LOCUS Exported 8656 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS EC.SurA.pTYB1.a21-428.wt SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8656) AUTHORS Bitto E, McKay DB TITLE Crystallographic structure of SurA, a molecular chaperone that facilitates folding of outer membrane porins. JOURNAL Structure. 2002 Nov . 10(11):1489-98. PUBMED 12429090 REFERENCE 2 (bases 1 to 8656) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8656 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 35..139 /gene="bla" /label=AmpR promoter CDS 140..1000 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(358..377) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin complement(1042..1555) /direction=LEFT /label=M13 ori /note="M13 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1237..1258) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1449..1468 /label=F1ori-R /note="F1 origin, reverse primer" rep_origin 1666..2254 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2155..2174 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2408..2425 /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(2447..2466) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2607..2629 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(2623..2814) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(3371..4453) /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 4415..4434 /label=LacI-R /note="LacI, reverse primer" promoter complement(4454..4531) /gene="lacI" /label=lacI promoter terminator 4684..4770 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 4867..4953 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 5050..5136 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 5233..5319 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 5416..5502 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 5637..5656 /label=T7 /note="T7 promoter, forward primer" promoter 5637..5655 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 5656..5680 /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 5695..5717 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 6955..7773 /codon_start=1 /gene="Saccharomyces cerevisiae VMA1" /product="5' region of the intein from the yeast Vma1 subunit of the vacuolar ATPase (Chong et al., 1998)" /label=Sce VMA intein 5' region /translation="CFAKGTNVLMADGSIECIENIEVGNKVMGKDGRPREVIKLPRGRE TMYSVVQKSQHRAHKSDSSREVPELLKFTCNATHELVVRTPRSVRRLSRTIKGVEYFEV ITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELVESYRKASNKAYFEWTIEARD LSLLGSHVRKATYQTYAPILYENDHFFDYMQKSKFHLTIEGPKVLAYLLGLWIGDGLSD RATFSVDSRDTSLMERVTEYAEKLNLCAEYKDRKEPQVAKTVNLYSKVVRG" CDS 8229..8240 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" CDS 8350..8505 /codon_start=1 /gene="Bacillus circulans chiA" /product="chitin binding domain from chitinase A1 (Watanabe et al., 1994)" /label=CBD /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV PALWQLQ" primer_bind complement(8570..8588) /label=T7 Term /note="T7 terminator, reverse primer" terminator 8584..8631 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" ORIGIN 1 aactacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt 61 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata 121 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt 181 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc 241 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat 301 ccttgagagt tttcgccccg aagaacgttc tccaatgatg agcactttta aagttctgct 361 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca 421 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg 481 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa 541 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg 601 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga 661 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg 721 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt 781 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg 841 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc 901 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca 961 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc 1021 atatatactt tagattgatt taccccggtt gataatcaga aaagccccaa aaacaggaag 1081 attgtataag caaatattta aattgtaaac gttaatattt tgttaaaatt cgcgttaaat 1141 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa 1201 tcaaaagaat agcccgagat agggttgagt gttgttccag tttggaacaa gagtccacta 1261 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca 1321 ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa agcactaaat 1381 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg 1441 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc 1501 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtaaaag 1561 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc 1621 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt 1681 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt 1741 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat 1801 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc 1861 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa 1921 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg 1981 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag 2041 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag 2101 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa 2161 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt 2221 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg 2281 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc 2341 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac 2401 cgagcgcagc gagtcagtga gcgaggaagc tatggtgcac tctcagtaca atctgctctg 2461 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc 2521 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc 2581 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc 2641 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gcagcgattc 2701 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt 2761 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc 2821 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat 2881 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa 2941 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcc 3001 gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt 3061 ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat 3121 tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc 3181 gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt 3241 cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa 3301 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat 3361 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 3421 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt 3481 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca 3541 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg 3601 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac 3661 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg 3721 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac 3781 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga 3841 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta 3901 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt 3961 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg 4021 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt 4081 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt 4141 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag 4201 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc 4261 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca 4321 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt 4381 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg 4441 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac 4501 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac 4561 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg 4621 aatggtgcat gccggcatgc cgccctttcg tcttcaagaa ttaattccca attccccagg 4681 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg 4741 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag 4801 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggaa ttaattcccc 4861 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt 4921 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg 4981 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gaattaattc 5041 cccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt 5101 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt 5161 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggaattaa 5221 ttccccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 5281 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 5341 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggaat 5401 taattcccca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt 5461 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg 5521 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg 5581 aattggggat cggaattaat tcccggttta aaccggggat ctcgatcccg cgaaattaat 5641 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt 5701 taactttaag aaggagatat acatatggcc ccccaggtag tcgataaagt cgcagccgtc 5761 gtcaataacg gcgtcgtgct ggaaagcgac gttgatggat taatgcagtc ggtaaaactg 5821 aacgctgctc aggcaaggca gcaacttcct gatgacgcga cgctgcgcca ccaaatcatg 5881 gaacgtttga tcatggatca aatcatcctg cagatggggc agaaaatggg agtgaaaatc 5941 tccgatgagc agctggatca ggcgattgct aacatcgcga aacagaacaa catgacgctg 6001 gatcagatgc gcagccgtct ggcttacgat ggactgaact acaacaccta tcgtaaccag 6061 atccgcaaag agatgattat ctctgaagtg cgtaacaacg aggtgcgtcg tcgcatcacc 6121 atcctgccgc aggaagtcga atccctggcg cagcaggtgg gtaaccaaaa cgacgccagc 6181 actgagctga acctgagcca catcctgatc ccgctgccgg aaaacccgac ctctgatcag 6241 gtgaacgaag cggaaagcca ggcgcgcgcc attgtcgatc aggcgcgtaa cggcgctgat 6301 ttcggtaagc tggcgattgc tcattctgcc gaccagcagg cgctgaacgg cggccagatg 6361 ggctggggcc gtattcagga gttgcccggg atcttcgccc aggcattaag caccgcgaag 6421 aaaggcgaca ttgttggccc gattcgttcc ggcgttggct tccatattct gaaagttaac 6481 gacctgcgcg gcgaaagcaa aaatatctcg gtgaccgaag ttcatgctcg ccatattctg 6541 ctgaaaccgt cgccgatcat gactgacgaa caggcccgtg tgaaactgga acagattgct 6601 gctgatatca agagtggtaa aacgactttt gctgccgcag cgaaagagtt ctctcaggat 6661 ccaggctctg ctaaccaggg cggcgatctc ggctgggcta caccagatat tttcgatccg 6721 gccttccgtg acgccctgac tcgcctgaac aaaggtcaaa tgagtgcacc ggttcactct 6781 tcattcggct ggcatttaat cgaactgctg gatacccgta atgtcgataa aaccgacgct 6841 gcgcagaaag atcgtgcata ccgcatgctg atgaaccgta agttctcgga agaagcagca 6901 agctggatgc aggaacaacg tgccagcgcc tacgttaaaa tcctgagcaa cgggtgcttt 6961 gccaagggta ccaatgtttt aatggcggat gggtctattg aatgtattga aaacattgag 7021 gttggtaata aggtcatggg taaagatggc agacctcgtg aggtaattaa attgcccaga 7081 ggaagagaaa ctatgtacag cgtcgtgcag aaaagtcagc acagagccca caaaagtgac 7141 tcaagtcgtg aagtgccaga attactcaag tttacgtgta atgcgaccca tgagttggtt 7201 gttagaacac ctcgtagtgt ccgccgtttg tctcgtacca ttaagggtgt cgaatatttt 7261 gaagttatta cttttgagat gggccaaaag aaagcccccg acggtagaat tgttgagctt 7321 gtcaaggaag tttcaaagag ctacccaata tctgaggggc ctgagagagc caacgaatta 7381 gtagaatcct atagaaaggc ttcaaataaa gcttattttg agtggactat tgaggccaga 7441 gatctttctc tgttgggttc ccatgttcgt aaagctacct accagactta cgctccaatt 7501 ctttatgaga atgaccactt tttcgactac atgcaaaaaa gtaagtttca tctcaccatt 7561 gaaggtccaa aagtacttgc ttatttactt ggtttatgga ttggtgatgg attgtctgac 7621 agggcaactt tttcggttga ttccagagat acttctttga tggaacgtgt tactgaatat 7681 gctgaaaagt tgaatttgtg cgccgagtat aaggacagaa aagaaccaca agttgccaaa 7741 actgttaatt tgtactctaa agttgtcaga ggtaatggta ttcgcaataa tcttaatact 7801 gagaatccat tatgggacgc tattgttggc ttaggattct tgaaggacgg tgtcaaaaat 7861 attccttctt tcttgtctac ggacaatatc ggtactcgtg aaacatttct tgctggtcta 7921 attgattctg atggctatgt tactgatgag catggtatta aagcaacaat aaagacaatt 7981 catacttctg tcagagatgg tttggtttcc cttgctcgtt ctttaggctt agtagtctcg 8041 gttaacgcag aacctgctaa ggttgacatg aatgtcacca aacataaaat tagttatgct 8101 atttatatgt ctggtggaga tgttttgctt aacgttcttt cgaagtgtgc cggctctaaa 8161 aaattcaggc ctgctcccgc cgctgctttt gcacgtgagt gccgcggatt ttatttcgag 8221 ttacaagaat tgaaggaaga cgattattat gggattactt tatctgatga ttctgatcat 8281 cagtttttgc ttggatccca ggttgtcgtc catgcatgcg gtggcctgac cggtctgaac 8341 tcaggcctca cgacaaatcc tggtgtatcc gcttggcagg tcaacacagc ttatactgcg 8401 ggacaattgg tcacatataa cggcaagacg tataaatgtt tgcagcccca cacctccttg 8461 gcaggatggg aaccatccaa cgttcctgcc ttgtggcagc ttcaatgact gcaggaaggg 8521 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa 8581 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga 8641 ggaactatat ccggat //