LOCUS Exported 6180 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pET-21a(+)-LpxA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6180) AUTHORS Lee BI, Suh SW TITLE Crystal structure of UDP-N-acetylglucosamine acyltransferase from Helicobacter pylori. JOURNAL Proteins. 2003 Nov 15. 53(3):772-4. PUBMED 14579368 REFERENCE 2 (bases 1 to 6180) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6180 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 26..73 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind 69..87 /label=T7 Term /note="T7 terminator, reverse primer" CDS complement(140..157) /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" RBS 984..1006 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" protein_bind 1021..1045 /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)." primer_bind complement(1045..1064) /label=T7 /note="T7 promoter, forward primer" promoter complement(1046..1064) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1148..1167) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" promoter 1373..1450 /gene="lacI" /label=lacI promoter CDS 1451..2533 /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" primer_bind complement(1470..1489) /label=LacI-R /note="LacI, reverse primer" CDS 3342..3533 /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" primer_bind complement(3527..3549) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 3635..3777 /label=bom /note="basis of mobility region from pBR322" primer_bind 3690..3709 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(3792..3809) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3963..4551) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4043..4062) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4722..5582) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 5345..5364 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5583..5687) /gene="bla" /label=AmpR promoter rep_origin complement(5714..6169) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5851..5872) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6063..6082 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa 61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt 121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagttcctct tgtttttctt 181 cgctagaata ttcgcttgac ttatacgcca cgcccctaga actctctaaa ataaaagagc 241 aaatctcttt cacaaaaggg ttattggcat gctcttctaa ttctaattta gcgctctctc 301 ttaaagaggg gatagggcgg aacaatcttt tataaagtgc ataaataaaa tcaatatcct 361 tactctctaa taattggcgc atccggtggc ggtttaaccc cctaataaaa gcgcgattgc 421 cctctacggt gcaataaggc ggcacgtctt tgcctaaagc gctcttaccg gctatcatgc 481 accctttagc gatacgcaca aactgatgga tcgcggtaag accgccgata ttgacataat 541 cgcctatttc aatatgacct gctaaagtta cgccattagc caaaatacaa tggcttccaa 601 tcacgcaatc atgagcgaca tgcacataag ccatgagcag gtttttatcc ccaataaggg 661 tttttttaat ccccccttca gtgccgggat ttatcatgca aaattcccga ataaggttgt 721 cttctccaat aatcagttcg ctgtattcgc ccttatattt taaatcttga ggctgtgtgc 781 ctagcacggc aaaagggaaa atttcggtgt ttttcccaac gaaagtatgc ccttgtaagg 841 ttacattgtt gtggagtttc acgccttcat ctagtttgac gccatctcca atcacgcaaa 901 attccccaat ctctacgccc ttattaatct ctgctttagg agagatgatg gctgtttttg 961 caatcttact catcatatgt atatctcctt cttaaagtta aacaaaatta tttctagagg 1021 ggaattgtta tccgctcaca attcccctat agtgagtcgt attaatttcg cgggatcgag 1081 atctcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt 1141 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc 1201 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc 1261 gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta 1321 ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat 1381 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag 1441 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta 1501 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa 1561 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc 1621 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc 1681 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc 1741 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca 1801 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga 1861 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa 1921 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt 1981 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg 2041 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga 2101 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat 2161 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat 2221 taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga 2281 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg 2341 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca 2401 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc 2461 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga 2521 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcaccgggat 2581 ctcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca 2641 tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc 2701 cggcagcgct ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg 2761 gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc 2821 ccgccaccaa acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccccacggg 2881 tgcgcatgat cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg 2941 gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt 3001 ctgcgacctg agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa 3061 cgcggaagtc agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc 3121 taccctgtgg aacacctaca tctgtattaa cgaagcgctg gcattgaccc tgagtgattt 3181 ttctctggtc ccgccgcatc cataccgcca gttgtttacc ctcacaacgt tccagtaacc 3241 gggcatgttc atcatcagta acccgtatcg tgagcatcct ctctcgtttc atcggtatca 3301 ttacccccat gaacagaaat cccccttaca cggaggcatc agtgaccaaa caggaaaaaa 3361 ccgcccttaa catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca 3421 acgagctgga cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg 3481 agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc 3541 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc 3601 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg 3661 atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca 3721 ccatatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc 3781 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 3841 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 3901 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 3961 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 4021 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 4081 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 4141 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 4201 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 4261 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 4321 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 4381 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt 4441 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 4501 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 4561 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 4621 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 4681 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 4741 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 4801 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 4861 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 4921 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 4981 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca 5041 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 5101 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 5161 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 5221 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 5281 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 5341 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 5401 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 5461 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 5521 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 5581 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 5641 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 5701 aaagtgccac ctgaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt 5761 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa 5821 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag 5881 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt 5941 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac 6001 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag 6061 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg 6121 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca //