LOCUS Exported 5368 bp DNA circular SYN 19-MAR-1980 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS pET-28b(+). SOURCE synthetic DNA construct. ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5368) TITLE Direct Submission REFERENCE 2 (bases 1 to 5368) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 13, 2025 from SnapGene 4.1.9 http://www.snapgene.com COMMENT This file is created by Vector NTI http://www.invitrogen.com/ COMMENT ORIGDB|GenBank COMMENT LSOWNER| COMMENT VNTNAME|Exported| COMMENT Vector_NTI_Display_Data_(Do_Not_Edit!) 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join(485..5368,1..484) /organism="synthetic DNA construct" /mol_type="other DNA" /vntifkey="98" promoter 100..118 /vntifkey="30" /label=T7\promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 119..143 /bound_moiety="lac repressor encoded by lacI" /vntifkey="31" /label=lac\operator /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 158..180 /vntifkey="32" /label=RBS_1 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 199..216 /codon_start=1 /product="6xHis affinity tag" /vntifkey="4" /label=6xHis CDS 226..243 /codon_start=1 /product="thrombin recognition and cleavage site" /vntifkey="4" /label=thrombin\site CDS 247..279 /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /vntifkey="4" /label=T7\tag\(gene\10\leader) /note="promotes efficient translation in E. coli" CDS 328..345 /codon_start=1 /product="6xHis affinity tag" /vntifkey="4" /label=6xHis terminator 412..459 /vntifkey="43" /label=T7\terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 496..951 /direction=RIGHT /vntifkey="33" /label=f1\ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" CDS complement(1044..1859) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /vntifkey="4" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" rep_origin 1981..2569 /direction=RIGHT /vntifkey="33" /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature 2755..2897 /vntifkey="21" /label=bom /note="basis of mobility region from pBR322" CDS complement(2999..3190) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /vntifkey="4" /label=rop CDS complement(3999..5081) /codon_start=1 /gene="lacI" /product="lac repressor" /vntifkey="4" /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)." promoter 5082..5159 /gene="lacI" /vntifkey="30" /label=lacI\promoter BASE COUNT 1266 a 1443 c 1391 g 1268 t ORIGIN 1 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg 61 cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg 121 aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga 181 tataccatgg gcagcagcca tcatcatcat catcacagca gcggcctggt gccgcgcggc 241 agccatatgg ctagcatgac tggtggacag caaatgggtc gggatccgaa ttcgagctcc 301 gtcgacaagc ttgcggccgc actcgagcac caccaccacc accactgaga tccggctgct 361 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa 421 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc 481 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta 541 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc 601 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt 661 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg 721 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca 781 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct 841 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga 901 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt tcaggtggca 961 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata 1021 tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg aaactgcaat 1081 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga 1141 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg 1201 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt 1261 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct 1321 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc 1381 aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa 1441 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca 1501 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc 1561 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga 1621 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg 1681 ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag 1741 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca 1801 tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata 1861 acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga ccaaaatccc 1921 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 1981 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 2041 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 2101 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt 2161 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 2221 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 2281 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 2341 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 2401 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 2461 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 2521 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 2581 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 2641 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg 2701 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat 2761 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatatg gtgcactctc 2821 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg 2881 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt 2941 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc 3001 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt 3061 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct 3121 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct 3181 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga 3241 tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg 3301 aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc 3361 agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc 3421 atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac 3481 tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc 3541 agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc 3601 aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg 3661 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga 3721 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg 3781 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc 3841 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg 3901 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg 3961 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg 4021 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 4081 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc 4141 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg 4201 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg 4261 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc 4321 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca 4381 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc 4441 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc 4501 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc 4561 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt 4621 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca 4681 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga 4741 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc 4801 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg 4861 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt 4921 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc 4981 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca 5041 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga 5101 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc 5161 gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt 5221 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag 5281 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa 5341 gtggcgagcc cgatcttccc catcggtg //