LOCUS Exported 7718 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION lacritin-intein fusion plasmid coding for truncated lacritin without signal peptide. ACCESSION . VERSION . KEYWORDS C25-pLAC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7718) AUTHORS Wang J, Wang N, Xie J, Walton SC, McKown RL, Raab RW, Ma P, Beck SL, Coffman GL, Hussaini IM, Laurie GW TITLE Restricted epithelial proliferation by lacritin via PKCalpha-dependent NFAT and mTOR pathways. JOURNAL J Cell Biol. 2006 Aug 28;174(5):689-700. Epub 2006 Aug 21. PUBMED 16923831 REFERENCE 2 (bases 1 to 7718) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7718 /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(3688..4770) /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 4732..4751 /label=LacI-R /note="LacI, reverse primer" promoter complement(4771..4848) /gene="lacI" /label=lacI promoter terminator 5001..5087 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 5184..5270 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 5367..5453 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 5550..5636 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" CDS 6017..6835 /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 7291..7302 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" CDS 7412..7567 /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(7632..7650) /label=T7 Term /note="T7 terminator, reverse primer" terminator 7646..7693 /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 ccgatggaag atgcctcctc tgactcgacg ggtgctgatc 2761 ctgcccagga agctgggacc tctaagccta atgaagagat ctcaggtcca gcagaaccag 2821 cttcaccccc agagacaacc acaacagccc aggagacttc ggcggcagca gttcagggga 2881 cagccaaggt cacctcaagc aggcaggaac taaaccccct gaaatccata gtggagaaaa 2941 gtatcttact aacagaacaa gcccttgcaa aagcaggaaa aggaatgcac ggaggcgtgc 3001 caggtggaaa acaatccatc gaaaatggaa gtgaatttgc acaaaaatta ctgaagaaat 3061 tcagtctatt aaaaccatgg gcatgagata aagcgggcca tgttaagggc ggttttttcc 3121 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg 3181 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg 3241 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact 3301 cagggtcaat gccagccgaa cgccagcaag acgtagccca gcgcgtcggc cgccatgccg 3361 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga 3421 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag 3481 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc 3541 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag 3601 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg 3661 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 3721 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 3781 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg 3841 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg 3901 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac 3961 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag 4021 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg 4081 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg 4141 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga 4201 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac 4261 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga 4321 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg 4381 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac 4441 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc 4501 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag 4561 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg 4621 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga 4681 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc 4741 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg 4801 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac 4861 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga 4921 gcaccgccgc cgcaaggaat ggtgcatgcc ggcatgccgc cctttcgtct tcaagaatta 4981 attcccaatt ccccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc 5041 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg 5101 atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg 5161 ccaggaatta attccccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct 5221 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag 5281 cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa 5341 ctgccaggaa ttaattcccc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg 5401 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg 5461 gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat 5521 aaactgccag gaattaattc cccaggcatc aaataaaacg aaaggctcag tcgaaagact 5581 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc 5641 cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc 5701 cataaactgc caggaattaa ttcatggaag atgcctcctc tgactcgacg ggtgctgatc 5761 ctgcccagga agctgggacc tctaagccta atgaagagat ctcaggtcca gcagaaccag 5821 cttcaccccc agagacaacc acaacagccc aggagacttc ggcggcagca gttcagggga 5881 cagccaaggt cacctcaagc aggcaggaac taaaccccct gaaatccata gtggagaaaa 5941 gtatcttact aacagaacaa gcccttgcaa aagcaggaaa aggaatgcac ggaggcgtgc 6001 caggtggatg acttcctgct ttgccaaggg taccaatgtt ttaatggcgg atgggtctat 6061 tgaatgtatt gaaaacattg aggttggtaa taaggtcatg ggtaaagatg gcagacctcg 6121 tgaggtaatt aaattgccca gaggaagaga aactatgtac agcgtcgtgc agaaaagtca 6181 gcacagagcc cacaaaagtg actcaagtcg tgaagtgcca gaattactca agtttacgtg 6241 taatgcgacc catgagttgg ttgttagaac acctcgtagt gtccgccgtt tgtctcgtac 6301 cattaagggt gtcgaatatt ttgaagttat tacttttgag atgggccaaa agaaagcccc 6361 cgacggtaga attgttgagc ttgtcaagga agtttcaaag agctacccaa tatctgaggg 6421 gcctgagaga gccaacgaat tagtagaatc ctatagaaag gcttcaaata aagcttattt 6481 tgagtggact attgaggcca gagatctttc tctgttgggt tcccatgttc gtaaagctac 6541 ctaccagact tacgctccaa ttctttatga gaatgaccac tttttcgact acatgcaaaa 6601 aagtaagttt catctcacca ttgaaggtcc aaaagtactt gcttatttac ttggtttatg 6661 gattggtgat ggattgtctg acagggcaac tttttcggtt gattccagag atacttcttt 6721 gatggaacgt gttactgaat atgctgaaaa gttgaatttg tgcgccgagt ataaggacag 6781 aaaagaacca caagttgcca aaactgttaa tttgtactct aaagttgtca gaggtaatgg 6841 tattcgcaat aatcttaata ctgagaatcc attatgggac gctattgttg gcttaggatt 6901 cttgaaggac ggtgtcaaaa atattccttc tttcttgtct acggacaata tcggtactcg 6961 tgaaacattt cttgctggtc taattgattc tgatggctat gttactgatg agcatggtat 7021 taaagcaaca ataaagacaa ttcatacttc tgtcagagat ggtttggttt cccttgctcg 7081 ttctttaggc ttagtagtct cggttaacgc agaacctgct aaggttgaca tgaatgtcac 7141 caaacataaa attagttatg ctatttatat gtctggtgga gatgttttgc ttaacgttct 7201 ttcgaagtgt gccggctcta aaaaattcag gcctgctccc gccgctgctt ttgcacgtga 7261 gtgccgcgga ttttatttcg agttacaaga attgaaggaa gacgattatt atgggattac 7321 tttatctgat gattctgatc atcagttttt gcttggatcc caggttgtcg tccatgcatg 7381 cggtggcctg accggtctga actcaggcct cacgacaaat cctggtgtat ccgcttggca 7441 ggtcaacaca gcttatactg cgggacaatt ggtcacatat aacggcaaga cgtataaatg 7501 tttgcagccc cacacctcct tggcaggatg ggaaccatcc aacgttcctg ccttgtggca 7561 gcttcaatga ctgcaggaag gggatccggc tgctaacaaa gcccgaaagg aagctgagtt 7621 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt 7681 gaggggtttt ttgctgaaag gaggaactat atccggat //