LOCUS Exported 4843 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBbB6k-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4843) AUTHORS Lee TS, Krupa RA, Zhang F, Hajimorad M, Holtz WJ, Prasad N, Lee SK, Keasling JD TITLE BglBrick vectors and datasheets: A synthetic biology platform for gene expression. JOURNAL J Biol Eng. 2011 Sep 20;5:12. PUBMED 21933410 REFERENCE 2 (bases 1 to 4843) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4843 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(49..1131) /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 1093..1112 /label=LacI-R /note="LacI, reverse primer" promoter complement(1132..1209) /gene="lacI" /label=lacI promoter primer_bind 1415..1434 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" protein_bind 1502..1518 /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)." protein_bind 1525..1541 /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 1609..2325 /codon_start=1 /product="GFP variant optimized for excitation by UV light" /label=GFPuv /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" primer_bind complement(1639..1667) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 2262..2282 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" terminator 2358..2429 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2445..2472 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(2539..3201) /codon_start=1 /product="replication protein for the broad-host-range plasmid pBBR1 from Bordetella bronchiseptica" /label=pBBR1 Rep /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVLAYVV NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR" terminator 3788..3882 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(3913..4707) /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4024..4043) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 4634..4653 /label=Neo-R /note="Neomycin resistance gene, reverse primer" ORIGIN 1 gacgtcggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct 61 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 121 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag 181 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg 241 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc 301 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt 361 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac 421 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc 481 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag 541 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc 601 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact 661 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc 721 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg 781 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat 841 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg 901 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc 961 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac 1021 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg 1081 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac 1141 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc 1201 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca 1261 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg 1321 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca 1381 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag 1441 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga 1501 attgtgagcg gataacaatt gacattgtga gcggataaca agatactgag cacatcagca 1561 ggacgcactg accgaattca aaagatcttt taagaaggag atatacatat gagtaaagga 1621 gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg 1681 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt 1741 aaatttattt gcactactgg aaaactacct gttccgtggc caacacttgt cactactttc 1801 tcttatggtg ttcaatgctt ttcccgttat ccggatcaca tgaaacggca tgactttttc 1861 aagagtgcca tgcccgaagg ttatgtacag gaacgcacta tatctttcaa agatgacggg 1921 aactacaaga cgcgtgctga agtcaagttt gaaggtgata cccttgttaa tcgtatcgag 1981 ttaaaaggta ttgattttaa agaagatgga aacattctcg gacacaaact ggagtacaac 2041 tataactcac acaatgtata catcacggca gacaaacaaa agaatggaat caaagctaac 2101 ttcaaaattc gccacaacat tgaagatggc tccgttcaac tagcagacca ttatcaacaa 2161 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa 2221 tctgcccttt cgaaagatcc caacgaaaag cgtgaccaca tggtccttct tgagtttgta 2281 actgctgctg ggattacaca tggcatggat gagctctaca aataaggatc caaactcgag 2341 taaggatctc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt 2401 ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg 2461 ggcctttctg cgtttatacc taggctacag ccgatagtct ggaacagcgc acttacgggt 2521 tgctgcgcaa cccaagtgct accggcgcgg cagcgtgacc cgtgtcggcg gctccaacgg 2581 ctcgccatcg tccagaaaac acggctcatc gggcatcggc aggcgctgct gcccgcgccg 2641 ttcccattcc tccgtttcgg tcaaggctgg caggtctggt tccatgcccg gaatgccggg 2701 ctggctgggc ggctcctcgc cggggccggt cggtagttgc tgctcgcccg gatacagggt 2761 cgggatgcgg cgcaggtcgc catgccccaa cagcgattcg tcctggtcgt cgtgatcaac 2821 caccacggcg gcactgaaca ccgacaggcg caactggtcg cggggctggc cccacgccac 2881 gcggtcattg accacgtagg ccaacacggt gccggggccg ttgagcttca cgacggagat 2941 ccagcgctcg gccaccaagt ccttgactgc gtattggacc gtccgcaaag aacgtccgat 3001 gagcttggaa agtgtcttct ggctgaccac cacggcgttc tggtggccca tctgcgccac 3061 gaggtgatgc agcagcattg ccgccgtggg tttcctcgca ataagcccgg cccacgcctc 3121 atgcgctttg cgttccgttt gcacccagtg accgggcttg ttcttggctt gaatgccgat 3181 ttctctggac tgcgtggcca tgcttatctc catgcggtag gggtgccgca cggttgcggc 3241 accatgcgca atcagctgca acttttcggc agcgcgacaa caattatgcg ttgcgtaaaa 3301 gtggcagtca attacagatt ttctttaacc tacgcaatga gctattgcgg ggggtgccgc 3361 aatgagctgt tgcgtacccc ccttttttaa gttgttgatt tttaagtctt tcgcatttcg 3421 ccctatatct agttctttgg tgcccaaaga agggcacccc tgcggggttc ccccacgcct 3481 tcggcgcggc tccccctccg gcaaaaagtg gcccctccgg ggcttgttga tcgactgcgc 3541 ggccttcggc cttgcccaag gtggcgctgc ccccttggaa cccccgcact cgccgccgtg 3601 aggctcgggg ggcaggcggg cgggcttcgc ccttcgactg cccccactcg cataggcttg 3661 ggtcgttcca ggcgcgtcaa ggccaagccg ctgcgcggtc gctgcgcgag ccttgacccg 3721 ccttccactt ggtgtccaac cggcaagcga agcgcgcagg ccgcaggccg gaggcactag 3781 tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca 3841 gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag ctctcgaacc 3901 ccagagtccc gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc 3961 gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc 4021 agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc 4081 acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc 4141 gccatgggtc acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag 4201 ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc 4261 ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt 4321 agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc 4381 aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc 4441 ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag 4501 ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt 4561 gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc 4621 gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc 4681 tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcaga 4741 tcatgatccc ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca 4801 gggcttccca accttaccag agggcgcccc agctggcaat tcc //