LOCUS Exported 6147 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION csd1, construct: 54-312, pET21a(+) C-tag. ACCESSION . VERSION . KEYWORDS csd1_54-312/pET21a(+) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6147) TITLE Structural basis of the heterodimer formation between cell shape-determining proteins Csd1 and Csd2 from Helicobacter pylori JOURNAL Unpublished REFERENCE 2 (bases 1 to 6147) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6147 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 83..671 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 572..591 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 825..842 /label=L4440 /note="L4440 vector, forward primer" misc_feature 857..999 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(925..944) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1085..1107 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(1101..1292) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(2101..3183) /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 3145..3164 /label=LacI-R /note="LacI, reverse primer" promoter complement(3184..3261) /gene="lacI" /label=lacI promoter primer_bind 3467..3486 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 3570..3589 /label=T7 /note="T7 promoter, forward primer" promoter 3570..3588 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 3589..3613 /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 3628..3650 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 4444..4461 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(4514..4532) /label=T7 Term /note="T7 terminator, reverse primer" terminator 4528..4575 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 4612..5067 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4699..4718) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4909..4930 /label=F1ori-F /note="F1 origin, forward primer" promoter 5094..5198 /gene="bla" /label=AmpR promoter CDS 5199..6059 /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 complement(5417..5436) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 61 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 121 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 181 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 241 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 301 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 361 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 421 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 481 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 541 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 601 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 661 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 721 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 781 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 841 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac 901 accgcatata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta 961 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc 1021 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc 1081 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag 1141 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc 1201 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc 1261 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg 1321 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat 1381 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg 1441 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt 1501 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc 1561 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt 1621 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat 1681 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc 1741 acgatcatgc gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa 1801 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc 1861 gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc 1921 cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg 1981 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag 2041 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc 2101 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 2161 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga 2221 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc 2281 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata 2341 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag 2401 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat 2461 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc 2521 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg 2581 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat 2641 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga 2701 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt 2761 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag 2821 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct 2881 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc 2941 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa 3001 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat 3061 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg 3121 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt 3181 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt 3241 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta 3301 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat 3361 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga 3421 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga 3481 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt 3541 agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc 3601 ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg 3661 gagattaaaa acattgacaa gcagcatgct ctaatcacta aagaatttga gaaaaaaaaa 3721 gagacgaatg aaaagctttc cttgcaaatg gatgaatttt tagacgattt gcagctttca 3781 ggggaacgca tcaacgattt agaagaagtg gtgggagtga ataggcctga agaagaaaaa 3841 gaagagggca atttttccag tcgtttggat gtcgctggga ttaccgggct tcaaaaaagc 3901 tttatcatgc gccttatccc taatgactac ccgctagaat cctatcggcg cgtttcagcc 3961 gcctttaata aaagaatcca ccctattttg catgtgctgc acaaccatac cgggcttgat 4021 ttaagcaccg ctattaacac ccctgtgtat gcgagcgcga gcggggtagt agggttagcg 4081 agcaaggggt ggaatggggg gtatgggaat ttgattaaag ttttccaccc ttttggtttt 4141 aaaacctact acgcccattt gaataaaatc gtcgtaaaaa cgggcgaatt tgtcaaaaaa 4201 gggcagttga ttgggtatag tggtaataca gggatgagca cagggccgca tttgcattat 4261 gaagtgcgtt ttttagatca acccataaac cccatgagtt tcaccaaatg gaacatgaaa 4321 gattttgaag aagttttcaa taaagaaagg agcatcagat ggcaatcttt gataacaata 4381 ataaatcggc taatgcaaaa acaggaccag cgactatcat cgctcaaggc acaaaaactc 4441 gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa ggaagctgag 4501 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc 4561 ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg gacgcgccct 4621 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg 4681 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg 4741 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac 4801 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct 4861 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt 4921 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt 4981 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt 5041 ttaacaaaat attaacgttt acaatttcag gtggcacttt tcggggaaat gtgcgcggaa 5101 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 5161 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg 5221 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc 5281 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg 5341 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga 5401 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc 5461 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag 5521 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga 5581 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg 5641 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga 5701 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt 5761 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact 5821 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt 5881 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg 5941 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta 6001 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac 6061 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta 6121 aaaggatcta ggtgaagatc ctttttg //