LOCUS Exported 6186 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 6186) AUTHORS 11111111 TITLE Direct Submission JOURNAL Exported Sep 10, 2025 from SnapGene 6.0.2 https://www.snapgene.com FEATURES Location/Qualifiers source 1..6186 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 127..145 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 146..170 /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 215..922 /codon_start=1 /gene="dsbC" /product="bacterial periplasmic protein disulfide isomerase" /label=DsbC /translation="MKKGFMLFTLLAAFSGFAQADDAAIQQTLAKMGIKSSDIQPAPVA GMKTVLTNSGVLYITDDGKHIIQGPMYDVSGTAPVNVTNKMLLKQLNALEKEMIVYKAP QEKHVITVFTDITCGYCHKLHEQMADYNALGITVRYLAFPRQGLDSDAEKEMKAIWCAK DKNKAFDDVMAGKSVAPASCDVDIADHYALGVQLGVSGTPAVVLSNGTLVPGYQPPKEM KEFLDEHQKMTSGK" CDS 944..961 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 971..988 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 1004..1048 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 1064..1078 /codon_start=1 /product="enterokinase recognition and cleavage site" /label=enterokinase site /translation="DDDDK" CDS 1157..1180 /codon_start=1 /product="8xHis affinity tag" /label=8xHis /translation="HHHHHHHH" terminator 1257..1304 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 1341..1796 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" CDS complement(1888..2703) /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 2825..3413 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature 3599..3738 /label=bom /note="basis of mobility region from pBR322" CDS complement(3840..4031) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4840..5922) /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(5923..6000) /gene="lacI" /label=lacI promoter ORIGIN 1 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg 61 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg 121 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa 181 taattttgtt taactttaag aaggagatat acatatgaag aaaggtttta tgttgtttac 241 tttgttagcg gcgttttcag gctttgctca ggctgatgac gcggcaattc aacaaacgtt 301 agccaaaatg ggcatcaaaa gcagcgatat tcagcccgcg cctgtagctg gcatgaagac 361 agttctgact aacagcggcg tgttgtacat caccgatgat ggtaaacata tcattcaggg 421 gccaatgtat gacgttagtg gcacggctcc ggtcaatgtc accaataaga tgctgttaaa 481 gcagttgaat gcgcttgaaa aagagatgat cgtttataaa gcgccgcagg aaaaacacgt 541 catcaccgtg tttactgata ttacctgtgg ttactgccac aaactgcatg agcaaatggc 601 agactacaac gcgctgggga tcaccgtgcg ttatcttgct ttcccgcgcc aggggctgga 661 cagcgatgca gagaaagaaa tgaaagctat ctggtgtgcg aaagataaaa acaaagcgtt 721 tgatgatgtg atggcaggta aaagcgtcgc accagccagt tgcgacgtgg atattgccga 781 ccattacgca cttggcgtcc agcttggcgt tagcggtact ccggcagttg tgctgagcaa 841 tggcacactt gttccgggtt accagccgcc gaaagagatg aaagaatttc tcgacgaaca 901 ccaaaaaatg accagcggta aaggatcaac tagtggttct ggtcatcacc atcaccatca 961 ctccgcgggt ctggtgccac gcggtagtac tgcaattggt atgaaagaaa ccgctgctgc 1021 taaattcgaa cgccagcaca tggacagccc agatctgggt accgatgacg acgacaagag 1081 cccgggcttc tcctcaacca tggcgatatc ggatccgaat tcgagctccg tcgacaagct 1141 tgcggccgca ctcgagcacc accaccacca ccaccaccac taattgatta atacctaggc 1201 tgctaaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag 1261 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta 1321 tatccggatt ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt 1381 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt 1441 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct 1501 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg 1561 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga 1621 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc 1681 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga 1741 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt 1801 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca 1861 aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg 1921 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga 1981 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat 2041 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc 2101 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat 2161 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc 2221 aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt 2281 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc 2341 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg 2401 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg 2461 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc 2521 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg 2581 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc 2641 agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct 2701 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgaccaaaa 2761 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat 2821 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc 2881 taccagcggt ggtttgtttg ccggatcaag agctacaaac tctttttccg aaggtaactg 2941 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc 3001 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg 3061 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg 3121 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa 3181 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg 3241 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga 3301 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct 3361 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca 3421 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc 3481 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg 3541 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc 3601 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct 3661 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt 3721 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct 3781 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt 3841 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg 3901 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc 3961 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc 4021 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg 4081 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg 4141 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact 4201 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag 4261 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga 4321 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg 4381 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg 4441 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggg 4501 gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg 4561 acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc 4621 gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt 4681 cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc 4741 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt 4801 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg 4861 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg 4921 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc 4981 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag 5041 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc 5101 gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg 5161 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc 5221 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc 5281 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg 5341 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac 5401 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg 5461 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc 5521 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag 5581 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac 5641 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc 5701 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg 5761 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg 5821 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac 5881 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg 5941 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc 6001 cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt 6061 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca 6121 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga 6181 agtggc //