LOCUS Exported 5356 bp DNA circular SYN 10-SEP-2025 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5356) AUTHORS 11111111 TITLE Direct Submission JOURNAL Exported Sep 10, 2025 from SnapGene 6.0.2 https://www.snapgene.com FEATURES Location/Qualifiers source 1..5356 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 265..283 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 284..308 /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)." CDS 353..418 /codon_start=1 /gene="pelB (fragment)" /product="leader peptide for secretion" /label=pelB signal sequence /translation="MKYLLPTAAAGLLLLAAQPAMA" CDS 485..502 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" terminator 569..616 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 653..1108 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" CDS complement(1200..2015) /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 2137..2725 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature 2911..3050 /label=bom /note="basis of mobility region from pBR322" CDS complement(3152..3343) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4152..5234) /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(5235..5312) /gene="lacI" /label=lacI promoter ORIGIN 1 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag 61 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa 121 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag 181 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg 241 atcgagatct cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata 301 acaattcccc tctagaaata attttgttta actttaagaa ggagatatac atatgaaata 361 cctgctgccg accgctgctg ctggtctgct gctcctcgct gcccagccgg cgatggccat 421 ggatatcgga attaattcgg atccgaattc gagctccgtc gacaagcttg cggccgcact 481 cgagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga 541 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt 601 cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc 661 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt 721 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc 781 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta 841 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc 901 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg 961 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt 1021 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat 1081 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa 1141 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct 1201 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata 1261 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat 1321 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct 1381 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact 1441 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag 1501 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc 1561 gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa 1621 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat 1681 tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca 1741 tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt 1801 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac 1861 aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca 1921 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc 1981 ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg 2041 taagcagaca gttttattgt tcatgaccaa aatcccttaa cgtgagtttt cgttccactg 2101 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt 2161 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca 2221 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac 2281 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac 2341 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct 2401 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg 2461 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca 2521 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt 2581 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta 2641 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc 2701 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc 2761 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa 2821 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag 2881 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct 2941 gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag 3001 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc 3061 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac 3121 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac 3181 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg 3241 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga 3301 taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag 3361 ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac 3421 gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg 3481 tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata 3541 cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa 3601 tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca 3661 ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc 3721 gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca 3781 acgacaggag cacgatcatg cgcacccgtg gggccgccat gccggcgata atggcctgct 3841 tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga 3901 ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc 3961 cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag 4021 tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga 4081 aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa 4141 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 4201 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt 4261 ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc 4321 agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac 4381 ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca 4441 ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg 4501 gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa 4561 ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg 4621 agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct 4681 aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg 4741 tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac 4801 gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag 4861 ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct 4921 tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga 4981 gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg 5041 ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc 5101 agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg 5161 ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac 5221 gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata 5281 ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga 5341 ctcctgcatt aggaag //