhjfyu VERSION urn.local.BarthelS.46m-idhjfyu KEYWORDS . SOURCE ORGANISM . FEATURES Location/Qualifiers CDS 2..718 /ApEinfo_fwdcolor="#34FF03" /ApEinfo_revcolor="#34FF03" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}" /modified_by="BarthelS" /label="sfGFP" misc_RNA 723..773 /modified_by="BarthelS" /label="Pepper" terminator 780..826 /note="transcription terminator for bacteriophage T7 RNA polymerase" /label="T7 terminator" terminator complement(854..950) /label="AmpR Terminator" CDS complement(951..1808) /label="AmpR" primer_bind complement(956..978) /Tm=57 /Sequence="GATAGGTGCCTCACTGATTAAGC" /Description="" /label="oSB0022_lvl1_seq_rev" promoter complement(1809..1939) /label="AmpR Promoter" rep_origin complement(1940..2703) /snapgene_direction="LEFT" /label="ColE1" primer_bind 2223..2246 /created_by="Hoffmann-Becking" /Sequence="CCTCGCTCTGCTAATCCTGTTACC" /Tm="62.8" /%GC="54.2" /Hairpin_Tm="None" /Self_Dimer_Tm="None" /modified_by="BarthelS" /note="Derived using Geneious Prime 2024.0.7 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pTE5415 [pSB0112_0_04_phi29 DNAP wt]" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; oMHB011_qPCR_rev (pTE5415 [pSB0112_0_04_phi29 DNAP wt])" /label="oMHB011_qPCR_rev" primer_bind complement(2362..2382) /created_by="Hoffmann-Becking" /Sequence="GGTGTAGGTCGTTCGCTCCAA" /Tm="62.4" /%GC="57.1" /Hairpin_Tm="None" /Self_Dimer_Tm="8.3" /modified_by="BarthelS" /note="Derived using Geneious Prime 2024.0.7 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pTE5415 [pSB0112_0_04_phi29 DNAP wt]" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; oMHB010_qPCR_fw (pTE5415 [pSB0112_0_04_phi29 DNAP wt])" /label="oMHB010_qPCR_fw" primer_bind 2629..2646 /Tm=58 /Sequence="CGGTTCCTGGCCTTTTGC" /Description="" /label="oSB0021_lvl1_seq_fwd" promoter 2731..2747 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" misc_feature 2748..2751 /label="4bp spacer" Operator 2752..2770 /bound_moiety="tetracycline repressor TetR" /gene="tetO" /note="bacterial operator O₂ for the tetR and tetA genes" /modified_by="BarthelS" /label="tet operator" RBS 2779..2792 /created_by="BarthelS" /label="B0035" ORIGIN 1 aatgcgtaaa ggcgaagagc tgttcactgg tgtcgtccct attctggtgg aactggatgg 61 tgatgtcaac ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg caactaatgg 121 taaactgacg ctgaagttca tctgtactac tggtaaactg ccggttcctt ggccgactct 181 ggtaacgacg ctgacttatg gtgttcagtg ctttgctcgt tatccggacc atatgaagca 241 gcatgacttc ttcaagtccg ccatgccgga aggctatgtg caggaacgca cgatttcctt 301 taaggatgac ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg ataccctggt 361 aaaccgcatt gagctgaaag gcattgactt taaagaggac ggcaatatcc tgggccataa 421 gctggaatac aattttaaca gccacaatgt ttacatcacc gccgataaac aaaaaaatgg 481 cattaaagcg aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc agctggctga 541 tcactaccag caaaacactc caatcggtga tggtcctgtt ctgctgccag acaatcacta 601 tctgagcacg caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc atatggttct 661 gctggagttc gtaaccgcag cgggcatcac gcatggtatg gatgaactgt acaaataagc 721 ttggccccaa tcgtggcgtg tcggcctgct tcggcaggca ctggcgccgg gccgcttaat 781 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgcgct tgttggagac 841 gagctgcggc cgcgattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 901 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 961 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 1021 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 1081 ataccgcggg acccacgctc accggctcca gatttatcag caataaacca gccagccgga 1141 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 1201 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 1261 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc 1321 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc 1381 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca 1441 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag 1501 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg 1561 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa 1621 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 1681 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga 1741 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga 1801 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 1861 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 1921 ccccgaaaag tgccacctgt catgaccaaa atcccttaac gtgagttttc gttccactga 1981 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta 2041 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa 2101 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact 2161 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca 2221 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt 2281 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg 2341 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag 2401 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta 2461 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat 2521 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg 2581 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc 2641 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac 2701 cgtgcggccg caacacgtct cgagtaggag taatacgact cactataggg atccctatca 2761 gtgatagaga tactagagat taaagaggag aataatc //