LOCUS pET_blank_Kan 4980 bp DNA circular SYN 06-DEC-2024 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4980) AUTHORS siwong TITLE Direct Submission JOURNAL Exported Dec 6, 2024 from SnapGene 8.0.0 https://www.snapgene.com FEATURES Location/Qualifiers source 1..4980 /mol_type="other DNA" /organism="synthetic DNA construct" misc_feature 1..88 /label=Custom promoter-RBS promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 20..44 /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)." RBS 59..81 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" misc_feature 75..88 /label=T7-lacO (no RBS) misc_feature 88^89 /label=T7-lacO-RBS terminator 158..205 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 242..697 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" CDS complement(790..1605) /codon_start=1 /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" rep_origin 1727..2315 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature 2501..2643 /label=bom /note="basis of mobility region from pBR322" CDS complement(2745..2936) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(3508..4590) /codon_start=1 /gene="lacI" /product="lac repressor" /label=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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" promoter complement(4591..4668) /gene="lacI" /label=lacI promoter ORIGIN 1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt 61 tgtttaactt taagaaggag atatacattt aatacctagg ctgctaaaca aagcccgaaa 121 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc 181 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg 241 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 301 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 361 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt 421 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg 481 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt 541 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta 601 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt 661 aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt tcggggaaat 721 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg 781 aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg 841 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag 901 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc 961 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg 1021 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc 1081 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat 1141 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac 1201 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga 1261 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa 1321 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt 1381 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg 1441 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga 1501 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt 1561 taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt 1621 actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa cgtgagtttt 1681 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt 1741 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt 1801 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga 1861 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag 1921 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata 1981 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg 2041 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga 2101 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca 2161 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 2221 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 2281 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac 2341 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt 2401 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga 2461 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc 2521 ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta caatctgctc 2581 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct 2641 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca 2701 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg 2761 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat 2821 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat 2881 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat 2941 gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg 3001 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt 3061 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag 3121 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag 3181 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg 3241 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt 3301 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct 3361 agccgggtcc tcaacgacag gagcacgatc atgctagtca tgccccgcgc ccaccggaag 3421 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg 3481 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 3541 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 3601 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg 3661 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg 3721 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac 3781 taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag 3841 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg 3901 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg 3961 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga 4021 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac 4081 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga 4141 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg 4201 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac 4261 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc 4321 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag 4381 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg 4441 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga 4501 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc 4561 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg 4621 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac 4681 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga 4741 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca 4801 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc 4861 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg 4921 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat cgatctcgat cccgcgaaat //